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Sample records for multiphoton spectral imaging

  1. Compact multiphoton/single photon laser scanning microscope for spectral imaging and fluorescence lifetime imaging.

    PubMed

    Ulrich, Volker; Fischer, Peter; Riemann, Iris; Königt, Karsten

    2004-01-01

    An inverted fluorescence microscope was upgraded into a compact three-dimensional laser scanning microscope (LSM) of 65 x 62 x 48 cm dimensions by means of a fast kHz galvoscanner unit, a piezodriven z-stage, and a picosecond (ps) 50 MHz laser diode at 405 nm. In addition, compact turn-key near infrared femtosecond lasers have been employed to perform multiphoton fluorescence and second harmonic generation (SHG) microscopy. To expand the features of the compact LSM, a time-correlated single photon counting unit as well as a Sagnac interferometer have been added to realize fluorescence lifetime imaging (FLIM) and spectral imaging. Using this unique five-dimensional microscope, TauMap, single-photon excited (SPE), and two-photon excited (TPE) cellular fluorescence as well as intratissue autofluorescence of water plant leaves have been investigated with submicron spatial resolution, <270 ps temporal resolution, and 10 nm spectral resolution. PMID:15536977

  2. Integrated coherent anti-Stokes Raman scattering and multiphoton microscopy for biomolecular imaging using spectral filtering of a femtosecond laser

    NASA Astrophysics Data System (ADS)

    Lu, Fake; Zheng, Wei; Lin, Jian; Huang, Zhiwei

    2010-03-01

    We report on a unique implementation of a dual 4-f paired-gratings spectral filtering of a femtosecond (fs) laser to realize high contrast coherent anti-Stokes Raman scattering (CARS) microscopy and high quality multiphoton microscopy on the same platform for label-free biomolecular imaging. The 4-f gratings filtering enables readily conversion of the 100 fs laser pulse trains into transform-limited 1 picosecond (ps) pulses in the spectral range of 700-1600 nm for CARS imaging. Compared with fs-CARS microscopy, our ps-CARS imaging yields an approximately tenfold improvement in spectral resolution, while a twofold improvement in nonresonant background suppression. High quality second-harmonic generation and two-photon excitation fluorescence imaging can also be acquired in tandem when swapping the 4-f grating filtering from the ps mode to fs mode. We demonstrate this multimodal nonlinear optical microscopy technique by imaging 465 nm polystyrene beads in water and animal liver tissue.

  3. Quantitative multiphoton imaging

    NASA Astrophysics Data System (ADS)

    König, Karsten; Weinigel, Martin; Breunig, Hans Georg; Uchugonova, Aisada

    2014-02-01

    Certified clinical multiphoton tomographs for label-free multidimensional high-resolution in vivo imaging have been introduced to the market several years ago. Novel tomographs include a flexible 360° scan head attached to a mechanooptical arm for autofluorescence and SHG imaging as well as a CARS module. Non-fluorescent lipids and water, mitochondrial fluorescent NAD(P)H, fluorescent elastin, keratin, and melanin as well as SHG-active collagen can be imaged in vivo with submicron resolution in human skin. Sensitive and rapid detectors allow single photon counting and the construction of 3D maps where the number of detected photons per voxel is depicted. Intratissue concentration profiles from endogenous as well exogenous substances can be generated when the number of detected photons can be correlated with the number of molecules with respect to binding and scattering behavior. Furthermore, the skin ageing index SAAID based on the ratio elastin/collagen as well as the epidermis depth based on the onset of SHG generation can be determined.

  4. Multiphoton Microscopy for Ophthalmic Imaging

    PubMed Central

    Gibson, Emily A.; Masihzadeh, Omid; Lei, Tim C.; Ammar, David A.; Kahook, Malik Y.

    2011-01-01

    We review multiphoton microscopy (MPM) including two-photon autofluorescence (2PAF), second harmonic generation (SHG), third harmonic generation (THG), fluorescence lifetime (FLIM), and coherent anti-Stokes Raman Scattering (CARS) with relevance to clinical applications in ophthalmology. The different imaging modalities are discussed highlighting the particular strength that each has for functional tissue imaging. MPM is compared with current clinical ophthalmological imaging techniques such as reflectance confocal microscopy, optical coherence tomography, and fluorescence imaging. In addition, we discuss the future prospects for MPM in disease detection and clinical monitoring of disease progression, understanding fundamental disease mechanisms, and real-time monitoring of drug delivery. PMID:21274261

  5. Development of a spectrally resolved multifocal multiphoton microscope

    NASA Astrophysics Data System (ADS)

    Liu, Lixin; Shao, Yonghong; Qu, Junle; Li, Heng; Guo, Baoping; Liu, Wenqing; Niu, Hanben

    2008-12-01

    Multifocal Multiphoton Microscopy (MMM) can acquire three-dimensional (3D) fluorescence microscopic images of samples by multiphoton excitation with the advantages of high speed, reduced photobleaching, enhanced penetration depth and high signal-to-noise ratio. As fluorescence spectrum can provide information about the components of the sample, it is becoming increasingly popular in biomedicine to combine fluorescence spectrum measurement with multidimensional fluorescence microscopy. In this paper, we present the development of a spectrally resolved multifocal multiphoton microscope (SR-MMM). A microlens array is employed in the MMM system to produce 2D excitation foci on the sample for simultaneous two-photon excitation and a pair of galvo mirrors is used to scan the excitation foci across the sample. A liquid crystal tunable filter (LCTF) is mounted in front of a high-speed cooled CCD camera and is used to change the detection wavelength of the MMM system. Depth-resolved and spectrum-resolved two-photon excitation fluorescence images of a few samples are obtained with the SR-MMM system.

  6. Analysis of intracellular pH (pHcyt) in mouse models of angiogenesis and carcinogenesis by spectral imaging microscopy, real-time confocal imaging microscopy, and multiphoton spectral imaging

    NASA Astrophysics Data System (ADS)

    Sennoune, Souad; Maiti, Debasish; Martinez-Zaguilan, Raul

    2003-07-01

    We have shown that a specific cytosolic pH (pHcyt) regulatory mechanism, i.e., vacuolar type H+-ATPases at the plasma membrane (pmV-ATPases), allows angiogenic and metastatic cells to survive in an acidic and hostile environment. However, a functional evaluation of this pump's activity in situ (i.e., in living animal models) has not been attempted. We developed a mouse model of angiogenesis and metastasis based on the dorsal skin fold chamber, and implanted highly metastatic human tumor cells that have been engineered to express green fluorescent protein (GFP). GFP can be used as a pH reporter because its fluorescence is pH sensitive. Our studies in isolated single cells indicated that there are distinct pHcyt gradients in the invadipodia versus the lamellipodia due to the preferential expression of pmV-ATPases at the leading edge. We hypothesize that in vivo, these pH gradients also exist. We employed spectral imaging and real time confocal imaging microscopy, since these approaches are complementary and exhibited unsurpassed temporal and spectral resolution, thus allowing us to study pHcyt in discrete subcellular regions of the cells expressing GFP. We can acquire a full frame (i.e., 512 x 512 pixels) in real time confocal imaging at ca. 25-50 msec, whereas spectral imaging allow us to obtain spectral information from discrete domains of ca. 10 μm in the x-y plane and every 10 μm from leading to lagging edge within a time frame of 5 msec at 0.4 nm spectral resolution. This is possible because we employ frame transfer cooled CCD cameras and spectrographs. Studies are under way to evaluate proton gradients using multiphoton approaches since this will allow us to evaluate pH deeper into the tissue (i.e., 300-600 μm), and should allow us to follow pHcyt and the progression of tumor metastasis.

  7. Multiphoton imaging with a nanosecond supercontinuum source

    NASA Astrophysics Data System (ADS)

    Lefort, Claire; O'Connor, Rodney P.; Blanquet, Véronique; Baraige, Fabienne; Tombelaine, Vincent; Lévêque, Philippe; Couderc, Vincent; Leproux, Philippe

    2016-03-01

    Multiphoton microscopy is a well-established technique for biological imaging of several kinds of targets. It is classically based on multiphoton processes allowing two means of contrast simultaneously: two-photon fluorescence (TPF) and second harmonic generation (SHG). Today, the quasi exclusive laser technology used in that aim is femtosecond titanium sapphire (Ti: Sa) laser. We experimentally demonstrate that a nanosecond supercontinuum laser source (STM-250-VIS-IR-custom, Leukos, France; 1 ns, 600-2400 nm, 250 kHz, 1 W) allows to obtain the same kind of image quality in the case of both TPF and SHG, since it is properly filtered. The first set of images concerns the muscle of a mouse. It highlights the simultaneous detection of TPF and SHG. TPF is obtained thanks to the labelling of alpha-actinin with Alexa Fluor® 546 by immunochemistry. SHG is created from the non-centrosymmetric organization of myosin. As expected, discs of actin and myosin are superimposed alternatively. The resulting images are compared with those obtained from a standard femtosecond Ti: Sa source. The physical parameters of the supercontinuum are discussed. Finally, all the interest of using an ultra-broadband source is presented with images obtained in vivo on the brain of a mouse where tumor cells labeled with eGFP are grafted. Texas Red® conjugating Dextran is injected into the blood vessels network. Thus, two fluorophores having absorption wavelengths separated by 80 nm are imaged simultaneously with a single laser source.

  8. Multiphoton microscopy as a diagnostic imaging modality for lung cancer

    NASA Astrophysics Data System (ADS)

    Pavlova, Ina; Hume, Kelly R.; Yazinski, Stephanie A.; Peters, Rachel M.; Weiss, Robert S.; Webb, Watt W.

    2010-02-01

    Lung cancer is the leading killer among all cancers for both men and women in the US, and is associated with one of the lowest 5-year survival rates. Current diagnostic techniques, such as histopathological assessment of tissue obtained by computed tomography guided biopsies, have limited accuracy, especially for small lesions. Early diagnosis of lung cancer can be improved by introducing a real-time, optical guidance method based on the in vivo application of multiphoton microscopy (MPM). In particular, we hypothesize that MPM imaging of living lung tissue based on twophoton excited intrinsic fluorescence and second harmonic generation can provide sufficient morphologic and spectroscopic information to distinguish between normal and diseased lung tissue. Here, we used an experimental approach based on MPM with multichannel fluorescence detection for initial discovery that MPM spectral imaging could differentiate between normal and neoplastic lung in ex vivo samples from a murine model of lung cancer. Current results indicate that MPM imaging can directly distinguish normal and neoplastic lung tissues based on their distinct morphologies and fluorescence emission properties in non-processed lung tissue. Moreover, we found initial indication that MPM imaging differentiates between normal alveolar tissue, inflammatory foci, and lung neoplasms. Our long-term goal is to apply results from ex vivo lung specimens to aid in the development of multiphoton endoscopy for in vivo imaging of lung abnormalities in various animal models, and ultimately for the diagnosis of human lung cancer.

  9. Studies on wide-field-of-view multiphoton imaging using the flexible clinical multiphoton tomograph MPTflex

    NASA Astrophysics Data System (ADS)

    Weinigel, Martin; Breunig, Hans Georg; Fischer, Peter; Kellner-Höfer, Marcel; Bückle, Rainer; König, Karsten

    2012-03-01

    Multiphoton imaging systems are capable of high-resolution 3-D image acquisition of deep tissue. A first commercially available CE-certified biomedical system for subcelluar resolution of human skin has been launched by JenLab company with the DermaInspectR in 2002. The demand for more flexibility caused the development of the MPTflexR, which provides an increased flexibility and accessibility especially for clinical and cosmetic examinations. However the high resolution of clinical multiphoton tomographs are adherent with a small field-of-view (FOV) of about 360×360μm2. Especially time-consuming is the relocation of areas of interest (AOI) like lesions, sweat glands or hair shafts during a multiphoton examination. This limitation can be be overcome by macroscopic large-area (wide-field-ofview) multiphoton tomography, which is tested first within this work.

  10. Multiphoton imaging with high peak power VECSELs

    NASA Astrophysics Data System (ADS)

    Mirkhanov, Shamil; Quarterman, Adrian H.; Swift, Samuel; Praveen, Bavishna B.; Smyth, Conor J. C.; Wilcox, Keith G.

    2016-03-01

    Multiphoton imaging (MMPI) has become one of thee key non-invasive light microscopy techniques. This technique allows deep tissue imaging with high resolution and less photo-damage than conventional confocal microscopy. MPI is type of laser-scanning microscopy that employs localized nonlinear excitation, so that fluorescence is excited only with is scanned focal volume. For many years, Ti: sapphire femtosecond lasers have been the leading light sources for MPI applications. However, recent developments in laser sources and new types of fluorophores indicate that longer wavelength excitation could be a good alternative for these applications. Mode-locked VECSEELs have the potential to be low cost, compact light sources for MPI systems, with the additional advantage of broad wavelength coverage through use of different semiconductor material systems. Here, we use a femtosecond fibber laser to investigate the effect average power and repetition rate has on MPI image quality, to allow us to optimize our mode-locked VVECSELs for MPI.

  11. Cell-based and in vivo spectral analysis of fluorescent proteins for multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Salomonnson, Emma; Mihalko, Laura Anne; Verkhusha, Vladislav V.; Luker, Kathryn E.; Luker, Gary D.

    2012-09-01

    Multiphoton microscopy of cells and subcellular structures labeled with fluorescent proteins is the state-of-the-art technology for longitudinal imaging studies in tissues and living animals. Successful analysis of separate cell populations or signaling events by intravital microscopy requires optimal pairing of multiphoton excitation wavelengths with spectrally distinct fluorescent proteins. While prior studies have analyzed two photon absorption properties of isolated fluorescent proteins, there is limited information about two photon excitation and fluorescence emission profiles of fluorescent proteins expressed in living cells and intact tissues. Multiphoton microscopy was used to analyze fluorescence outputs of multiple blue, green, and red fluorescent proteins in cultured cells and orthotopic tumor xenografts of human breast cancer cells. It is shown that commonly used orange and red fluorescent proteins are excited efficiently by 750 to 760 nm laser light in living cells, enabling dual color imaging studies with blue or cyan proteins without changing excitation wavelength. It is also shown that small incremental changes in excitation wavelength significantly affect emission intensities from fluorescent proteins, which can be used to optimize multi-color imaging using a single laser wavelength. These data will direct optimal selection of fluorescent proteins for multispectral two photon microscopy.

  12. Spectral behavior of second harmonic signals from organic and non-organic materials in multiphoton microscopy

    PubMed Central

    Ehmke, Tobias; Knebl, Andreas; Reiss, Stephan; Fischinger, Isaak R.; Seiler, Theo G.; Stachs, Oliver; Heisterkamp, Alexander

    2015-01-01

    Multimodal nonlinear microscopy allows imaging of highly ordered biological tissue due to spectral separation of nonlinear signals. This requires certain knowledge about the spectral distribution of the different nonlinear signals. In contrast to several publications we demonstrate a factor of 122 relating the full width at half maximum of a gaussian laser pulse spectrum to the corresponding second harmonic pulse spectrum in the spatial domain by using a simple theoretical model. Experiments on monopotassium phosphate crystals (KDP-crystals) and on porcine corneal tissue support our theoretical predictions. Furthermore, no differences in spectral width were found for epi- and trans-detection of the second harmonic signal. Overall, these results may help to build an optimized multiphoton setup for spectral separation of nonlinear signals. PMID:26339527

  13. Spectral behavior of second harmonic signals from organic and non-organic materials in multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Ehmke, Tobias; Knebl, Andreas; Reiss, Stephan; Fischinger, Isaak R.; Seiler, Theo G.; Stachs, Oliver; Heisterkamp, Alexander

    2015-08-01

    Multimodal nonlinear microscopy allows imaging of highly ordered biological tissue due to spectral separation of nonlinear signals. This requires certain knowledge about the spectral distribution of the different nonlinear signals. In contrast to several publications we demonstrate a factor of /1 2 √{ 2 } relating the full width at half maximum of a gaussian laser pulse spectrum to the corresponding second harmonic pulse spectrum in the spatial domain by using a simple theoretical model. Experiments on monopotassium phosphate crystals (KDP-crystals) and on porcine corneal tissue support our theoretical predictions. Furthermore, no differences in spectral width were found for epi- and trans-detection of the second harmonic signal. Overall, these results may help to build an optimized multiphoton setup for spectral separation of nonlinear signals.

  14. Multiphoton imaging of biological samples during freezing and heating

    NASA Astrophysics Data System (ADS)

    Breunig, H. G.; Uchugonova, A.; König, K.

    2014-02-01

    We applied multiphoton microscopic imaging to observe freezing and heating effects in plant- and animal cell samples. The experimental setups consisted of a multiphoton imaging system and a heating and cooling stage which allows for precise temperature control from liquid nitrogen temperature (-196°C 77 K) up to +600°C (873 K) with heating/freezing rates between 0.01 K/min and 150 K/min. Two multiphoton imaging systems were used: a system based on a modified optical microscope and a flexible mobile system. To illustrate the imaging capabilities, plant leafs as well as animal cells were microscopically imaged in vivo during freezing based on autofluorescence lifetime and intensity of intrinsic molecules. The measurements illustrate the usefulness of multiphoton imaging to investigate freezing effects on animal and plant cells.

  15. Intravital multiphoton microscopy for imaging hepatobiliary function

    NASA Astrophysics Data System (ADS)

    Li, Feng-Chieh; Sun, Tzu-Lin; Lee, Hsuan-Shu; Yang, Shu-Mei; Dong, Chen-Yuan

    2007-07-01

    Liver is the chemical factory in body responsible for important functions such as metabolism and detoxification. When liver can not be regenerated in time to amend damages that has occurred, failure of hepatic functions can result. Traditionally, the study of liver pathology has depended on histological techniques, but such methods are limited to ex-vivo observation. In order to study hepatic metabolism in vivo, we have designed a hepatic imaging chamber made of biocompatible titanium alloy (6V4Al-Ti, ELI grade). In combination with multiphoton and second harmonic generation microscopy, our approach allows the intravital observation of hepatic intravital activities to be achieved. Processes such as hepatic metabolism and disease progression can be studied using this methodology.

  16. Widefield multiphoton microscopy with image-based adaptive optics

    NASA Astrophysics Data System (ADS)

    Chang, C.-Y.; Cheng, L.-C.; Su, H.-W.; Yen, W.-C.; Chen, S.-J.

    2012-10-01

    Unlike conventional multiphoton microscopy according to pixel by pixel point scanning, a widefield multiphoton microscope based on spatiotemporal focusing has been developed to provide fast optical sectioning images at a frame rate over 100 Hz. In order to overcome the aberrations of the widefield multiphoton microscope and the wavefront distortion from turbid biospecimens, an image-based adaptive optics system (AOS) was integrated. The feedback control signal of the AOS was acquired according to locally maximize image intensity, which were provided by the widefield multiphoton excited microscope, by using a hill climbing algorithm. Then, the control signal was utilized to drive a deformable mirror in such a way as to eliminate the aberration and distortion. A R6G-doped PMMA thin film is also increased by 3.7-fold. Furthermore, the TPEF image quality of 1 μm fluorescent beads sealed in agarose gel at different depths is improved.

  17. Optical clearing and multiphoton imaging of paraffin-embedded specimens

    NASA Astrophysics Data System (ADS)

    Wilson, Jesse W.; Degan, Simone; Fischer, Martin C.; Warren, Warren S.

    2013-02-01

    New labeling, imaging, or analysis tools could provide new retrospective insights when applied to archived, paraffin-embedded samples. Deep-tissue multiphoton microscopy of paraffin-embedded specimens is achieved using optical clearing with mineral oil. We tested a variety of murine tissue specimens including skin, lung, spleen, kidney, and heart, acquiring multiphoton autofluorescence and second-harmonic generation, and pump-probe images This technique introduces the capability for non-destructive 3-dimensional microscopic imaging of existing archived pathology specimens, enabling retrospective studies.

  18. Coherence-Gated Sensorless Adaptive Optics Multiphoton Retinal Imaging.

    PubMed

    Cua, Michelle; Wahl, Daniel J; Zhao, Yuan; Lee, Sujin; Bonora, Stefano; Zawadzki, Robert J; Jian, Yifan; Sarunic, Marinko V

    2016-09-07

    Multiphoton microscopy enables imaging deep into scattering tissues. The efficient generation of non-linear optical effects is related to both the pulse duration (typically on the order of femtoseconds) and the size of the focused spot. Aberrations introduced by refractive index inhomogeneity in the sample distort the wavefront and enlarge the focal spot, which reduces the multiphoton signal. Traditional approaches to adaptive optics wavefront correction are not effective in thick or multi-layered scattering media. In this report, we present sensorless adaptive optics (SAO) using low-coherence interferometric detection of the excitation light for depth-resolved aberration correction of two-photon excited fluorescence (TPEF) in biological tissue. We demonstrate coherence-gated SAO TPEF using a transmissive multi-actuator adaptive lens for in vivo imaging in a mouse retina. This configuration has significant potential for reducing the laser power required for adaptive optics multiphoton imaging, and for facilitating integration with existing systems.

  19. Coherence-Gated Sensorless Adaptive Optics Multiphoton Retinal Imaging

    NASA Astrophysics Data System (ADS)

    Cua, Michelle; Wahl, Daniel J.; Zhao, Yuan; Lee, Sujin; Bonora, Stefano; Zawadzki, Robert J.; Jian, Yifan; Sarunic, Marinko V.

    2016-09-01

    Multiphoton microscopy enables imaging deep into scattering tissues. The efficient generation of non-linear optical effects is related to both the pulse duration (typically on the order of femtoseconds) and the size of the focused spot. Aberrations introduced by refractive index inhomogeneity in the sample distort the wavefront and enlarge the focal spot, which reduces the multiphoton signal. Traditional approaches to adaptive optics wavefront correction are not effective in thick or multi-layered scattering media. In this report, we present sensorless adaptive optics (SAO) using low-coherence interferometric detection of the excitation light for depth-resolved aberration correction of two-photon excited fluorescence (TPEF) in biological tissue. We demonstrate coherence-gated SAO TPEF using a transmissive multi-actuator adaptive lens for in vivo imaging in a mouse retina. This configuration has significant potential for reducing the laser power required for adaptive optics multiphoton imaging, and for facilitating integration with existing systems.

  20. Coherence-Gated Sensorless Adaptive Optics Multiphoton Retinal Imaging.

    PubMed

    Cua, Michelle; Wahl, Daniel J; Zhao, Yuan; Lee, Sujin; Bonora, Stefano; Zawadzki, Robert J; Jian, Yifan; Sarunic, Marinko V

    2016-01-01

    Multiphoton microscopy enables imaging deep into scattering tissues. The efficient generation of non-linear optical effects is related to both the pulse duration (typically on the order of femtoseconds) and the size of the focused spot. Aberrations introduced by refractive index inhomogeneity in the sample distort the wavefront and enlarge the focal spot, which reduces the multiphoton signal. Traditional approaches to adaptive optics wavefront correction are not effective in thick or multi-layered scattering media. In this report, we present sensorless adaptive optics (SAO) using low-coherence interferometric detection of the excitation light for depth-resolved aberration correction of two-photon excited fluorescence (TPEF) in biological tissue. We demonstrate coherence-gated SAO TPEF using a transmissive multi-actuator adaptive lens for in vivo imaging in a mouse retina. This configuration has significant potential for reducing the laser power required for adaptive optics multiphoton imaging, and for facilitating integration with existing systems. PMID:27599635

  1. Coherence-Gated Sensorless Adaptive Optics Multiphoton Retinal Imaging

    PubMed Central

    Cua, Michelle; Wahl, Daniel J.; Zhao, Yuan; Lee, Sujin; Bonora, Stefano; Zawadzki, Robert J.; Jian, Yifan; Sarunic, Marinko V.

    2016-01-01

    Multiphoton microscopy enables imaging deep into scattering tissues. The efficient generation of non-linear optical effects is related to both the pulse duration (typically on the order of femtoseconds) and the size of the focused spot. Aberrations introduced by refractive index inhomogeneity in the sample distort the wavefront and enlarge the focal spot, which reduces the multiphoton signal. Traditional approaches to adaptive optics wavefront correction are not effective in thick or multi-layered scattering media. In this report, we present sensorless adaptive optics (SAO) using low-coherence interferometric detection of the excitation light for depth-resolved aberration correction of two-photon excited fluorescence (TPEF) in biological tissue. We demonstrate coherence-gated SAO TPEF using a transmissive multi-actuator adaptive lens for in vivo imaging in a mouse retina. This configuration has significant potential for reducing the laser power required for adaptive optics multiphoton imaging, and for facilitating integration with existing systems. PMID:27599635

  2. In vivo multiphoton imaging of obstructive cholestasis in mice

    NASA Astrophysics Data System (ADS)

    Li, Feng-Chieh; Lee, Yu Yang; Chiou, Ling-Ling; Lee, Hsuan-Shu; Dong, Chen-Yuan

    2010-02-01

    Combining multiphoton microscopy with a newly designed hepatic imaging window, we acquired in vivo images of mice obstructive cholestasis. We observed that in mice with bile duct ligation, bile canaliculi failed to appear during the whole observation period over 100 minutes following carboxyfluorescein diacetate injection, whereas the fluorescence was retained much longer within sinusoids. Furthermore, the fluorescence intensities in sinusoids were persistently higher than in hepatocytes during the course.

  3. Moxifloxacin: Clinically compatible contrast agent for multiphoton imaging

    PubMed Central

    Wang, Taejun; Jang, Won Hyuk; Lee, Seunghun; Yoon, Calvin J.; Lee, Jun Ho; Kim, Bumju; Hwang, Sekyu; Hong, Chun-Pyo; Yoon, Yeoreum; Lee, Gilgu; Le, Viet-Hoan; Bok, Seoyeon; Ahn, G-One; Lee, Jaewook; Gho, Yong Song; Chung, Euiheon; Kim, Sungjee; Jang, Myoung Ho; Myung, Seung-Jae; Kim, Myoung Joon; So, Peter T. C.; Kim, Ki Hean

    2016-01-01

    Multiphoton microscopy (MPM) is a nonlinear fluorescence microscopic technique widely used for cellular imaging of thick tissues and live animals in biological studies. However, MPM application to human tissues is limited by weak endogenous fluorescence in tissue and cytotoxicity of exogenous probes. Herein, we describe the applications of moxifloxacin, an FDA-approved antibiotic, as a cell-labeling agent for MPM. Moxifloxacin has bright intrinsic multiphoton fluorescence, good tissue penetration and high intracellular concentration. MPM with moxifloxacin was demonstrated in various cell lines, and animal tissues of cornea, skin, small intestine and bladder. Clinical application is promising since imaging based on moxifloxacin labeling could be 10 times faster than imaging based on endogenous fluorescence. PMID:27283889

  4. Moxifloxacin: Clinically compatible contrast agent for multiphoton imaging

    NASA Astrophysics Data System (ADS)

    Wang, Taejun; Jang, Won Hyuk; Lee, Seunghun; Yoon, Calvin J.; Lee, Jun Ho; Kim, Bumju; Hwang, Sekyu; Hong, Chun-Pyo; Yoon, Yeoreum; Lee, Gilgu; Le, Viet-Hoan; Bok, Seoyeon; Ahn, G.-One; Lee, Jaewook; Gho, Yong Song; Chung, Euiheon; Kim, Sungjee; Jang, Myoung Ho; Myung, Seung-Jae; Kim, Myoung Joon; So, Peter T. C.; Kim, Ki Hean

    2016-06-01

    Multiphoton microscopy (MPM) is a nonlinear fluorescence microscopic technique widely used for cellular imaging of thick tissues and live animals in biological studies. However, MPM application to human tissues is limited by weak endogenous fluorescence in tissue and cytotoxicity of exogenous probes. Herein, we describe the applications of moxifloxacin, an FDA-approved antibiotic, as a cell-labeling agent for MPM. Moxifloxacin has bright intrinsic multiphoton fluorescence, good tissue penetration and high intracellular concentration. MPM with moxifloxacin was demonstrated in various cell lines, and animal tissues of cornea, skin, small intestine and bladder. Clinical application is promising since imaging based on moxifloxacin labeling could be 10 times faster than imaging based on endogenous fluorescence.

  5. Microstructure imaging of human rectal mucosa using multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Liu, N. R.; Chen, G.; Chen, J. X.; Yan, J.; Zhuo, S. M.; Zheng, L. Q.; Jiang, X. S.

    2011-01-01

    Multiphoton microscopy (MPM) has high resolution and sensitivity. In this study, MPM was used to image microstructure of human rectal mucosa. The morphology and distribution of the main components in mucosa layer, absorptive cells and goblet cells in the epithelium, abundant intestinal glands in the lamina propria and smooth muscle fibers in the muscularis mucosa were clearly monitored. The variations of these components were tightly relevant to the pathology in gastrointestine system, especially early rectal cancer. The obtained images will be helpful for the diagnosis of early colorectal cancer.

  6. In vivo multiphoton imaging of bile duct ligation

    NASA Astrophysics Data System (ADS)

    Liu, Yuan; Li, Feng-Chieh; Chen, Hsiao-Chin; Chang, Po-shou; Yang, Shu-Mei; Lee, Hsuan-Shu; Dong, Chen-Yuan

    2008-02-01

    Bile is the exocrine secretion of liver and synthesized by hepatocytes. It is drained into duodenum for the function of digestion or drained into gallbladder for of storage. Bile duct obstruction is a blockage in the tubes that carry bile to the gallbladder and small intestine. However, Bile duct ligation results in the changes of bile acids in serum, liver, urine, and feces1, 2. In this work, we demonstrate a novel technique to image this pathological condition by using a newly developed in vivo imaging system, which includes multiphoton microscopy and intravital hepatic imaging chamber. The images we acquired demonstrate the uptake, processing of 6-CFDA in hepatocytes and excretion of CF in the bile canaliculi. In addition to imaging, we can also measure kinetics of the green fluorescence intensity.

  7. Multiphoton imaging: a view to understanding sulfur mustard lesions

    NASA Astrophysics Data System (ADS)

    Werrlein, Robert J. S.; Madren-Whalley, Janna S.

    2003-07-01

    It is well known that topical exposure to sulfur mustard (SM) produces persistent, incapacitating blisters of the skin. However, the primary lesions effecting epidermal-dermal separation and disabling of mechanisms for cutaneous repair remain uncertain. Immunofluorescent staining plus multiphoton imaging of human epidermal tissues and keratinocytes exposed to SM (400 μM x 5 min)have revealed that SM disrupts adhesion-complex molecules which are also disrupted by epidermolysis bullosa-type blistering diseases of the skin. Images of keratin-14 showed early, progressive, postexposure collapse of the K5/K14 cytoskeleton that resulted in ventral displacement of the nuclei beneath its collapsing filaments. This effectively corrupted the dynamic filament assemblies that link basal-cell nuclei to the extracellular matrix via α6β4-integrin and laminin-5. At 1 h postexposure, there was disruption in the surface organization of α6β4 integrins, associated displacement of laminin-5 anchoring sites and a concomitant loss of functional asymmetry. Accordingly, our multiphoton images are providing compelling evidence that SM induces prevesicating lesions that disrupt the receptor-ligand organization and cytoskeletal systems required for maintaining dermal-epidermal attachment, signal transduction, and polarized mobility.

  8. Molecule-specific darkfield and multiphoton imaging using gold nanocages

    NASA Astrophysics Data System (ADS)

    Powless, Amy J.; Jenkins, Samir V.; McKay, Mary Lee; Chen, Jingyi; Muldoon, Timothy J.

    2015-03-01

    Due to their robust optical properties, biological inertness, and readily adjustable surface chemistry, gold nanostructures have been demonstrated as contrast agents in a variety of biomedical imaging applications. One application is dynamic imaging of live cells using bioconjugated gold nanoparticles to monitor molecule trafficking mechanisms within cells; for instance, the regulatory pathway of epidermal growth factor receptor (EGFR) undergoing endocytosis. In this paper, we have demonstrated a method to track endocytosis of EGFR in MDA-MB-468 breast adenocarcinoma cells using bioconjugated gold nanocages (AuNCs) and multiphoton microscopy. Dynamic imaging was performed using a time series capture of 4 images every minute for one hour. Specific binding and internalization of the bioconjugated AuNCs was observed while the two control groups showed non-specific binding at fewer surface sites, leading to fewer bound AuNCs and no internalization.

  9. Multifunctional gold nanorod theragnostics probed by multi-photon imaging.

    PubMed

    Book Newell, Brittany; Wang, Yuling; Irudayaraj, Joseph

    2012-02-01

    This study exhibits the fabrication of target-specific Gold nanorods (GNRs) coupled with an anti-tumorigenic apoptotic drug and provides tracking of the labeled particles as they migrate through cells and release their drug-load to targeted cancer cells. We utilize the photoluminescence property of GNRs and their ability to be conjugated with multiple agents to transform facile rods to a targeted drug delivery vehicle. GNRs of aspect ratio 2.8 were conjugated with a targeting ligand, folic acid and an anthracycline drug, Doxorubicin. The multifunctional nanorods were then used to target folate receptor expressing cancers cells for the delivery of a concentration dependent dosage of Doxorubicin (DOX). By utilizing the photoluminescence of GNRs and the innate fluorescence of DOX, multi-photon fluorescence lifetime imaging was utilized to monitor the uptake of functionalized nanorods, the release of the drug and its localization in living cells. We show that these nano-vehicles successfully targeted cancer cells over expressing folate receptors and showed low toxicity to control cell lines. Release of DOX was observed in the cytoplasmic region and after 16 h was found to be redistributed in the nucleus resulting in cell death. Our theragnostic approach demonstrates the fabrication of multifunctional GNRs for targeted drug delivery and monitoring of the drug and the vehicle by multi-photon microscopy using fluorescence intensity and lifetime imaging.

  10. Live-Animal Imaging of Renal Function by Multiphoton Microscopy

    PubMed Central

    Dunn, Kenneth W.; Sutton, Timothy A.; Sandoval, Ruben M.

    2015-01-01

    Intravital microscopy, microscopy of living animals, is a powerful research technique that combines the resolution and sensitivity found in microscopic studies of cultured cells with the relevance and systemic influences of cells in the context of the intact animal. The power of intravital microscopy has recently been extended with the development of multiphoton fluorescence microscopy systems capable of collecting optical sections from deep within the kidney at subcellular resolution, supporting high-resolution characterizations of the structure and function of glomeruli, tubules, and vasculature in the living kidney. Fluorescent probes are administered to an anesthetized, surgically prepared animal, followed by image acquisition for up to 3 hr. Images are transferred via a high-speed network to specialized computer systems for digital image analysis. This general approach can be used with different combinations of fluorescent probes to evaluate processes such as glomerular permeability, proximal tubule endocytosis, microvascular flow, vascular permeability, mitochondrial function, and cellular apoptosis/necrosis. PMID:23042524

  11. Multiphoton fluorescence excitation: new spectral windows for biological nonlinear microscopy.

    PubMed Central

    Xu, C; Zipfel, W; Shear, J B; Williams, R M; Webb, W W

    1996-01-01

    Intrinsic, three-dimensionally resolved, microscopic imaging of dynamical structures and biochemical processes in living preparations has been realized by nonlinear laser scanning fluorescence microscopy. The search for useful two-photon and three-photon excitation spectra, motivated by the emergence of nonlinear microscopy as a powerful biophysical instrument, has now discovered a virtual artist's palette of chemical indicators, fluorescent markers, and native biological fluorophores, including NADH, flavins, and green fluorescent proteins, that are applicable to living biological preparations. More than 25 two-photon excitation spectra of ultraviolet and visible absorbing molecules reveal useful cross sections, some conveniently blue-shifted, for near-infrared absorption. Measurements of three-photon fluorophore excitation spectra now define alternative windows at relatively benign wavelengths to excite deeper ultraviolet fluorophores. The inherent optical sectioning capability of nonlinear excitation provides three-dimensional resolution for imaging and avoids out-of-focus background and photodamage. Here, the measured nonlinear excitation spectra and their photophysical characteristics that empower nonlinear laser microscopy for biological imaging are described. Images Fig. 1 Fig. 5 Fig. 7 PMID:8855254

  12. Achieving molecular selectivity in imaging using multiphoton Raman spectroscopy techniques

    SciTech Connect

    Holtom, Gary R. ); Thrall, Brian D. ); Chin, Beek Yoke ); Wiley, H Steven ); Colson, Steven D. )

    2000-12-01

    In the case of most imaging methods, contrast is generated either by physical properties of the sample (Differential Image Contrast, Phase Contrast), or by fluorescent labels that are localized to a particular protein or organelle. Standard Raman and infrared methods for obtaining images are based upon the intrinsic vibrational properties of molecules, and thus obviate the need for attached flurophores. Unfortunately, they have significant limitations for live-cell imaging. However, an active Raman method, called Coherent Anti-Stokes Raman Scattering (CARS), is well suited for microscopy, and provides a new means for imaging specific molecules. Vibrational imaging techniques, such as CARS, avoid problems associated with photobleaching and photo-induced toxicity often associated with the use of fluorescent labels with live cells. Because the laser configuration needed to implement CARS technology is similar to that used in other multiphoton microscopy methods, such as two -photon fluorescence and harmonic generation, it is possible to combine imaging modalities, thus generating simultaneous CARS and fluorescence images. A particularly powerful aspect of CARS microscopy is its ability to selectively image deuterated compounds, thus allowing the visualization of molecules, such as lipids, that are chemically indistinguishable from the native species.

  13. Direct trabecular meshwork imaging in porcine eyes through multiphoton gonioscopy

    NASA Astrophysics Data System (ADS)

    Masihzadeh, Omid; Ammar, David A.; Kahook, Malik Y.; Gibson, Emily A.; Lei, Tim C.

    2013-03-01

    The development of technologies to characterize the ocular aqueous outflow system (AOS) is important for the understanding of the pathophysiology of glaucoma. Multiphoton microscopy (MPM) offers the advantage of high-resolution, label-free imaging with intrinsic image contrast because the emitted signals result from the specific biomolecular content of the tissue. Previous attempts to use MPM to image the murine irido-corneal region directly through the sclera have suffered from degradation in image resolution due to scattering of the focused laser light. As a result, transscleral MPM has limited ability to observe fine structures in the AOS. In this work, the porcine irido-corneal angle was successfully imaged through the transparent cornea using a gonioscopic lens to circumvent the highly scattering scleral tissue. The resulting high-resolution images allowed the detailed structures in the trabecular meshwork (TM) to be observed. Multimodal imaging by two-photon autofluorescence and second harmonic generation allowed visualization of different features in the TM without labels and without disruption of the TM or surrounding tissues. MPM gonioscopy is a promising noninvasive imaging tool for high-resolution studies of the AOS, and research continues to explore the potential for future clinical applications in humans.

  14. Multiphoton gonioscopy to image the trabecular meshwork of porcine eyes

    NASA Astrophysics Data System (ADS)

    Masihzadeh, Omid; Ammar, David A.; Kahook, Malik Y.; Gibson, Emily A.; Lei, Tim C.

    2013-03-01

    The aqueous outflow system (AOS), including the trabecular meshwork (TM), the collector channels (CC) and the Schlemm's canal (SC), regulates intraocular pressure (IOP) through the drainage of the aqueous humor (AH). Abnormal IOP elevation leads to increased pressure stress to retinal ganglion cells, resulting in cell loss that can ultimately lead to complete loss of eyesight. Therefore, development of imaging tools to detect abnormal structural and functional changes of the AOS is important in early diagnosis and prevention of glaucoma. Multiphoton microscopy (MPM), including twophoton autofluorescence (TPAF) and second harmonic generation (SHG), is a label-free microscopic technique that allows molecular specific imaging of biological tissues like the TM. Since the TM and other AOS structures are located behind the highly scattering scleral tissue, transscleral imaging of the TM does not provide enough optical resolution. In this work, a gonioscopic lens is used to allow direct optical access of the TM through the cornea for MPM imaging. Compared to transscleral imaging, the acquired MPM images show improved resolution as individual collagen fiber bundles of the TM can be observed. MPM gonioscopy may have the potential to be developed as a future clinical imaging tool for glaucoma diagnostics.

  15. High-Resolution Multiphoton Imaging of Tumors In Vivo

    PubMed Central

    Wyckoff, Jeffrey; Gligorijevic, Bojana; Entenberg, David; Segall, Jeffrey; Condeelis, John

    2014-01-01

    Analysis of the individual steps in metastasis is crucial if insights at the molecular level are to be linked to the cell biology of cancer. A technical hurdle to achieving the analysis of the individual steps of metastasis is the fact that, at the gross level, tumors are heterogeneous in both animal models and patients. Human primary tumors show extensive variation in all properties ranging from growth and morphology of the tumor through tumor-cell density in the blood and formation and growth of metastases. Methods capable of the direct visualization and analysis of tumor-cell behavior at single-cell resolution in vivo have become crucial in advancing the understanding of mechanisms of metastasis, the definition of microenvironment, and the markers related to both. This article discusses the use of high-resolution multiphoton imaging of tumors (specifically breast tumors in mice) in vivo. PMID:21969629

  16. High-resolution multiphoton imaging of tumors in vivo.

    PubMed

    Wyckoff, Jeffrey; Gligorijevic, Bojana; Entenberg, David; Segall, Jeffrey; Condeelis, John

    2011-10-01

    Analysis of the individual steps in metastasis is crucial if insights at the molecular level are to be linked to the cell biology of cancer. A technical hurdle to achieving the analysis of the individual steps of metastasis is the fact that, at the gross level, tumors are heterogeneous in both animal models and patients. Human primary tumors show extensive variation in all properties ranging from growth and morphology of the tumor through tumor-cell density in the blood and formation and growth of metastases. Methods capable of the direct visualization and analysis of tumor-cell behavior at single-cell resolution in vivo have become crucial in advancing the understanding of mechanisms of metastasis, the definition of microenvironment, and the markers related to both. This article discusses the use of high-resolution multiphoton imaging of tumors (specifically breast tumors in mice) in vivo.

  17. Superresolved multiphoton microscopy with spatial frequency-modulated imaging.

    PubMed

    Field, Jeffrey J; Wernsing, Keith A; Domingue, Scott R; Allende Motz, Alyssa M; DeLuca, Keith F; Levi, Dean H; DeLuca, Jennifer G; Young, Michael D; Squier, Jeff A; Bartels, Randy A

    2016-06-14

    Superresolved far-field microscopy has emerged as a powerful tool for investigating the structure of objects with resolution well below the diffraction limit of light. Nearly all superresolution imaging techniques reported to date rely on real energy states of fluorescent molecules to circumvent the diffraction limit, preventing superresolved imaging with contrast mechanisms that occur via virtual energy states, including harmonic generation (HG). We report a superresolution technique based on spatial frequency-modulated imaging (SPIFI) that permits superresolved nonlinear microscopy with any contrast mechanism and with single-pixel detection. We show multimodal superresolved images with two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) from biological and inorganic media. Multiphoton SPIFI (MP-SPIFI) provides spatial resolution up to 2η below the diffraction limit, where η is the highest power of the nonlinear intensity response. MP-SPIFI can be used to provide enhanced resolution in optically thin media and may provide a solution for superresolved imaging deep in scattering media. PMID:27231219

  18. Multiphoton and photothermal imaging of molecular events in cancer

    NASA Astrophysics Data System (ADS)

    Skala, Melissa

    2010-10-01

    Optical techniques are attractive for monitoring disease processes in living tissues because they are relatively cheap, non-invasive and provide a wealth of functional information. Multiphoton microscopy (MPM) and Optical Coherence Tomography (OCT) are two types of three-dimensional optical imaging modalities that have demonstrated great utility in pre-clinical models of disease. These techniques are particularly useful for identifying metabolic and molecular biomarkers in cancer. These biomarkers can be used to identify the mechanisms of tumor growth, and to predict the response of a particular tumor to treatment. Specifically, MPM of the co-enzymes NADH and FAD was used to quantify metabolic changes associated with developing cancers in vivo. This imaging technique exploits intrinsic sources of tissue contrast and thus does not require contrast agents. Ongoing work combines this metabolic imaging technique with vascular imaging to provide a comprehensive picture of oxygen supply and demand with tumor therapy. Molecular signaling represents a third critical component in tumor physiology. To this end we have recently developed photothermal OCT, which combines coherent detection with laser-heated gold nanoparticles to achieve high-resolution molecular contrast at deeper depths than MPM. This multi-functional imaging platform will provide unprecedented insight into oxygen supply and demand, and molecular signaling in response to tumor growth and targeted cancer therapies in pre-clinical models.

  19. Water-soluble quantum dots for multiphoton fluorescence imaging in vivo.

    PubMed

    Larson, Daniel R; Zipfel, Warren R; Williams, Rebecca M; Clark, Stephen W; Bruchez, Marcel P; Wise, Frank W; Webb, Watt W

    2003-05-30

    The use of semiconductor nanocrystals (quantum dots) as fluorescent labels for multiphoton microscopy enables multicolor imaging in demanding biological environments such as living tissue. We characterized water-soluble cadmium selenide-zinc sulfide quantum dots for multiphoton imaging in live animals. These fluorescent probes have two-photon action cross sections as high as 47,000 Goeppert-Mayer units, by far the largest of any label used in multiphoton microscopy. We visualized quantum dots dynamically through the skin of living mice, in capillaries hundreds of micrometers deep. We found no evidence of blinking (fluorescence intermittency) in solution on nanosecond to millisecond time scales.

  20. Water-Soluble Quantum Dots for Multiphoton Fluorescence Imaging in Vivo

    NASA Astrophysics Data System (ADS)

    Larson, Daniel R.; Zipfel, Warren R.; Williams, Rebecca M.; Clark, Stephen W.; Bruchez, Marcel P.; Wise, Frank W.; Webb, Watt W.

    2003-05-01

    The use of semiconductor nanocrystals (quantum dots) as fluorescent labels for multiphoton microscopy enables multicolor imaging in demanding biological environments such as living tissue. We characterized water-soluble cadmium selenide-zinc sulfide quantum dots for multiphoton imaging in live animals. These fluorescent probes have two-photon action cross sections as high as 47,000 Goeppert-Mayer units, by far the largest of any label used in multiphoton microscopy. We visualized quantum dots dynamically through the skin of living mice, in capillaries hundreds of micrometers deep. We found no evidence of blinking (fluorescence intermittency) in solution on nanosecond to millisecond time scales.

  1. Water-soluble quantum dots for multiphoton fluorescence imaging in vivo.

    PubMed

    Larson, Daniel R; Zipfel, Warren R; Williams, Rebecca M; Clark, Stephen W; Bruchez, Marcel P; Wise, Frank W; Webb, Watt W

    2003-05-30

    The use of semiconductor nanocrystals (quantum dots) as fluorescent labels for multiphoton microscopy enables multicolor imaging in demanding biological environments such as living tissue. We characterized water-soluble cadmium selenide-zinc sulfide quantum dots for multiphoton imaging in live animals. These fluorescent probes have two-photon action cross sections as high as 47,000 Goeppert-Mayer units, by far the largest of any label used in multiphoton microscopy. We visualized quantum dots dynamically through the skin of living mice, in capillaries hundreds of micrometers deep. We found no evidence of blinking (fluorescence intermittency) in solution on nanosecond to millisecond time scales. PMID:12775841

  2. Multi Spectral Imaging System

    NASA Technical Reports Server (NTRS)

    Spiering, Bruce A. (Inventor)

    1999-01-01

    An optical imaging system provides automatic co-registration of a plurality of multi spectral images of an object which are generated by a plurality of video cameras or other optical detectors. The imaging system includes a modular assembly of beam splitters, lens tubes, camera lenses and wavelength selective filters which facilitate easy reconfiguration and adjustment of the system for various applications. A primary lens assembly generates a real image of an object to be imaged on a reticle which is positioned at a fixed length from a beam splitter assembly. The beam splitter assembly separates a collimated image beam received from the reticle into multiple image beams, each of which is projected onto a corresponding one of a plurality of video cameras. The lens tubes which connect the beam splitter assembly to the cameras are adjustable in length to provide automatic co-registration of the images generated by each camera.

  3. Multipurpose spectral imager.

    PubMed

    Sigernes, F; Lorentzen, D A; Heia, K; Svenøe, T

    2000-06-20

    A small spectral imaging system is presented that images static or moving objects simultaneously as a function of wavelength. The main physical principle is outlined and demonstrated. The instrument is capable of resolving both spectral and spatial information from targets throughout the entire visible region. The spectral domain has a bandpass of 12 A. One can achieve the spatial domain by rotating the system's front mirror with a high-resolution stepper motor. The spatial resolution range from millimeters to several meters depends mainly on the front optics used and whether the target is fixed (static) or movable relative to the instrument. Different applications and examples are explored, including outdoor landscapes, industrial fish-related targets, and ground-level objects observed in the more traditional way from an airborne carrier (remote sensing). Through the examples, we found that the instrument correctly classifies whether a shrimp is peeled and whether it can disclose the spectral and spatial microcharacteristics of targets such as a fish nematode (parasite). In the macroregime, we were able to distinguish a marine vessel from the surrounding sea and sky. A study of the directional spectral albedo from clouds, mountains, snow cover, and vegetation has also been included. With the airborne experiment, the imager successfully classified snow cover, leads, and new and rafted ice, as seen from 10.000 ft (3.048 m). PMID:18345245

  4. Multipurpose spectral imager.

    PubMed

    Sigernes, F; Lorentzen, D A; Heia, K; Svenøe, T

    2000-06-20

    A small spectral imaging system is presented that images static or moving objects simultaneously as a function of wavelength. The main physical principle is outlined and demonstrated. The instrument is capable of resolving both spectral and spatial information from targets throughout the entire visible region. The spectral domain has a bandpass of 12 A. One can achieve the spatial domain by rotating the system's front mirror with a high-resolution stepper motor. The spatial resolution range from millimeters to several meters depends mainly on the front optics used and whether the target is fixed (static) or movable relative to the instrument. Different applications and examples are explored, including outdoor landscapes, industrial fish-related targets, and ground-level objects observed in the more traditional way from an airborne carrier (remote sensing). Through the examples, we found that the instrument correctly classifies whether a shrimp is peeled and whether it can disclose the spectral and spatial microcharacteristics of targets such as a fish nematode (parasite). In the macroregime, we were able to distinguish a marine vessel from the surrounding sea and sky. A study of the directional spectral albedo from clouds, mountains, snow cover, and vegetation has also been included. With the airborne experiment, the imager successfully classified snow cover, leads, and new and rafted ice, as seen from 10.000 ft (3.048 m).

  5. In vivo multiphoton microscopy associated to 3D image processing for human skin characterization

    NASA Astrophysics Data System (ADS)

    Baldeweck, T.; Tancrède, E.; Dokladal, P.; Koudoro, S.; Morard, V.; Meyer, F.; Decencière, E.; Pena, A.-M.

    2012-03-01

    Multiphoton microscopy has emerged in the past decade as a promising non-invasive skin imaging technique. The aim of this study was to assess whether multiphoton microscopy coupled to specific 3D image processing tools could provide new insights into the organization of different skin components and their age-related changes. For that purpose, we performed a clinical trial on 15 young and 15 aged human female volunteers on the ventral and dorsal side of the forearm using the DermaInspectR medical imaging device. We visualized the skin by taking advantage of intrinsic multiphoton signals from cells, elastic and collagen fibers. We also developed 3D image processing algorithms adapted to in vivo multiphoton images of human skin in order to extract quantitative parameters in each layer of the skin (epidermis and superficial dermis). The results show that in vivo multiphoton microscopy is able to evidence several skin alterations due to skin aging: morphological changes in the epidermis and modifications in the quantity and organization of the collagen and elastic fibers network. In conclusion, the association of multiphoton microscopy with specific image processing allows the three-dimensional organization of skin components to be visualized and quantified thus providing a powerful tool for cosmetic and dermatological investigations.

  6. Multiphoton imaging the disruptive nature of sulfur mustard lesions

    NASA Astrophysics Data System (ADS)

    Werrlein, Robert J.; Braue, Catherine R.; Dillman, James F.

    2005-03-01

    Sulfur mustard [bis-2-chloroethyl sulfide] is a vesicating agent first used as a weapon of war in WWI. It causes debilitating blisters at the epidermal-dermal junction and involves molecules that are also disrupted by junctional epidermolysis bullosa (JEB) and other blistering skin diseases. Despite its recurring use in global conflicts, there is still no completely effective treatment. We have shown by imaging human keratinocytes in cell culture and in intact epidermal tissues that the basal cells of skin contain well-organized molecules (keratins K5/K14, α6β4 integrin, laminin 5 and α3β1 integrin) that are early targets of sulfur mustard. Disruption and collapse of these molecules is coincident with nuclear displacement, loss of functional asymmetry, and loss of polarized mobility. The progression of this pathology precedes basal cell detachment by 8-24 h, a time equivalent to the "clinical latent phase" that defines the extant period between agent exposure and vesication. Our images indicate that disruption of adhesion-complex molecules also impairs cytoskeletal proteins and the integration of structures required for signal transduction and tissue repair. We have recently developed an optical system to test this hypothesis, i.e., to determine whether and how the early disruption of target molecules alters signal transduction. This environmentally controlled on-line system provides a nexus for real-time correlation of imaged lesions with DNA microarray analysis, and for using multiphoton microscopy to facilitate development of more effective treatment strategies.

  7. Invited Review Article: Imaging techniques for harmonic and multiphoton absorption fluorescence microscopy

    PubMed Central

    Carriles, Ramón; Schafer, Dawn N.; Sheetz, Kraig E.; Field, Jeffrey J.; Cisek, Richard; Barzda, Virginijus; Sylvester, Anne W.; Squier, Jeffrey A.

    2009-01-01

    We review the current state of multiphoton microscopy. In particular, the requirements and limitations associated with high-speed multiphoton imaging are considered. A description of the different scanning technologies such as line scan, multifoci approaches, multidepth microscopy, and novel detection techniques is given. The main nonlinear optical contrast mechanisms employed in microscopy are reviewed, namely, multiphoton excitation fluorescence, second harmonic generation, and third harmonic generation. Techniques for optimizing these nonlinear mechanisms through a careful measurement of the spatial and temporal characteristics of the focal volume are discussed, and a brief summary of photobleaching effects is provided. Finally, we consider three new applications of multiphoton microscopy: nonlinear imaging in microfluidics as applied to chemical analysis and the use of two-photon absorption and self-phase modulation as contrast mechanisms applied to imaging problems in the medical sciences. PMID:19725639

  8. Optimal spectral filtering in soliton self-frequency shift for deep-tissue multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Wang, Ke; Qiu, Ping

    2015-05-01

    Tunable optical solitons generated by soliton self-frequency shift (SSFS) have become valuable tools for multiphoton microscopy (MPM). Recent progress in MPM using 1700 nm excitation enabled visualizing subcortical structures in mouse brain in vivo for the first time. Such an excitation source can be readily obtained by SSFS in a large effective-mode-area photonic crystal rod with a 1550-nm fiber femtosecond laser. A longpass filter was typically used to isolate the soliton from the residual in order to avoid excessive energy deposit on the sample, which ultimately leads to optical damage. However, since the soliton was not cleanly separated from the residual, the criterion for choosing the optimal filtering wavelength is lacking. Here, we propose maximizing the ratio between the multiphoton signal and the n'th power of the excitation pulse energy as a criterion for optimal spectral filtering in SSFS when the soliton shows dramatic overlapping with the residual. This optimization is based on the most efficient signal generation and entirely depends on physical quantities that can be easily measured experimentally. Its application to MPM may reduce tissue damage, while maintaining high signal levels for efficient deep penetration.

  9. Intravital Multiphoton Imaging of the Kidney: Tubular Structure and Metabolism.

    PubMed

    Small, David M; Sanchez, Washington Y; Gobe, Glenda C

    2016-01-01

    Multiphoton microscopy (MPM) allows the visualization of dynamic pathophysiological events in real time in live animals. Intravital imaging can be applied to investigate novel mechanisms and treatments of different forms of kidney disease as well as improve our understanding of normal kidney physiology. Using rodent models, in conjunction with endogenous fluorescence and infused exogenous fluorescent dyes, measurement can be made of renal processes such as glomerular permeability, juxtaglomerular apparatus function, interactions of the tubulointerstitium, tubulovascular interactions, vascular flow rate, and the renin-angiotensin-aldosterone system. Subcellular processes including mitochondrial dynamics, reactive oxygen species production, cytosolic ion concentrations, and death processes of apoptosis and necrosis can also be seen and measured by MPM. The current methods chapter presents an overview of MPM with a focus on techniques for intravital kidney imaging and gives examples of instances where intravital MPM has been utilized to study renal pathophysiology. Suggestions are provided for MPM methods within the confines of intravital microscopy and selected kidney structure. MPM is undoubtedly a powerful new technique for application in experimental nephrology, and we believe it will continue to create new paradigms for understanding and treating kidney disease.

  10. High-throughput multiphoton-induced three-dimensional ablation and imaging for biotissues

    PubMed Central

    Lin, Chun-Yu; Li, Pei-Kao; Cheng, Li-Chung; Li, Yi-Cheng; Chang, Chia-Yuan; Chiang, Ann-Shyn; Dong, Chen Yuan; Chen, Shean-Jen

    2015-01-01

    In this study, a temporal focusing-based high-throughput multiphoton-induced ablation system with axially-resolved widefield multiphoton excitation has been successfully applied to rapidly disrupt biotissues. Experimental results demonstrate that this technique features high efficiency for achieving large-area laser ablation without causing serious photothermal damage in non-ablated regions. Furthermore, the rate of tissue processing can reach around 1.6 × 106 μm3/s in chicken tendon. Moreover, the temporal focusing-based multiphoton system can be efficiently utilized in optical imaging through iterating high-throughput multiphoton-induced ablation machining followed by widefield optical sectioning; hence, it has the potential to obtain molecular images for a whole bio-specimen. PMID:25780739

  11. Spectral imaging microscopy web sites and data.

    PubMed

    McNamara, George; Gupta, Amit; Reynaert, James; Coates, Thomas D; Boswell, Carl

    2006-08-01

    The Internet is enabling greater access to spectral imaging publications, spectral graphs, and data than that was available a generation ago. The spectral imaging systems discussed in this issue of Cytometry work because reagent and hardware spectra are reproducible, reusable, and provide input to spectral unmixing and spectral components recognition algorithms. These spectra need to be readily available in order to determine what to purchase, how to use it, and what the output means. We refer to several commercially sponsored and academic spectral web sites and discuss our spectral graphing and data sites. Sites include fluorescent dye graph servers from Invitrogen/Molecular Probes, BD Biosciences, Zeiss/Bio-Rad Cell Sciences, and filter set servers from Chroma Technology and Omega Optical. Several of these sites include data download capabilities. Recently, two microscope manufacturers have published on their web sites transmission curves for select objective lenses-crucial data for anyone doing multiphoton excitation microscopy. Notable among the academic sites, PhotoChemCAD 2.0 has over 200 dyes and a downloadable database/graphing program, and the USC-A Chemistry UV-vis Database displays absorption spectra of many dyes and indicators used in clinical histology and pathology. Our Fluorescent Spectra graphing/calculator site presents dyes, filters, and illumination data from many of these and additional sources. PubSpectra is our free download site which uses Microsoft Excel files as standardized human/machine readable format with over 2,000 biomedical spectra. The principle that data is not subject to copyright provides a framework in which all scientific data should be made freely accessible. PMID:16969821

  12. Spectral imaging microscopy web sites and data.

    PubMed

    McNamara, George; Gupta, Amit; Reynaert, James; Coates, Thomas D; Boswell, Carl

    2006-08-01

    The Internet is enabling greater access to spectral imaging publications, spectral graphs, and data than that was available a generation ago. The spectral imaging systems discussed in this issue of Cytometry work because reagent and hardware spectra are reproducible, reusable, and provide input to spectral unmixing and spectral components recognition algorithms. These spectra need to be readily available in order to determine what to purchase, how to use it, and what the output means. We refer to several commercially sponsored and academic spectral web sites and discuss our spectral graphing and data sites. Sites include fluorescent dye graph servers from Invitrogen/Molecular Probes, BD Biosciences, Zeiss/Bio-Rad Cell Sciences, and filter set servers from Chroma Technology and Omega Optical. Several of these sites include data download capabilities. Recently, two microscope manufacturers have published on their web sites transmission curves for select objective lenses-crucial data for anyone doing multiphoton excitation microscopy. Notable among the academic sites, PhotoChemCAD 2.0 has over 200 dyes and a downloadable database/graphing program, and the USC-A Chemistry UV-vis Database displays absorption spectra of many dyes and indicators used in clinical histology and pathology. Our Fluorescent Spectra graphing/calculator site presents dyes, filters, and illumination data from many of these and additional sources. PubSpectra is our free download site which uses Microsoft Excel files as standardized human/machine readable format with over 2,000 biomedical spectra. The principle that data is not subject to copyright provides a framework in which all scientific data should be made freely accessible.

  13. Multiphoton spectral analysis of benzo[a]pyrene uptake and metabolism in a rat liver cell line

    SciTech Connect

    Barhoumi, Rola; Mouneimne, Youssef; Ramos, Ernesto; Morisseau, Christophe; Hammock, Bruce D.; Safe, Stephen; Parrish, Alan R.; Burghardt, Robert C.

    2011-05-15

    Dynamic analysis of the uptake and metabolism of polycyclic aromatic hydrocarbons (PAHs) and their metabolites within live cells in real time has the potential to provide novel insights into genotoxic and non-genotoxic mechanisms of cellular injury caused by PAHs. The present work, combining the use of metabolite spectra generated from metabolite standards using multiphoton spectral analysis and an 'advanced unmixing process', identifies and quantifies the uptake, partitioning, and metabolite formation of one of the most important PAHs (benzo[a]pyrene, BaP) in viable cultured rat liver cells over a period of 24 h. The application of the advanced unmixing process resulted in the simultaneous identification of 8 metabolites in live cells at any single time. The accuracy of this unmixing process was verified using specific microsomal epoxide hydrolase inhibitors, glucuronidation and sulfation inhibitors as well as several mixtures of metabolite standards. Our findings prove that the two-photon microscopy imaging surpasses the conventional fluorescence imaging techniques and the unmixing process is a mathematical technique that seems applicable to the analysis of BaP metabolites in living cells especially for analysis of changes of the ultimate carcinogen benzo[a]pyrene-r-7,t-8-dihydrodiol-t-9,10-epoxide. Therefore, the combination of the two-photon acquisition with the unmixing process should provide important insights into the cellular and molecular mechanisms by which BaP and other PAHs alter cellular homeostasis.

  14. Real-time optical diagnosis of gastric cancer with serosal invasion using multiphoton imaging

    PubMed Central

    Yan, Jun; Zheng, Yu; Zheng, Xiaoling; Liu, Zhangyuanzhu; Liu, Wenju; Chen, Dexin; Dong, Xiaoyu; Li, Kai; Liu, Xiumin; Chen, Gang; Lu, Jianping; Chen, Jianxin; Zhuo, Shuangmu; Li, Guoxin

    2016-01-01

    A real-time optical biopsy, which could determine tissue histopathology, would be of extraordinary benefit to staging laparoscopy for gastric cancer with serosal invasion (T4) that requires downstage treatment. We investigated the feasibility of using multiphoton imaging to perform a real-time optical diagnosis of gastric cancer with or without serosal invasion. First, a pilot study was performed to establish the optical diagnostic features of gastric cancer with or without serosal invasion using multiphoton imaging compared with hematoxylin-eosin staining and Masson’s trichrome staining. Second, a blinded study was performed to compare the diagnostic sensitivity, specificity, and accuracy of multiphoton imaging and endoscopic ultrasonography (EUS) for T4 gastric cancer. In the pilot study, multiphoton imaging revealed collagen loss and degradation and cellular and nuclear pleomorphism in gastric cancer with serosal invasion. The collagen content in gastric cancer with or without serosal invasion was 0.36 ± 0.18 and 0.79 ± 0.16 (p < 0.001), respectively. In the blinded study, the sensitivity, specificity, and accuracy of EUS and multiphoton imaging for T4 gastric cancer were 70% and 90% (p = 0.029), 66.67% and 96.67% (p = 0.003), and 68.33% and 93.33% (p = 0.001), respectively. It is feasible to use multiphoton imaging to make a real-time optical diagnosis of gastric cancer with or without serosal invasion. PMID:27499365

  15. Multiphoton imaging microscopy at deeper layers with adaptive optics control of spherical aberration.

    PubMed

    Bueno, Juan M; Skorsetz, Martin; Palacios, Raquel; Gualda, Emilio J; Artal, Pablo

    2014-01-01

    Despite the inherent confocality and optical sectioning capabilities of multiphoton microscopy, three-dimensional (3-D) imaging of thick samples is limited by the specimen-induced aberrations. The combination of immersion objectives and sensorless adaptive optics (AO) techniques has been suggested to overcome this difficulty. However, a complex plane-by-plane correction of aberrations is required, and its performance depends on a set of image-based merit functions. We propose here an alternative approach to increase penetration depth in 3-D multiphoton microscopy imaging. It is based on the manipulation of the spherical aberration (SA) of the incident beam with an AO device while performing fast tomographic multiphoton imaging. When inducing SA, the image quality at best focus is reduced; however, better quality images are obtained from deeper planes within the sample. This is a compromise that enables registration of improved 3-D multiphoton images using nonimmersion objectives. Examples on ocular tissues and nonbiological samples providing different types of nonlinear signal are presented. The implementation of this technique in a future clinical instrument might provide a better visualization of corneal structures in living eyes.

  16. Label-free multiphoton imaging and photoablation of preinvasive cancer cells

    NASA Astrophysics Data System (ADS)

    Zhuo, Shuangmu; Chen, Jianxin; Wu, Guizhu; Zhu, Xiaoqin; Jiang, Xingshan; Xie, Shusen

    2012-01-01

    Detection and treatment of early lesions in epithelial tissue offer several possibilities for curing cancer, but it is challenging. Here, we present an optical technique, the combination of multiphoton imaging and absorption, to label-freely detect and ablate preinvasive cancer cells in epithelial tissue. We find that multiphoton imaging can label-freely visualize the principal features of nuclear atypia associated with epithelial precancerous lesions, and the spatial localization of multiphoton absorption can perform targeted ablation of preinvasive cancer cells with micrometer-sized volume precision. These results indicate that this optical technique has the capability to label-freely visualize and remove preinvasive cancer cells in epithelial tissue. This study highlights the potential of this technique as a "seek-and-treat" tool for early lesions in epithelial tissue.

  17. Multifocal multiphoton excitation and time correlated single photon counting detection for 3-D fluorescence lifetime imaging.

    PubMed

    Kumar, S; Dunsby, C; De Beule, P A A; Owen, D M; Anand, U; Lanigan, P M P; Benninger, R K P; Davis, D M; Neil, M A A; Anand, P; Benham, C; Naylor, A; French, P M W

    2007-10-01

    We report a multifocal multiphoton time-correlated single photon counting (TCSPC) fluorescence lifetime imaging (FLIM) microscope system that uses a 16 channel multi-anode PMT detector. Multiphoton excitation minimizes out-of-focus photobleaching, multifocal excitation reduces non-linear in-plane photobleaching effects and TCSPC electronics provide photon-efficient detection of the fluorescence decay profile. TCSPC detection is less prone to bleaching- and movement-induced artefacts compared to wide-field time-gated or frequency-domain FLIM. This microscope is therefore capable of acquiring 3-D FLIM images at significantly increased speeds compared to single beam multiphoton microscopy and we demonstrate this with live cells expressing a GFP tagged protein. We also apply this system to time-lapse FLIM of NAD(P)H autofluorescence in single live cells and report measurements on the change in the fluorescence decay profile following the application of a known metabolic inhibitor. PMID:19550524

  18. Three dimensional multiphoton imaging of fresh and whole mount developing mouse mammary glands

    PubMed Central

    2013-01-01

    Background The applications of multiphoton microscopy for deep tissue imaging in basic and clinical research are ever increasing, supplementing confocal imaging of the surface layers of cells in tissue. However, imaging living tissue is made difficult by the light scattering properties of the tissue, and this is extraordinarily apparent in the mouse mammary gland which contains a stroma filled with fat cells surrounding the ductal epithelium. Whole mount mammary glands stained with Carmine Alum are easily archived for later reference and readily viewed using bright field microscopy to observe branching architecture of the ductal network. Here, we report on the advantages of multiphoton imaging of whole mount mammary glands. Chief among them is that optical sectioning of the terminal end bud (TEB) and ductal epithelium allows the appreciation of abnormalities in structure that are very difficult to ascertain using either bright field imaging of the stained gland or the conventional approach of hematoxylin and eosin staining of fixed and paraffin-embedded sections. A second advantage is the detail afforded by second harmonic generation (SHG) in which collagen fiber orientation and abundance can be observed. Methods GFP-mouse mammary glands were imaged live or after whole mount preparation using a Zeiss LSM510/META/NLO multiphoton microscope with the purpose of obtaining high resolution images with 3D content, and evaluating any structural alterations induced by whole mount preparation. We describe a simple means for using a commercial confocal/ multiphoton microscope equipped with a Ti-Sapphire laser to simultaneously image Carmine Alum fluorescence and collagen fiber networks by SHG with laser excitation set to 860 nm. Identical terminal end buds (TEBs) were compared before and after fixation, staining, and whole mount preparation and structure of collagen networks and TEB morphologies were determined. Flexibility in excitation and emission filters was explored

  19. Differentiation of normal and cancerous lung tissues by multiphoton imaging

    NASA Astrophysics Data System (ADS)

    Wang, Chun-Chin; Li, Feng-Chieh; Wu, Ruei-Jhih; Hovhannisyan, Vladimir A.; Lin, Wei-Chou; Lin, Sung-Jan; So, Peter T. C.; Dong, Chen-Yuan

    2009-07-01

    We utilize multiphoton microscopy for the label-free diagnosis of noncancerous, lung adenocarcinoma (LAC), and lung squamous cell carcinoma (SCC) tissues from humans. Our results show that the combination of second-harmonic generation (SHG) and multiphoton excited autofluorescence (MAF) signals may be used to acquire morphological and quantitative information in discriminating cancerous from noncancerous lung tissues. Specifically, noncancerous lung tissues are largely fibrotic in structure, while cancerous specimens are composed primarily of tumor masses. Quantitative ratiometric analysis using MAF to SHG index (MAFSI) shows that the average MAFSI for noncancerous and LAC lung tissue pairs are 0.55+/-0.23 and 0.87+/-0.15, respectively. In comparison, the MAFSIs for the noncancerous and SCC tissue pairs are 0.50+/-0.12 and 0.72+/-0.13, respectively. Our study shows that nonlinear optical microscopy can assist in differentiating and diagnosing pulmonary cancer from noncancerous tissues.

  20. Clinical combination of multiphoton tomography and high frequency ultrasound imaging for evaluation of skin diseases

    NASA Astrophysics Data System (ADS)

    König, K.; Speicher, M.; Koehler, M. J.; Scharenberg, R.; Elsner, P.; Kaatz, M.

    2010-02-01

    For the first time, high frequency ultrasound imaging, multiphoton tomography, and dermoscopy were combined in a clinical study. Different dermatoses such as benign and malign skin cancers, connective tissue diseases, inflammatory skin diseases and autoimmune bullous skin diseases have been investigated with (i) state-of-the-art and highly sophisticated ultrasound systems for dermatology, (ii) the femtosecond-laser multiphoton tomograph DermaInspectTM and (iii) dermoscopes. Dermoscopy provides two-dimensional color imaging of the skin surface with a magnification up to 70x. Ultrasound images are generated from reflections of the emitted ultrasound signal, based on inhomogeneities of the tissue. These echoes are converted to electrical signals. Depending on the ultrasound frequency the penetration depth varies from about 1 mm to 16 mm in dermatological application. The 100-MHz-ultrasound system provided an axial resolution down to 16 μm and a lateral resolution down to 32 μm. In contrast to the wide-field ultrasound images, multiphoton tomography provided horizontal optical sections of 0.36×0.36 mm2 down to 200 μm tissue depth with submicron resolution. The autofluorescence of mitochondrial coenzymes, melanin, and elastin as well as the secondharmonic- generation signal of the collagen network were imaged. The combination of ultrasound and multiphoton tomography provides a novel opportunity for diagnostics of skin disorders.

  1. Stepwise multi-photon activation fluorescence reveals a new method of melanoma imaging for dermatologists

    NASA Astrophysics Data System (ADS)

    Lai, Zhenhua; Lian, Christine; Ma, Jie; Yu, Jingyi; Gu, Zetong; Rajadhyaksha, Milind; DiMarzio, Charles A.

    2014-02-01

    Previous research has shown that the stepwise multi-photon activated fluorescence (SMPAF) of melanin, activated by a continuous-wave (CW) mode near infrared (NIR) laser, is a low cost and reliable method of detecting melanin. SMPAF images of melanin in a mouse hair and a formalin fixed mouse melanoma were compared with conventional multiphoton fluorescence microscopy (MPFM) images and confocal reflectance microscopy (CRM) images, all of which were acquired at an excitation wavelength of 920 nm, to further prove the effectiveness of SMPAF in detecting melanin. SMPAF images add specificity for melanin detection to MPFM images and CRM images. Melanin SMPAF can be a promising technology to enable melanoma imaging for dermatologists.

  2. Differentiation of normal and cancerous lung tissues by multiphoton imaging

    NASA Astrophysics Data System (ADS)

    Wang, Chun-Chin; Li, Feng-Chieh; Wu, Ruei-Jr; Hovhannisyan, Vladimir A.; Lin, Wei-Chou; Lin, Sung-Jan; So, Peter T. C.; Dong, Chen-Yuan

    2010-02-01

    In this work, we utilized multiphoton microscopy for the label-free diagnosis of non-cancerous, lung adenocarcinoma (LAC), and lung squamous cell carcinoma (SCC) tissues from human. Our results show that the combination of second harmonic generation (SHG) and multiphoton excited autofluorescence (MAF) signals may be used to acquire morphological and quantitative information in discriminating cancerous from non-cancerous lung tissues. Specifically, non-cancerous lung tissues are largely fibrotic in structure while cancerous specimens are composed primarily of tumor masses. Quantitative ratiometric analysis using MAF to SHG index (MAFSI or SAAID) shows that the average MAFSI for noncancerous and LAC lung tissue pairs are 0.55 +/-0.23 and 0.87+/-0.15 respectively. In comparison, the MAFSIs for the noncancerous and SCC tissue pairs are 0.50+/-0.12 and 0.72+/-0.13 respectively. Intrinsic fluorescence ratio (FAD/NADH) of SCC and non-cancerous tissues are 0.40+/-0.05 and 0.53+/-0.05 respectively, the redox ratio of SCC diminishes significantly, indicating that increased cellular metabolic activity. Our study shows that nonlinear optical microscopy can assist in differentiating and diagnosing pulmonary cancer from non-cancerous tissues. With additional development, multiphoton microscopy may be used for the clinical diagnosis of lung cancers.

  3. Clinical multiphoton FLIM tomography

    NASA Astrophysics Data System (ADS)

    König, Karsten

    2012-03-01

    This paper gives an overview on current clinical high resolution multiphoton fluorescence lifetime imaging in volunteers and patients. Fluorescence lifetime imaging (FLIM) in Life Sciences was introduced in Jena/Germany in 1988/89 based on a ZEISS confocal picosecond dye laser scanning microscope equipped with a single photon counting unit. The porphyrin distribution in living cells and living tumor-bearing mice was studied with high spatial, temporal, and spectral resolution. Ten years later, time-gated cameras were employed to detect dental caries in volunteers based on one-photon excitation of autofluorescent bacteria with long fluorescence lifetimes. Nowadays, one-photon FLIM based on picosecond VIS laser diodes are used to study ocular diseases in humans. Already one decade ago, first clinical twophoton FLIM images in humans were taken with the certified clinical multiphoton femtosecond laser tomograph DermaInspectTM. Multiphoton tomographs with FLIM modules are now operating in hospitals at Brisbane, Tokyo, Berlin, Paris, London, Modena and other European cities. Multiple FLIM detectors allow spectral FLIM with a temporal resolution down to 20 ps (MCP) / 250 ps (PMT) and a spectral resolution of 10 nm. Major FLIM applications include the detection of intradermal sunscreen and tattoo nanoparticles, the detection of different melanin types, the early diagnosis of dermatitis and malignant melanoma, as well as the measurement of therapeutic effects in pateints suffering from dermatitis. So far, more than 1,000 patients and volunteers have been investigated with the clinical multiphoton FLIM tomographs DermaInspectTM and MPTflexTM.

  4. A high speed multifocal multiphoton fluorescence lifetime imaging microscope for live-cell FRET imaging

    PubMed Central

    Poland, Simon P.; Krstajić, Nikola; Monypenny, James; Coelho, Simao; Tyndall, David; Walker, Richard J.; Devauges, Viviane; Richardson, Justin; Dutton, Neale; Barber, Paul; Li, David Day-Uei; Suhling, Klaus; Ng, Tony; Henderson, Robert K.; Ameer-Beg, Simon M.

    2015-01-01

    We demonstrate diffraction limited multiphoton imaging in a massively parallel, fully addressable time-resolved multi-beam multiphoton microscope capable of producing fluorescence lifetime images with sub-50ps temporal resolution. This imaging platform offers a significant improvement in acquisition speed over single-beam laser scanning FLIM by a factor of 64 without compromising in either the temporal or spatial resolutions of the system. We demonstrate FLIM acquisition at 500 ms with live cells expressing green fluorescent protein. The applicability of the technique to imaging protein-protein interactions in live cells is exemplified by observation of time-dependent FRET between the epidermal growth factor receptor (EGFR) and the adapter protein Grb2 following stimulation with the receptor ligand. Furthermore, ligand-dependent association of HER2-HER3 receptor tyrosine kinases was observed on a similar timescale and involved the internalisation and accumulation or receptor heterodimers within endosomes. These data demonstrate the broad applicability of this novel FLIM technique to the spatio-temporal dynamics of protein-protein interaction. PMID:25780724

  5. Label-free imaging and quantitative chemical analysis of Alzheimer's disease brain samples with multimodal multiphoton nonlinear optical microspectroscopy

    NASA Astrophysics Data System (ADS)

    Lee, Jang Hyuk; Kim, Dae Hwan; Song, Woo Keun; Oh, Myoung-Kyu; Ko, Do-Kyeong

    2015-05-01

    We developed multimodal multiphoton microspectroscopy using a small-diameter probe with gradient-index lenses and applied it to unstained Alzheimer's disease (AD) brain samples. Our system maintained the image quality and spatial resolution of images obtained using an objective lens of similar numerical aperture. Multicolor images of AD brain samples were obtained simultaneously by integrating two-photon excited fluorescence and second-harmonic generation on a coherent anti-Stokes Raman scattering (CARS) microendoscope platform. Measurements of two hippocampal regions, the cornus ammonis-1 and dentate gyrus, revealed more lipids, amyloid fibers, and collagen in the AD samples than in the normal samples. Normal and AD brains were clearly distinguished by a large spectral difference and quantitative analysis of the CH mode using CARS microendoscope spectroscopy. We expect this system to be an important diagnosis tool in AD research.

  6. Label-free imaging and quantitative chemical analysis of Alzheimer's disease brain samples with multimodal multiphoton nonlinear optical microspectroscopy.

    PubMed

    Lee, Jang Hyuk; Kim, Dae Hwan; Song, Woo Keun; Oh, Myoung-Kyu; Ko, Do-Kyeong

    2015-05-01

    We developed multimodal multiphoton microspectroscopy using a small-diameter probe with gradient-index lenses and applied it to unstained Alzheimer's disease (AD) brain samples. Our system maintained the image quality and spatial resolution of images obtained using an objective lens of similar numerical aperture. Multicolor images of AD brain samples were obtained simultaneously by integrating two-photon excited fluorescence and second-harmonic generation on a coherent anti-Stokes Raman scattering (CARS) microendoscope platform. Measurements of two hippocampal regions, the cornus ammonis-1 and dentate gyrus, revealed more lipids, amyloid fibers, and collagen in the AD samples than in the normal samples. Normal and AD brains were clearly distinguished by a large spectral difference and quantitative analysis of the CH mode using CARS microendoscope spectroscopy. We expect this system to be an important diagnosis tool in AD research

  7. Focal switching of photochromic fluorescent proteins enables multiphoton microscopy with superior image contrast

    PubMed Central

    Kao, Ya-Ting; Zhu, Xinxin; Xu, Fang; Min, Wei

    2012-01-01

    Probing biological structures and functions deep inside live organisms with light is highly desirable. Among the current optical imaging modalities, multiphoton fluorescence microscopy exhibits the best contrast for imaging scattering samples by employing a spatially confined nonlinear excitation. However, as the incident laser power drops exponentially with imaging depth into the sample due to the scattering loss, the out-of-focus background eventually overwhelms the in-focus signal, which defines a fundamental imaging-depth limit. Herein we significantly improve the image contrast for deep scattering samples by harnessing reversibly switchable fluorescent proteins (RSFPs) which can be cycled between bright and dark states upon light illumination. Two distinct techniques, multiphoton deactivation and imaging (MPDI) and multiphoton activation and imaging (MPAI), are demonstrated on tissue phantoms labeled with Dronpa protein. Such a focal switch approach can generate pseudo background-free images. Conceptually different from wave-based approaches that try to reduce light scattering in turbid samples, our work represents a molecule-based strategy that focused on imaging probes. PMID:22876358

  8. Coupling CARS with multiphoton fluorescence and high harmonic generation imaging modalities using a femtosecond laser source

    NASA Astrophysics Data System (ADS)

    Chen, Hongtao; Slipchenko, Mikhail N.; Zhu, Jiabin; Buhman, Kimberly K.; Cheng, Ji-Xin

    2009-02-01

    Multimodal nonlinear optical imaging has opened new opportunities and becomes a powerful tool for imaging complex tissue samples with inherent 3D spatial resolution.. We present a robust and easy-to-operate approach to add the coherent anti-stokes Raman scattering (CARS) imaging modality to a widely used multiphoton microscope. The laser source composed of a Mai Tai femtosecond laser and an optical parametric oscillator (OPO) offers one-beam, two-beam and three-beam modalities. The Mai Tai output at 790 nm is split into two beams, with 80% of the power being used to pump the OPO. The idler output at 2036 nm from OPO is doubled using a periodically poled lithium niobate (PPLN) crystal. This frequency-doubled idler beam at 1018 nm is sent through a delay line and collinearly combined with the other Mai Tai beam for CARS imaging on a laser-scanning microscope. This Mai Tai beam is also used for multiphoton fluorescence and second harmonic generation (SHG) imaging. The signal output at 1290 nm from OPO is used for SHG and third-harmonic generation (THG) imaging. External detectors are installed for both forward and backward detection, whereas two internal lamda-scan detectors are employed for microspectroscopy analysis. This new system allows vibrationally resonant CARS imaging of lipid bodies, SHG imaging of collagen fibers, and multiphoton fluorescence analysis in fresh tissues. As a preliminary application, the effect of diacylglycerol acyltransferase 1 (DGAT1) deficiency on liver lipid metabolism in mice was investigated.

  9. Nanoparticle-assisted-multiphoton microscopy for in vivo brain imaging of mice

    NASA Astrophysics Data System (ADS)

    Qian, Jun

    2015-03-01

    Neuro/brain study has attracted much attention during past few years, and many optical methods have been utilized in order to obtain accurate and complete neural information inside the brain. Relying on simultaneous absorption of two or more near-infrared photons by a fluorophore, multiphoton microscopy can achieve deep tissue penetration and efficient light detection noninvasively, which makes it very suitable for thick-tissue and in vivo bioimaging. Nanoparticles possess many unique optical and chemical properties, such as anti-photobleaching, large multiphoton absorption cross-section, and high stability in biological environment, which facilitates their applications in long-term multiphoton microscopy as contrast agents. In this paper, we will introduce several typical nanoparticles (e.g. organic dye doped polymer nanoparticles and gold nanorods) with high multiphoton fluorescence efficiency. We further applied them in two- and three-photon in vivo functional brain imaging of mice, such as brain-microglia imaging, 3D architecture reconstruction of brain blood vessel, and blood velocity measurement.

  10. Enabling Multiphoton and Second Harmonic Generation Imaging in Paraffin-Embedded and Histologically Stained Sections

    PubMed Central

    Monaghan, Michael G.; Kroll, Sebastian; Brucker, Sara Y.

    2016-01-01

    Nonlinear microscopy, namely multiphoton imaging and second harmonic generation (SHG), is an established noninvasive technique useful for the imaging of extracellular matrix (ECM). Typically, measurements are performed in vivo on freshly excised tissues or biopsies. In this article, we describe the effect of rehydrating paraffin-embedded sections on multiphoton and SHG emission signals and the acquisition of nonlinear images from hematoxylin and eosin (H&E)-stained sections before and after a destaining protocol. Our results reveal that bringing tissue sections to a physiological state yields a significant improvement in nonlinear signals, particularly in SHG. Additionally, the destaining of sections previously processed with H&E staining significantly improves their SHG emission signals during imaging, thereby allowing sufficient analysis of collagen in these sections. These results are important for researchers and pathologists to obtain additional information from paraffin-embedded tissues and archived samples to perform retrospective analysis of the ECM or gain additional information from rare samples. PMID:27018844

  11. Multiphoton imaging of upconverting lanthanide nanoparticles in three dimensional models of cancer

    NASA Astrophysics Data System (ADS)

    Gainer, Christian F.; Romanowski, Marek

    2013-02-01

    While upconverting lanthanide nanoparticles have numerous advantages over other exogenous contrast agents used in scanned multiphoton imaging, their long luminescence lifetimes cause images collected with non-descanned detection to be greatly blurred. We demonstrate herein the use of Richardson-Lucy deconvolution to deblur luminescence images obtained via multiphoton scanning microscopy. Images were taken of three dimensional models of colon and ovarian cancer following incubation with NaYF4:Yb,Er nanoparticles functionalized with an antibody for EGFR and folic acid respectively. Following deconvolution, images had a lateral resolution on par with the optimal performance of the imaging system used, ~1.2 μm, and an axial resolution of ~5 μm. Due to the relatively high multiphoton excitation efficiency of these nanoparticles, it is possible to follow binding of individual particles in tissue. In addition, their extreme photostability allows for prolonged imaging without significant loss in luminescence signal. With these advantageous properties in mind, we also discuss the potential application of upconverting lanthanide nanoparticles for tracking of specific, cancer relevant receptors in tissue.

  12. Image segmentation for integrated multiphoton microscopy and reflectance confocal microscopy imaging of human skin in vivo

    PubMed Central

    Chen, Guannan; Lui, Harvey

    2015-01-01

    Background Non-invasive cellular imaging of the skin in vivo can be achieved in reflectance confocal microscopy (RCM) and multiphoton microscopy (MPM) modalities to yield complementary images of the skin based on different optical properties. One of the challenges of in vivo microscopy is the delineation (i.e., segmentation) of cellular and subcellular architectural features. Methods In this work we present a method for combining watershed and level-set models for segmentation of multimodality images obtained by an integrated MPM and RCM imaging system from human skin in vivo. Results Firstly, a segmentation model based on watershed is introduced for obtaining the accurate structure of cell borders from the RCM image. Secondly,, a global region based energy level-set model is constructed for extracting the nucleus of each cell from the MPM image. Thirdly, a local region-based Lagrange Continuous level-set approach is used for segmenting cytoplasm from the MPM image. Conclusions Experimental results demonstrated that cell borders from RCM image and boundaries of cytoplasm and nucleus from MPM image can be obtained by our segmentation method with better accuracy and effectiveness. We are planning to use this method to perform quantitative analysis of MPM and RCM images of in vivo human skin to study the variations of cellular parameters such as cell size, nucleus size and other mophormetric features with skin pathologies. PMID:25694949

  13. In vivo imaging of unstained tissues using a compact and flexible multiphoton microendoscope

    NASA Astrophysics Data System (ADS)

    Brown, Christopher M.; Rivera, David R.; Pavlova, Ina; Ouzounov, Dimitre G.; Williams, Wendy O.; Mohanan, Sunish; Webb, Watt W.; Xu, Chris

    2012-04-01

    We use a compact and flexible multiphoton microendoscope (MPME) to acquire in vivo images of unstained liver, kidney, and colon from an anesthetized rat. The device delivers femtosecond pulsed 800 nm light from the core of a raster-scanned dual-clad fiber (DCF), which is focused by a miniaturized gradient-index lens assembly into tissue. Intrinsic fluorescence and second-harmonic generation signal from the tissue is epi-collected through the core and inner clad of the same DCF. The MPME has a rigid distal tip of 3 mm in outer diameter and 4 cm in length. The image field-of-view measures 115 μm by 115 μm and was acquired at 4.1 frames/s with 75 mW illumination power at the sample. Organs were imaged after anesthetizing Sprague-Dawley rats with isofluorane gas, accessing tissues via a ventral-midline abdominal incision, and isolating the organs with tongue depressors. In vivo multiphoton images acquired from liver, kidney, and colon using this device show features similar to that of conventional histology slides, without motion artifact, in ~75% of imaged frames. To the best of our knowledge, this is the first demonstration of multiphoton imaging of unstained tissue from a live subject using a compact and flexible MPME device.

  14. In vivo stepwise multi-photon activation fluorescence imaging of melanin in human skin

    NASA Astrophysics Data System (ADS)

    Lai, Zhenhua; Gu, Zetong; Abbas, Saleh; Lowe, Jared; Sierra, Heidy; Rajadhyaksha, Milind; DiMarzio, Charles

    2014-03-01

    The stepwise multi-photon activated fluorescence (SMPAF) of melanin is a low cost and reliable method of detecting melanin because the activation and excitation can be a continuous-wave (CW) mode near infrared (NIR) laser. Our previous work has demonstrated the melanin SMPAF images in sepia melanin, mouse hair, and mouse skin. In this study, we show the feasibility of using SMPAF to detect melanin in vivo. in vivo melanin SMPAF images of normal skin and benign nevus are demonstrated. SMPAF images add specificity for melanin detection than MPFM images and CRM images. Melanin SMPAF is a promising technology to enable early detection of melanoma for dermatologists.

  15. Spectral imaging using forward-viewing spectrally encoded endoscopy.

    PubMed

    Zeidan, Adel; Yelin, Dvir

    2016-02-01

    Spectrally encoded endoscopy (SEE) enables miniature, small-diameter endoscopic probes for minimally invasive imaging; however, using the broadband spectrum to encode space makes color and spectral imaging nontrivial and challenging. By careful registration and analysis of image data acquired by a prototype of a forward-viewing dual channel spectrally encoded rigid probe, we demonstrate spectral and color imaging within a narrow cylindrical lumen. Spectral imaging of calibration cylindrical test targets and an ex-vivo blood vessel demonstrates high-resolution spatial-spectral imaging with short (10 μs/line) exposure times. PMID:26977348

  16. Spectral imaging using forward-viewing spectrally encoded endoscopy

    PubMed Central

    Zeidan, Adel; Yelin, Dvir

    2016-01-01

    Spectrally encoded endoscopy (SEE) enables miniature, small-diameter endoscopic probes for minimally invasive imaging; however, using the broadband spectrum to encode space makes color and spectral imaging nontrivial and challenging. By careful registration and analysis of image data acquired by a prototype of a forward-viewing dual channel spectrally encoded rigid probe, we demonstrate spectral and color imaging within a narrow cylindrical lumen. Spectral imaging of calibration cylindrical test targets and an ex-vivo blood vessel demonstrates high-resolution spatial-spectral imaging with short (10 μs/line) exposure times. PMID:26977348

  17. Rayleigh imaging in spectral mammography

    NASA Astrophysics Data System (ADS)

    Berggren, Karl; Danielsson, Mats; Fredenberg, Erik

    2016-03-01

    Spectral imaging is the acquisition of multiple images of an object at different energy spectra. In mammography, dual-energy imaging (spectral imaging with two energy levels) has been investigated for several applications, in particular material decomposition, which allows for quantitative analysis of breast composition and quantitative contrast-enhanced imaging. Material decomposition with dual-energy imaging is based on the assumption that there are two dominant photon interaction effects that determine linear attenuation: the photoelectric effect and Compton scattering. This assumption limits the number of basis materials, i.e. the number of materials that are possible to differentiate between, to two. However, Rayleigh scattering may account for more than 10% of the linear attenuation in the mammography energy range. In this work, we show that a modified version of a scanning multi-slit spectral photon-counting mammography system is able to acquire three images at different spectra and can be used for triple-energy imaging. We further show that triple-energy imaging in combination with the efficient scatter rejection of the system enables measurement of Rayleigh scattering, which adds an additional energy dependency to the linear attenuation and enables material decomposition with three basis materials. Three available basis materials have the potential to improve virtually all applications of spectral imaging.

  18. Using multiphoton fluorescence lifetime imaging to characterize liver damage and fluorescein disposition in liver in vivo

    NASA Astrophysics Data System (ADS)

    Thorling, Camilla A.; Studier, Hauke; Crawford, Darrell; Roberts, Michael S.

    2016-03-01

    Liver disease is the fifth most common cause of death and unlike many other major causes of mortality, liver disease rates are increasing rather than decreasing. There is no ideal measurement of liver disease and although biopsies are the gold standard, this only allows for a spot examination and cannot follow dynamic processes of the liver. Intravital imaging has the potential to extract detailed information over a larger sampling area continuously. The aim of this project was to investigate whether multiphoton and fluorescence lifetime imaging microscopy could detect early liver damage and to assess whether it could detect changes in metabolism of fluorescein in normal and diseased livers. Four experimental groups were used in this study: 1) control; 2) ischemia reperfusion injury; 3) steatosis and 4) steatosis with ischemia reperfusion injury. Results showed that multiphoton microscopy could visualize morphological changes such as decreased fluorescence of endogenous fluorophores and the presence of lipid droplets, characteristic of steatosis. Fluorescence lifetime imaging microscopy showed increase in NADPH in steatosis with and without ischemia reperfusion injury and could detect changes in metabolism of fluorescein to fluorescein monoglurcuronide, which was impaired in steatosis with ischemia reperfusion injury. These results concluded that the combination of multiphoton microscopy and fluorescence lifetime imaging is a promising method of assessing early stage liver damage and that it can be used to study changes in drug metabolism in the liver as an indication of liver disease and has the potential to replace the traditional static liver biopsy currently used.

  19. Spectral ensemble ghost imaging

    NASA Astrophysics Data System (ADS)

    Jha, Nandan

    2015-07-01

    In the last few years, the field of ghost imaging has seen many new developments. From computational ghost imaging to 3D gh.ost imaging, this field has shown many interesting applications. But the method of obtaining an image in ghost imaging experiments still requires data to be recorded over a long duration of time due to averaging over many shots of data. We propose a method to get the intensity correlated images in one shot by averaging over different wavelength components rather than different time components. We derive the maximum number of wavelength components that can be used for a given multi-wavelength source. The applicability of this scheme is finally verified by numerical simulations.

  20. NI-78LABEL-FREE MULTIPHOTON MICROSCOPY: A NOVEL TOOL FOR THE IMAGING OF BRAIN TUMORS

    PubMed Central

    Uckermann, Ortrud; Galli, Roberta; Geiger, Kathrin; Koch, Edmund; Schackert, Gabriele; Steiner, Gerald; Kirsch, Matthias

    2014-01-01

    Changes in tissue composition caused by brain tumor growth involve a series of complex biochemical alterations which can be imaged on unstained native tissue using multiphoton microscopy: We used coherent anti-Stokes Raman scattering (CARS) imaging that resonantly excites the symmetric stretching vibration of CH2 groups at 2850 cm−1 and visualizes lipid content in combination with imaging of endogenous two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) to discern different types of tumors from normal tissue in unstained, native brain samples. Experimental brain tumors were induced in nude mice NMRI nu/nu (n = 25) by stereotactic implantation of glioblastoma (U87), melanoma (A375) and breast cancer (MCF-7) cell lines. Label-free multiphoton microscopy of brain cryosections provided exhaustive information of the tumor morphochemistry. The tumor border was defined with cellular resolution by a strong reduction of CARS signal intensity to 61% (glioblastoma), 71% (melanoma) and 68% (breast cancer). This reduction of lipid content within the tumor was confirmed by Raman spectroscopy. Micrometastases infiltrating normal tissue (size 50 - 200 µm) were identified in glioblastoma and melanoma. Additionally, multiphoton microscopy proved a reduction of CARS signal intensity in all human glioblastoma samples analyzed (to 72%, n = 6). Additionally, relevant SHG and TPEF signals were detected in human primary and secondary brain tumor samples and enabled to image variations in tumor associated vasculature, fibrosis, necrosis and nuclear size and density. All primary or secondary brain tumors investigated were characterized by a lower intensity of the CARS signal, therefore offering a simple tool for objective tumor detection and delineation. The combination of techniques allows retrieving a quantity of information on native unstained tissue which is comparable to H&E staining. Therefore, label-free multiphoton microscopy has the potential to become a

  1. Compact fixed wavelength femtosecond oscillators for multi-photon imaging

    NASA Astrophysics Data System (ADS)

    Hakulinen, T.; Klein, J.; Zadoyan, R.; Baldacchini, T.; Franke, T.

    2015-03-01

    In recent years two-photon microscopy with fixed-wavelength has raised increasing interest in life-sciences: Two-photon (2P) absorption spectra of common dyes are broader than single-photon ones. Therefore, excitation of several dyes simultaneously with a single IR laser wavelength is feasible and could be seen as an advantage in 2P microscopy. We used pulsed fixed-wavelength infrared lasers with center wavelength at 1040 nm, for two-photon microscopy in a variety of biologically relevant samples, among these a mouse brain sample, a mouse artery (within the animal, acute preparation), and a preparation of mouse bladder. The 1040 nm laser proved to be efficient not only in exciting fluorescence from yellow fluorescent protein (YFP) and red fluorescent dyes, but also for second harmonic generation (SHG) signals from muscle tissue and collagen. With this work we demonstrate that economical, small-footprint fixedwavelength lasers can present an interesting alternative to tunable lasers that are commonly used in multiphoton microscopy.

  2. Multiphoton fluorescence imaging of NADH to quantify metabolic changes in epileptic tissue in vitro

    NASA Astrophysics Data System (ADS)

    Chia, Thomas H.; Zinter, Joseph; Spencer, Dennis D.; Williamson, Anne; Levene, Michael J.

    2007-02-01

    A powerful advantage of multiphoton microscopy is its ability to image endogenous fluorophores such as the ubiquitous coenzyme NADH in discrete cellular populations. NADH is integral in both oxidative and non-oxidative cellular metabolism. NADH loses fluorescence upon oxidation to NAD +; thus changes in NADH fluorescence can be used to monitor metabolism. Recent studies have suggested that hypo metabolic astrocytes play an important role in cases of temporal lobe epilepsy (TLE). Current theories suggest this may be due to defective and/or a reduced number of mitochondria or dysfunction of the neuronal-astrocytic metabolic coupling. Measuring NADH fluorescence changes following chemical stimulation enables the quantification of the cellular distribution of metabolic anomalies in epileptic brain tissue compared to healthy tissue. We present what we believe to be the first multiphoton microscopy images of NADH from the human brain. We also present images of NADH fluorescence from the hippocampus of the kainate-treated rat TLE model. In some experiments, human and rat astrocytes were selectively labeled with the fluorescent dye sulforhodamine 101 (SR101). Our results demonstrate that multiphoton microscopy is a powerful tool for assaying the metabolic pathologies associated with temporal lobe epilepsy in humans and in rodent models.

  3. In vivo multiphoton imaging of collagen remodeling after microablative fractional rejuvenation

    NASA Astrophysics Data System (ADS)

    Cicchi, Riccardo; Kapsokalyvas, Dimitrios; Troiano, Michela; Campolmi, Piero; Morini, Cristiano; Cosci, Alessandro; Massi, Daniela; Lotti, Torello; Pavone, Francesco S.

    2011-03-01

    The potential of multiphoton microscopy in providing in-vivo early diagnosis of skin lesions has already been demonstrated, while its capability in therapy follow-up has not been deeply explored so far. Two-photon excited fluorescence and second-harmonic generation microscopy were used in combination to follow-up collagen remodeling after laser micro-ablative rejuvenation. Treated regions of volunteers were imaged with multiphoton microscopy before and after treatment, and we found a strong age-dependence of the treatment effectiveness. In particular, the photorejuvenating effect was negligible in young subjects (< 30 years), whereas a significant production of new collagen was observed in aged subjects (> 70 years). Quantification of the amount of newly produced collagen and its organization were performed by means of visual examination of two-photon images. The obtained results demonstrate the performance of laser fractional micro-ablative rejuvenation without the need of an invasive biopsy as well as the wide applicability range of applications for multiphoton microscopy in clinical dermatology.

  4. Eliminating the scattering ambiguity in multifocal, multimodal multiphoton imaging systems

    PubMed Central

    Hoover, Erich E.; Field, Jeffrey J.; Winters, David G.; Young, Michael D.; Chandler, Eric V.; Speirs, John C.; Kim, Susy M.; Ding, Shi-you; Bartels, Randy A.; Wang, Jing W.; Squier, Jeff A.

    2013-01-01

    Four images of Drosophila Melanogaster antennal lobe structure labeled with red fluorescent protein. The images are separated axially by 7 μm in depth, and were all acquired simultaneously from a single-element detector. PMID:22461190

  5. Imaging NO elimination in the infrared multiphoton dissociation of nitroalkanes and alkyl nitrites

    NASA Astrophysics Data System (ADS)

    Fernando, Ravin; Ariyasingha, Nuwandi M.; Suits, Arthur G.

    2016-02-01

    We present a DC slice imaging study of the decomposition of C2, C3 and C4 nitroalkanes and alkyl nitrites, focusing on the NO elimination channel, possibly a minor pathway. Infrared multiphoton dissociation (IRMPD) is used to induce dissociation on the ground electronic state under collisionless conditions. The channels that produced NO as a product were studied and compared among the target molecules to gain a better understanding of the isomerization of the nitroalkanes prior to dissociation. Trends in the total translational energy and NO rotational temperatures obtained from the images are discussed.

  6. In vivo multiphoton imaging of the cornea: polarization-resolved second harmonic generation from stromal collagen

    NASA Astrophysics Data System (ADS)

    Latour, G.; Gusachenko, I.; Kowalczuk, L.; Lamarre, I.; Schanne-Klein, M.-C.

    2012-03-01

    Multiphoton microscopy provides specific and contrasted images of unstained collagenous tissues such as tendons or corneas. Polarization-resolved second harmonic generation (SHG) measurements have been implemented in a laserscanning multiphoton microscope. Distortion of the polarimetric response due to birefringence and diattenuation during propagation of the laser excitation has been shown in rat-tail tendons. A model has been developed to account for these effects and correct polarization-resolved SHG images in thick tissues. This new modality is then used in unstained human corneas to access two quantitative parameters: the fibrils orientation within the collagen lamellae and the ratio of the main second-order nonlinear tensorial components. Orientation maps obtained from polarization resolution of the trans-detected SHG images are in good agreement with the striated features observed in the raw images. Most importantly, polarization analysis of the epi-detected SHG images also enables to map the fibrils orientation within the collagen lamellae while epi-detected SHG images of corneal stroma are spatially homogenous and do not enable direct visualization of the fibrils orientation. Depth profiles of the polarimetric SHG response are also measured and compared to models accounting for orientation changes of the collagen lamellae within the focal volume. Finally, in vivo polarization-resolved SHG is performed in rat corneas and structural organization of corneal stroma is determined using epi-detected signals.

  7. Multiphoton imaging of quantum dot bioconjugates in cultured cells following Nd:YLF laser excitation

    NASA Astrophysics Data System (ADS)

    Serrano, Elba E.; Knight, V. B.

    2005-04-01

    Quantum dot bioconjugates offer unprecedented opportunities for monitoring biological processes and molecular interactions in cells, tissues, and organs. We are interested in developing applications that permit investigation of physiological processes and cytoskeletal organization in live cells, and allow imaging of complex organs, such as the auditory and vestibular sensory structures of the inner ear. Multiphoton microscopy is a powerful technique for acquiring images from deep within a sample while reducing phototoxic effects of laser light exposure on cells. Previous studies have established that a solid-state Nd:YLF laser can be used to acquire two-photon and three-photon images from live cells while minimizing phototoxic side effects (Wokosin et al., 1996, Bioimaging, 4:208-214; Squirrell et al., 1999, Nature Biotechnology, 8:763-767). We present here the results of experiments using an all-solid-state Nd:YLF 1047 nm femtosecond laser (Microlase DPM1000) source to excite quantum dot bioconjugates. Cells were labeled with Qdot (Quantum Dot Corporation) bioconjugates or with Alexa Fluor (Molecular Probes) bioconjugates and then imaged with a BioRad 1024 confocal microscope configured for multiphoton imaging using internal or external (non-descanned) detectors. Results demonstrate that the Nd:YLF laser can be used to stimulate fluorescence emission of quantum dots and Alexa Fluor bioconjugates in cultured amphibian (Xenopus) and mammalian (rat, chinese hamster) cells. We conclude that the Nd:YLF laser is a viable excitation source that extends the applicability of quantum dots for investigation of biological processes using multiphoton microscopy.

  8. Optimizing ultrafast illumination for multiphoton-excited fluorescence imaging.

    PubMed

    Stoltzfus, Caleb R; Rebane, Aleksander

    2016-05-01

    We study the optimal conditions for high throughput two-photon excited fluorescence (2PEF) and three-photon excited fluorescence (3PEF) imaging using femtosecond lasers. We derive relations that allow maximization of the rate of imaging depending on the average power, pulse repetition rate, and noise characteristics of the laser, as well as on the size and structure of the sample. We perform our analysis using ~100 MHz, ~1 MHz and 1 kHz pulse rates and using both a tightly-focused illumination beam with diffraction-limited image resolution, as well loosely focused illumination with a relatively low image resolution, where the latter utilizes separate illumination and fluorescence detection beam paths. Our theoretical estimates agree with the experiments, which makes our approach especially useful for optimizing high throughput imaging of large samples with a field-of-view up to 10x10 cm(2). PMID:27231620

  9. Optimizing ultrafast illumination for multiphoton-excited fluorescence imaging

    PubMed Central

    Stoltzfus, Caleb R.; Rebane, Aleksander

    2016-01-01

    We study the optimal conditions for high throughput two-photon excited fluorescence (2PEF) and three-photon excited fluorescence (3PEF) imaging using femtosecond lasers. We derive relations that allow maximization of the rate of imaging depending on the average power, pulse repetition rate, and noise characteristics of the laser, as well as on the size and structure of the sample. We perform our analysis using ~100 MHz, ~1 MHz and 1 kHz pulse rates and using both a tightly-focused illumination beam with diffraction-limited image resolution, as well loosely focused illumination with a relatively low image resolution, where the latter utilizes separate illumination and fluorescence detection beam paths. Our theoretical estimates agree with the experiments, which makes our approach especially useful for optimizing high throughput imaging of large samples with a field-of-view up to 10x10 cm2. PMID:27231620

  10. Fast volumetric imaging with patterned illumination via digital micro-mirror device-based temporal focusing multiphoton microscopy

    PubMed Central

    Chang, Chia-Yuan; Hu, Yvonne Yuling; Lin, Chun-Yu; Lin, Cheng-Han; Chang, Hsin-Yu; Tsai, Sheng-Feng; Lin, Tzu-Wei; Chen, Shean-Jen

    2016-01-01

    Temporal focusing multiphoton microscopy (TFMPM) has the advantage of area excitation in an axial confinement of only a few microns; hence, it can offer fast three-dimensional (3D) multiphoton imaging. Herein, fast volumetric imaging via a developed digital micromirror device (DMD)-based TFMPM has been realized through the synchronization of an electron multiplying charge-coupled device (EMCCD) with a dynamic piezoelectric stage for axial scanning. The volumetric imaging rate can achieve 30 volumes per second according to the EMCCD frame rate of more than 400 frames per second, which allows for the 3D Brownian motion of one-micron fluorescent beads to be spatially observed. Furthermore, it is demonstrated that the dynamic HiLo structural multiphoton microscope can reject background noise by way of the fast volumetric imaging with high-speed DMD patterned illumination. PMID:27231617

  11. Fast volumetric imaging with patterned illumination via digital micro-mirror device-based temporal focusing multiphoton microscopy.

    PubMed

    Chang, Chia-Yuan; Hu, Yvonne Yuling; Lin, Chun-Yu; Lin, Cheng-Han; Chang, Hsin-Yu; Tsai, Sheng-Feng; Lin, Tzu-Wei; Chen, Shean-Jen

    2016-05-01

    Temporal focusing multiphoton microscopy (TFMPM) has the advantage of area excitation in an axial confinement of only a few microns; hence, it can offer fast three-dimensional (3D) multiphoton imaging. Herein, fast volumetric imaging via a developed digital micromirror device (DMD)-based TFMPM has been realized through the synchronization of an electron multiplying charge-coupled device (EMCCD) with a dynamic piezoelectric stage for axial scanning. The volumetric imaging rate can achieve 30 volumes per second according to the EMCCD frame rate of more than 400 frames per second, which allows for the 3D Brownian motion of one-micron fluorescent beads to be spatially observed. Furthermore, it is demonstrated that the dynamic HiLo structural multiphoton microscope can reject background noise by way of the fast volumetric imaging with high-speed DMD patterned illumination. PMID:27231617

  12. Fast volumetric imaging with patterned illumination via digital micro-mirror device-based temporal focusing multiphoton microscopy.

    PubMed

    Chang, Chia-Yuan; Hu, Yvonne Yuling; Lin, Chun-Yu; Lin, Cheng-Han; Chang, Hsin-Yu; Tsai, Sheng-Feng; Lin, Tzu-Wei; Chen, Shean-Jen

    2016-05-01

    Temporal focusing multiphoton microscopy (TFMPM) has the advantage of area excitation in an axial confinement of only a few microns; hence, it can offer fast three-dimensional (3D) multiphoton imaging. Herein, fast volumetric imaging via a developed digital micromirror device (DMD)-based TFMPM has been realized through the synchronization of an electron multiplying charge-coupled device (EMCCD) with a dynamic piezoelectric stage for axial scanning. The volumetric imaging rate can achieve 30 volumes per second according to the EMCCD frame rate of more than 400 frames per second, which allows for the 3D Brownian motion of one-micron fluorescent beads to be spatially observed. Furthermore, it is demonstrated that the dynamic HiLo structural multiphoton microscope can reject background noise by way of the fast volumetric imaging with high-speed DMD patterned illumination.

  13. Multiphoton fluorescence lifetime imaging shows spatial segregation of secondary metabolites in Eucalyptus secretory cavities.

    PubMed

    Heskes, A M; Lincoln, C N; Goodger, J Q D; Woodrow, I E; Smith, T A

    2012-07-01

    Multiphoton fluorescence lifetime imaging provides an excellent tool for imaging deep within plant tissues while providing a means to distinguish between fluorophores with high spatial and temporal resolution. Ideal candidates for the application of multiphoton fluorescence lifetime imaging to plants are the embedded secretory cavities found in numerous species because they house complex mixtures of secondary metabolites within extracellular lumina. Previous investigations of this type of structure have been restricted by the use of sectioned material resulting in the loss of lumen contents and often disorganization of the delicate secretory cells; thus it is not known if there is spatial segregation of secondary metabolites within these structures. In this paper, we apply multiphoton fluorescence lifetime imaging to investigate the spatial arrangement of metabolites within intact secretory cavities isolated from Eucalyptus polybractea R.T. Baker leaves. The secretory cavities of this species are abundant (up to 10 000 per leaf), large (up to 6 nL) and importantly house volatile essential oil rich in the monoterpene 1,8-cineole, together with an immiscible, non-volatile component comprised largely of autofluorescent oleuropeic acid glucose esters. We have been able to optically section into the lumina of secretory cavities to a depth of ∼80 μm, revealing a unique spatial organization of cavity metabolites whereby the non-volatile component forms a layer between the secretory cells lining the lumen and the essential oil. This finding could be indicative of a functional role of the non-volatile component in providing a protective region of low diffusivity between the secretory cells and potentially autotoxic essential oil.

  14. Multiphoton imaging of excised normal skin and keloid scar: preliminary investigations

    NASA Astrophysics Data System (ADS)

    Brewer, Michael B.; Yeh, Alvin T.; Torkian, Behrooz; Sun, Chung-Ho; Tromberg, Bruce J.; Wong, Brian J.

    2004-07-01

    Wound healing is a physiologic process that acts to repair disruptions in the continuity of tissue caused by injury or surgical incision. Keloids and hypertrophic scars are forms of aberrant wound healing, which are characterized by the overproduction of collagen, resulting in an excessive amount of scar tissue. Keloid tumors, by definition, grow outside the boundary of the original tissue damage. Multiphoton microscopy (MPM) is an imaging technique which allows imaging of living specimens, without the use of fixation or stains. Images of collagen fibers are produced by the second harmonic signal intensity generated by endogenous fluorescence through excitation by infrared laser light. A postauricular keloid tumor was excised from a patient. The tissue was dissected, and a portion was imaged using MPM. Normal skin tissue was isolated from a patient undergoing a facelift. A portion of this tissue was also dissected and imaged using MPM. MPM images were taken using a 63X water immersion objective lens on a two-photon microscope and a titanium-sapphire laser. Images were taken beginning at the surface of the tissue and moving in at intervals of 200 nm to a final depth of 30 μm. The two-photon images were used to reconstruct three-dimensional representations of the collagen matrix within the tissues, which are readily contrasted. Density of the collagen within each tissue was also ascertained using depth dependant decay of the image intensity. Multiphoton imaging was successfully used to image the collagen matrix of normal skin and a keloid scar, demonstrating differences in their microstructures.

  15. Multimodal In Vivo Skin Imaging with Integrated Optical Coherence and Multiphoton Microscopy

    PubMed Central

    Graf, Benedikt W.; Boppart, Stephen A.

    2014-01-01

    In this paper, we demonstrate high-resolution, multimodal in vivo imaging of human skin using optical coherence (OCM) and multiphoton microscopy (MPM). These two modalities are integrated into a single instrument to enable simultaneous acquisition and coregistration. The system design and the OCM image processing architecture enable sufficient performance of both modalities for in vivo imaging of human skin. Examples of multimodal in vivo imaging are presented as well as time lapse imaging of blood flow in single capillary loops. By making use of multiple intrinsic contrast mechanisms this integrated technique improves the ability to noninvasively visualize living tissue. Integrated OCM and MPM has potential applications for in vivo diagnosis of various pathological skin conditions, such as skin cancer, as well as potential pharmaceutical and cosmetic research applications. PMID:25673966

  16. Characterization of multiphoton photoacoustic spectroscopy for subsurface brain tissue diagnosis and imaging

    NASA Astrophysics Data System (ADS)

    Dahal, Sudhir; Cullum, Brian M.

    2016-04-01

    The development and demonstration of a multiphoton photoacoustic imaging technique capable of providing high spatial resolution chemical images of subsurface tissue components as deep as 1.4 cm below the tissue surface is described. By combining multiphoton excitation in the diagnostic window (650 to 1100 nm), with ultrasonic detection of nonradiative relaxation events, it is possible to rapidly reconstruct three-dimensional, chemical specific, images of samples underneath overlying structures as well as chemical species of the same material. Demonstration of this technique for subsurface tissue differentiation is shown, with the ability to distinguish between grade III astrocytoma tissue and adjacent healthy tissue in blind studies. By employing photoacoustic signal detection, the high nonradiative relaxation rates of most biological tissue components (>90%) and the minimal signal attenuation of the resulting ultrasound compensate for excitation efficiency losses associated with two-photon absorption. Furthermore, the two-photon absorption process results in a highly localized excitation volume (ca., 60 μm). Characterization of the probing depth, spatial resolution, and ability to image through overlying structures is also demonstrated in this paper using tissue phantoms with well-characterized optical scattering properties, mimicking those of tissues.

  17. Characterization of multiphoton photoacoustic spectroscopy for subsurface brain tissue diagnosis and imaging

    NASA Astrophysics Data System (ADS)

    Dahal, Sudhir; Cullum, Brian M.

    2016-04-01

    The development and demonstration of a multiphoton photoacoustic imaging technique capable of providing high spatial resolution chemical images of subsurface tissue components as deep as 1.4 cm below the tissue surface is described. By combining multiphoton excitation in the diagnostic window (650 to 1100 nm), with ultrasonic detection of nonradiative relaxation events, it is possible to rapidly reconstruct three-dimensional, chemical specific, images of samples underneath overlying structures as well as chemical species of the same material. Demonstration of this technique for subsurface tissue differentiation is shown, with the ability to distinguish between grade III astrocytoma tissue and adjacent healthy tissue in blind studies. By employing photoacoustic signal detection, the high nonradiative relaxation rates of most biological tissue components (>90%) and the minimal signal attenuation of the resulting ultrasound compensate for excitation efficiency losses associated with two-photon absorption. Furthermore, the two-photon absorption process results in a highly localized excitation volume (ca., 60 μm). Characterization of the probing depth, spatial resolution, and ability to image through overlying structures is also demonstrated in this paper using tissue phantoms with well-characterized optical scattering properties, mimicking those of tissues.

  18. Optical workstation with concurrent, independent multiphoton imaging and experimental laser microbeam capabilities

    PubMed Central

    Wokosin, David L.; Squirrell, Jayne M.; Eliceiri, Kevin W.; White, John G.

    2008-01-01

    Experimental laser microbeam techniques have become established tools for studying living specimens. A steerable, focused laser beam may be used for a variety of experimental manipulations such as laser microsurgery, optical trapping, localized photolysis of caged bioactive probes, and patterned photobleaching. Typically, purpose-designed experimental systems have been constructed for each of these applications. In order to assess the consequences of such experimental optical interventions, long-term, microscopic observation of the specimen is often required. Multiphoton excitation, because of its ability to obtain high-contrast images from deep within a specimen with minimal phototoxic effects, is a preferred technique for in vivo imaging. An optical workstation is described that combines the functionality of an experimental optical microbeam apparatus with a sensitive multiphoton imaging system designed for use with living specimens. Design considerations are discussed and examples of ongoing biological applications are presented. The integrated optical workstation concept offers advantages in terms of flexibility and versatility relative to systems implemented with separate imaging and experimental components. PMID:18607511

  19. Characterization of multiphoton laser scanning device optical parameters for image restoration

    NASA Astrophysics Data System (ADS)

    Fischer, Frank; Konig, Karsten; Puschmann, Stefan; Wepf, Roger; Riemann, Iris; Ulrich, Volker; Fischer, Peter

    2004-09-01

    Fluorescent nanobeads embedded in agarose and skin biopsies were used to optically characterize spatial and temporal resolution of multiphoton laser scanning devices (MPLSD). Optical sections based on two-photon excited bead fluorescence have been performed at various sample depths. Three-dimensional reconstruction of the image stacks allowed determination of the point spread function. Using calculated point spread functions to apply deconvolution procedures (e.g. Huygens software), the visualization and hence the interpretation of intradermal structures, such as extracellular matrix components in 150 μm tissue depth, was improved.

  20. Imaging sulfur mustard lesions in human epidermal tissues and keratinocytes by confocal and multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Werrlein, Robert; Madren-Whalley, Janna S.

    2002-06-01

    Topical exposure to sulfur mustard (HD), a known theat agent, produces persistent and debilitating cutaneous blisters. The blisters occur at the dermal-epidermal junction following a dose-dependent latent period of 8-24 h, however, the primary lesions causing vesication remain uncertain. Immunofluorescent images reveal that a 5-min exposure to 400 (mu) M HD disrupts molecules that are also disrupted by epidermolysis bullosa-type blistering diseases of the skin. Using keratinocyte cultures and fluorochomes conjugated to two different keratin-14 (K14) antibodies (clones CKB1 and LL002), results have shown a statistically significant (p<0.1) 1-h decrease of 29.2% in expression of the CKB1 epitope, a nearly complete loss of CKB1 expression within 2 h, and progressive cytoskeletal (K14) collapse without loss in expression of the LL002 epitope. With human epidermal tissues, multi-photon images of (alpha) 6 integrin and laminin 5 showed disruptive changes in the cell-surface organization and integrity of these adhesion molecules. At 1 H postexposure, analyses showed a statistically significant (p<0.1) decrease of 27.3% in (alpha) 6 integrin emissions, and a 32% decrease in laminin 5 volume. Multi-photon imaging indicates that molecules essential for epidermal-dermal attachment are early targets in the alkylating events leading to HD-induced vesication.

  1. In vivo imaging of spinal cord in contusion injury model mice by multi-photon microscopy

    NASA Astrophysics Data System (ADS)

    Oshima, Y.; Horiuchi, H.; Ogata, T.; Hikita, A.; Miura, H.; Imamura, T.

    2014-03-01

    Fluorescent imaging technique is a promising method and has been developed for in vivo applications in cellular biology. In particular, nonlinear optical imaging technique, multi-photon microscopy has make it possible to analyze deep portion of tissues in living animals such as axons of spinal code. Traumatic spinal cord injuries (SCIs) are usually caused by contusion damages. Therefore, observation of spinal cord tissue after the contusion injury is necessary for understanding cellular dynamics in response to traumatic SCI and development of the treatment for traumatic SCI. Our goal is elucidation of mechanism for degeneration of axons after contusion injuries by establishing SCI model and chronic observation of injured axons in the living animals. Firstly we generated and observed acute SCI model by contusion injury. By using a multi-photon microscope, axons in dorsal cord were visualized approximately 140 micron in depth from the surface. Immediately after injury, minimal morphological change of spinal cord was observed. At 3 days after injury, spinal cord was swelling and the axons seem to be fragmented. At 7 days after injury, increased degradation of axons could be observed, although the image was blurred due to accumulation of the connective tissue. In the present study, we successfully observed axon degeneration after the contusion SCI in a living animal in vivo. Our final goal is to understand molecular mechanisms and cellular dynamics in response to traumatic SCIs in acute and chronic stage.

  2. Setup and use of a two-laser multiphoton microscope for multichannel intravital fluorescence imaging

    PubMed Central

    Entenberg, David; Wyckoff, Jeffrey; Gligorijevic, Bojana; Roussos, Evanthia T; Verkhusha, Vladislav V; Pollard, Jeffrey W; Condeelis, John

    2014-01-01

    Characterizing biological mechanisms dependent upon the interaction of many cell types in vivo requires both multiphoton microscope systems capable of expanding the number and types of fluorophores that can be imaged simultaneously while removing the wavelength and tunability restrictions of existing systems, and enhanced software for extracting critical cellular parameters from voluminous 4D data sets. We present a procedure for constructing a two-laser multiphoton microscope that extends the wavelength range of excitation light, expands the number of simultaneously usable fluorophores and markedly increases signal to noise via ‘over-clocking’ of detection. We also utilize a custom-written software plug-in that simplifies the quantitative tracking and analysis of 4D intravital image data. We begin by describing the optics, hardware, electronics and software required, and finally the use of the plug-in for analysis. We demonstrate the use of the setup and plug-in by presenting data collected via intravital imaging of a mouse model of breast cancer. The procedure may be completed in ~24 h. PMID:21959234

  3. Chronic imaging of amyloid plaques in the live mouse brain using multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Bacskai, Brian J.; Kajdasz, Stephen T.; Christie, R. H.; Zipfel, Warren R.; Williams, Rebecca M.; Kasischke, Karl A.; Webb, Watt W.; Hyman, B. T.

    2001-04-01

    Transgenic mice expressing the human Amyloid Precursor Protein (APP) develop amyloid plaques as they age. These plaques resemble those found in the human disease. Multiphoton laser scanning microscopy combined with a novel surgical approach was used to measure amyloid plaque dynamics chronically in the cortex of living transgenic mice. Thioflavine S (thioS) was used as a fluorescent marker of amyloid deposits. Multiphoton excitation allowed visualization of amyloid plaques up to 200 micrometers deep into the brain. The surgical site could be imaged repeatedly without overt damage to the tissue, and individual plaques within this volume could be reliably identified over periods of several days to several months. On average, plaque sizes remained constant over time, supporting a model of rapid deposition, followed by relative stability. Alternative reporters for in vivo histology include thiazine red, and FITC-labeled amyloid-(Beta) peptide. We also present examples of multi-color imaging using Hoechst dyes and FITC-labeled tomato lectin. These approaches allow us to observe cell nuclei or microglia simultaneously with amyloid-(Beta) deposits in vivo. Chronic imaging of a variety of reporters in these transgenic mice should provide insight into the dynamics of amyloid-(Beta) activity in the brain.

  4. Time-resolved multiphoton imaging of basal cell carcinoma

    NASA Astrophysics Data System (ADS)

    Cicchi, R.; Sestini, S.; De Giorgi, V.; Stambouli, D.; Carli, P.; Massi, D.; Pavone, F. S.

    2007-02-01

    We investigated human cutaneous basal cell carcinoma ex-vivo samples by combined time resolved two photon intrinsic fluorescence and second harmonic generation microscopy. Morphological and spectroscopic differences were found between malignant skin and corresponding healthy skin tissues. In comparison with normal healthy skin, cancer tissue showed a different morphology and a mean fluorescence lifetime distribution slightly shifted towards higher values. Topical application of delta-aminolevulinic acid to the lesion four hours before excision resulted in an enhancement of the fluorescence signal arising from malignant tissue, due to the accumulation of protoporphyrines inside tumor cells. Contrast enhancement was prevalent at tumor borders by both two photon fluorescence microscopy and fluorescence lifetime imaging. Fluorescence-based images showed a good correlation with conventional histopathological analysis, thereby supporting the diagnostic accuracy of this novel method. Combined morphological and lifetime analysis in the study of ex-vivo skin samples discriminated benign from malignant tissues, thus offering a reliable, non-invasive tool for the in-vivo analysis of inflammatory and neoplastic skin lesions.

  5. Extracting quantitative parameters from images in multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Zimmerley, Maxwell Stuart

    Coherent anti-Stokes Raman scattering (CARS) microscopy allows for fast, three-dimensionally resolved detection of molecules based on vibrational contrast. In CARS, the generated signal is nonlinearly dependent upon the concentration of the vibrational mode of interest. This makes it challenging to extract quantitative parameters (such as the concentration or orientation) from CARS images of biological and synthetic samples. Because of this, many investigations which employ CARS microscopy generally only report qualitative information extracted from these images. In this thesis, three methods have been developed to extract the quantitative concentration information from CARS images. In the first, the ratio of the forward-propagating and back-reflected CARS signal generated in tissue is used to monitor the percolation of DMSO into excised human cadaver skin. Through this, we find that the maximum clearing of skin with DMSO occurs at 40% v/v. We also combine CARS with second harmonic generation (SHG) to investigate the effects of DMSO on collagen. Up to a 20% v/v concentration of DMSO in the skin, the collagen becomes disrupted, resulting in a significant drop in the generated SHG. In the second method, the ratio between the CARS resonance peak and dip is correlated with the concentration to measure the concentration of water and deuterated glycine in hair. Both molecules are found to distribute throughout the hair fiber homogenously, water at a 34% v/v concentration, and d-glycine with a 0.22 M concentration. In the final method, CARS spectra over one vibrational mode are used to extract the imaginary part of the third-order nonlinear susceptibility. This quantity is linearly dependent upon the concentration of the vibrational mode of interest. This procedure is used to determine the degree of conversion of two-photon polymerized microstructures synthesized with varying writing powers. A sigmoidal relationship is observed between the applied intensity and the degree

  6. Hybrid Multiphoton Volumetric Functional Imaging of Large Scale Bioengineered Neuronal Networks

    PubMed Central

    Paluch, Shir; Dvorkin, Roman; Brosh, Inbar; Shoham, Shy

    2014-01-01

    Planar neural networks and interfaces serve as versatile in vitro models of central nervous system physiology, but adaptations of related methods to three dimensions (3D) have met with limited success. Here, we demonstrate for the first time volumetric functional imaging in a bio-engineered neural tissue growing in a transparent hydrogel with cortical cellular and synaptic densities, by introducing complementary new developments in nonlinear microscopy and neural tissue engineering. Our system uses a novel hybrid multiphoton microscope design combining a 3D scanning-line temporal-focusing subsystem and a conventional laser-scanning multiphoton microscope to provide functional and structural volumetric imaging capabilities: dense microscopic 3D sampling at tens of volumes/sec of structures with mm-scale dimensions containing a network of over 1000 developing cells with complex spontaneous activity patterns. These developments open new opportunities for large-scale neuronal interfacing and for applications of 3D engineered networks ranging from basic neuroscience to the screening of neuroactive substances. PMID:24898000

  7. Multiphoton microscopic imaging of fibrotic focus in invasive ductal carcinoma of the breast

    NASA Astrophysics Data System (ADS)

    Chen, Sijia; Nie, Yuting; Lian, Yuane; Wu, Yan; Fu, Fangmeng; Wang, Chuan; Zhuo, Shuangmu; Chen, Jianxin

    2014-11-01

    During the proliferation of breast cancer, the desmoplastic can evoke a fibrosis response by invading healthy tissue. Fibrotic focus (FF) in invasive ductal carcinoma (IDC) of the breast had been reported to be associated with significantly poorer survival rate than IDC without FF. As an important prognosis indicator, it's difficult to obtain the exact fibrotic information from traditional detection method such as mammography. Multiphoton imaging based on two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) has been recently employed for microscopic examination of unstained tissue. In this study, multiphoton microscopy (MPM) was used to image the fibrotic focus in invasive ductal carcinoma tissue. The morphology and distribution of collagen in fibrotic focus can be demonstrated by the SHG signal. Variation of collagen between IDC with and without FF will be examined and further characterized, which may be greatly related to the metastasis of breast cancer. Our result suggested that the MPM can be efficient in identifying and locating the fibrotic focus in IDC. Combining with the pathology analysis and other detecting methods, MPM owns potential in becoming an advanced histological tool for detecting the fibrotic focus in IDC and collecting prognosis information, which may guide the subsequent surgery option and therapy procedure for patients.

  8. Role of multiphoton bunching in high-order ghost imaging with thermal light sources

    SciTech Connect

    Liu Qian; Chen Xihao; Luo Kaihong; Wu Lingan; Wu Wei

    2009-05-15

    The intrinsic higher-order correlation of intensities which gives a measure of 'pure' correlations among photons (corresponding to multiphoton bunching) is investigated with regard to ghost imaging with thermal light. The synchronous detection of the same light field by all reference detectors, which is a necessary condition for achieving an Nth-order ghost image based on N-photon bunching, is discussed. Furthermore, it is found that the enhanced high visibility of Nth-order ghost imaging is a consequence of the contribution of N-photon bunching, which is not a small value but is equal to the sum of all contributions from (N-1)-photon bunching. These results differ from those obtained by certain other groups.

  9. Label-free in vivo imaging of Drosophila melanogaster by multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Lin, Chiao-Ying; Hovhannisyan, Vladimir; Wu, June-Tai; Lin, Sung-Jan; Lin, Chii-Wann; Chen, Jyh-Horng; Dong, Chen-Yuan

    2008-02-01

    The fruit fly Drosophila melanogaster is one of the most valuable organisms in genetic and developmental biology studies. Drosophila is a small organism with a short life cycle, and is inexpensive and easy to maintain. The entire genome of Drosophila has recently been sequenced (cite the reference). These advantages make fruit fly an attractive model organism for biomedical researches. Unlike humans, Drosophila can be subjected to genetic manipulation with relative ease. Originally, Drosophila was mostly used in classical genetics studies. In the model era of molecular biology, the fruit fly has become a model organ for developmental biology researches. In the past, numerous molecularly modified mutants with well defined genetic defects affecting different aspects of the developmental processes have been identified and studied. However, traditionally, the developmental defects of the mutant flies are mostly examined in isolated fixed tissues which preclude the observation of the dynamic interaction of the different cell types and the extracellular matrix. Therefore, the ability to image different organelles of the fruit fly without extrinsic labeling is invaluable for Drosophila biology. In this work, we successfully acquire in vivo images of both developing muscles and axons of motor neurons in the three larval stages by using the minimially invasive imaging modality of multiphoton (SHG) microscopy. We found that while SHG imaging is useful in revealing the muscular architecture of the developing larva, it is the autofluorescence signal that allows label-free imaging of various organelles to be achieved. Our results demonstrate that multiphoton imaging is a powerful technique for investigation the development of Drosophila.

  10. Heuristically optimal path scanning for high-speed multiphoton circuit imaging.

    PubMed

    Sadovsky, Alexander J; Kruskal, Peter B; Kimmel, Joseph M; Ostmeyer, Jared; Neubauer, Florian B; MacLean, Jason N

    2011-09-01

    Population dynamics of patterned neuronal firing are fundamental to information processing in the brain. Multiphoton microscopy in combination with calcium indicator dyes allows circuit dynamics to be imaged with single-neuron resolution. However, the temporal resolution of fluorescent measures is constrained by the imaging frequency imposed by standard raster scanning techniques. As a result, traditional raster scans limit the ability to detect the relative timing of action potentials in the imaged neuronal population. To maximize the speed of fluorescence measures from large populations of neurons using a standard multiphoton laser scanning microscope (MPLSM) setup, we have developed heuristically optimal path scanning (HOPS). HOPS optimizes the laser travel path length, and thus the temporal resolution of neuronal fluorescent measures, using standard galvanometer scan mirrors. Minimizing the scan path alone is insufficient for prolonged high-speed imaging of neuronal populations. Path stability and the signal-to-noise ratio become increasingly important factors as scan rates increase. HOPS addresses this by characterizing the scan mirror galvanometers to achieve prolonged path stability. In addition, the neuronal dwell time is optimized to sharpen the detection of action potentials while maximizing scan rate. The combination of shortest path calculation and minimization of mirror positioning time allows us to optically monitor a population of neurons in a field of view at high rates with single-spike resolution, ∼ 125 Hz for 50 neurons and ∼ 8.5 Hz for 1,000 neurons. Our approach introduces an accessible method for rapid imaging of large neuronal populations using traditional MPLSMs, facilitating new insights into neuronal circuit dynamics.

  11. Heuristically optimal path scanning for high-speed multiphoton circuit imaging.

    PubMed

    Sadovsky, Alexander J; Kruskal, Peter B; Kimmel, Joseph M; Ostmeyer, Jared; Neubauer, Florian B; MacLean, Jason N

    2011-09-01

    Population dynamics of patterned neuronal firing are fundamental to information processing in the brain. Multiphoton microscopy in combination with calcium indicator dyes allows circuit dynamics to be imaged with single-neuron resolution. However, the temporal resolution of fluorescent measures is constrained by the imaging frequency imposed by standard raster scanning techniques. As a result, traditional raster scans limit the ability to detect the relative timing of action potentials in the imaged neuronal population. To maximize the speed of fluorescence measures from large populations of neurons using a standard multiphoton laser scanning microscope (MPLSM) setup, we have developed heuristically optimal path scanning (HOPS). HOPS optimizes the laser travel path length, and thus the temporal resolution of neuronal fluorescent measures, using standard galvanometer scan mirrors. Minimizing the scan path alone is insufficient for prolonged high-speed imaging of neuronal populations. Path stability and the signal-to-noise ratio become increasingly important factors as scan rates increase. HOPS addresses this by characterizing the scan mirror galvanometers to achieve prolonged path stability. In addition, the neuronal dwell time is optimized to sharpen the detection of action potentials while maximizing scan rate. The combination of shortest path calculation and minimization of mirror positioning time allows us to optically monitor a population of neurons in a field of view at high rates with single-spike resolution, ∼ 125 Hz for 50 neurons and ∼ 8.5 Hz for 1,000 neurons. Our approach introduces an accessible method for rapid imaging of large neuronal populations using traditional MPLSMs, facilitating new insights into neuronal circuit dynamics. PMID:21715667

  12. In vivo multiphoton imaging of human skin: assessment of topical corticosteroid-induced epidermis atrophy and depigmentation

    NASA Astrophysics Data System (ADS)

    Ait El Madani, Hassan; Tancrède-Bohin, Emmanuelle; Bensussan, Armand; Colonna, Anne; Dupuy, Alain; Bagot, Martine; Pena, Ana-Maria

    2012-02-01

    Multiphoton microscopy has emerged in the past decade as a promising tool for noninvasive skin imaging. Our aim was to evaluate the potential of multiphoton microscopy to detect topical corticosteroids side effects within the epidermis and to provide new insights into their dynamics. Healthy volunteers were topically treated with clobetasol propionate on a small region of their forearms under overnight occlusion for three weeks. The treated region of each patient was investigated at D0, D7, D15, D22 (end of the treatment), and D60. Our study shows that multiphoton microscopy allows for the detection of corticoid-induced epidermis modifications: thinning of stratum corneum compactum and epidermis, decrease of keratinocytes size, and changes in their morphology from D7 to D22. We also show that multiphoton microscopy enables in vivo three-dimensional (3-D) quantitative assessment of melanin content. We observe that melanin density decreases during treatment and almost completely disappears at D22. Moreover, these alterations are reversible as they are no longer present at D60. Our study demonstrates that multiphoton microscopy is a convenient and powerful tool for noninvasive 3-D dynamical studies of skin integrity and pigmentation.

  13. Automatic centerline extraction of irregular tubular structures using probability volumes from multiphoton imaging.

    PubMed

    Santamaría-Pang, A; Colbert, C M; Saggau, P; Kakadiaris, I A

    2007-01-01

    In this paper, we present a general framework for extracting 3D centerlines from volumetric datasets. Unlike the majority of previous approaches, we do not require a prior segmentation of the volume nor we do assume any particular tubular shape. Centerline extraction is performed using a morphology-guided level set model. Our approach consists of: i) learning the structural patterns of a tubular-like object, and ii) estimating the centerline of a tubular object as the path with minimal cost with respect to outward flux in gray level images. Such shortest path is found by solving the Eikonal equation. We compare the performance of our method with existing approaches in synthetic, CT, and multiphoton 3D images, obtaining substantial improvements, especially in the case of irregular tubular objects. PMID:18044604

  14. Imaging photoelectron circular dichroism of chiral molecules by femtosecond multiphoton coincidence detection

    NASA Astrophysics Data System (ADS)

    Lehmann, C. Stefan; Ram, N. Bhargava; Powis, Ivan; Janssen, Maurice H. M.

    2013-12-01

    Here, we provide a detailed account of novel experiments employing electron-ion coincidence imaging to discriminate chiral molecules. The full three-dimensional angular scattering distribution of electrons is measured after photoexcitation with either left or right circular polarized light. The experiment is performed using a simplified photoelectron-photoion coincidence imaging setup employing only a single particle imaging detector. Results are reported applying this technique to enantiomers of the chiral molecule camphor after three-photon ionization by circularly polarized femtosecond laser pulses at 400 nm and 380 nm. The electron-ion coincidence imaging provides the photoelectron spectrum of mass-selected ions that are observed in the time-of-flight mass spectra. The coincident photoelectron spectra of the parent camphor ion and the various fragment ions are the same, so it can be concluded that fragmentation of camphor happens after ionization. We discuss the forward-backward asymmetry in the photoelectron angular distribution which is expressed in Legendre polynomials with moments up to order six. Furthermore, we present a method, similar to one-photon electron circular dichroism, to quantify the strength of the chiral electron asymmetry in a single parameter. The circular dichroism in the photoelectron angular distribution of camphor is measured to be 8% at 400 nm. The electron circular dichroism using femtosecond multiphoton excitation is of opposite sign and about 60% larger than the electron dichroism observed before in near-threshold one-photon ionization with synchrotron excitation. We interpret our multiphoton ionization as being resonant at the two-photon level with the 3s and 3p Rydberg states of camphor. Theoretical calculations are presented that model the photoelectron angular distribution from a prealigned camphor molecule using density functional theory and continuum multiple scattering X alpha photoelectron scattering calculations

  15. Imaging photoelectron circular dichroism of chiral molecules by femtosecond multiphoton coincidence detection

    SciTech Connect

    Lehmann, C. Stefan; Ram, N. Bhargava; Janssen, Maurice H. M.; Powis, Ivan

    2013-12-21

    Here, we provide a detailed account of novel experiments employing electron-ion coincidence imaging to discriminate chiral molecules. The full three-dimensional angular scattering distribution of electrons is measured after photoexcitation with either left or right circular polarized light. The experiment is performed using a simplified photoelectron-photoion coincidence imaging setup employing only a single particle imaging detector. Results are reported applying this technique to enantiomers of the chiral molecule camphor after three-photon ionization by circularly polarized femtosecond laser pulses at 400 nm and 380 nm. The electron-ion coincidence imaging provides the photoelectron spectrum of mass-selected ions that are observed in the time-of-flight mass spectra. The coincident photoelectron spectra of the parent camphor ion and the various fragment ions are the same, so it can be concluded that fragmentation of camphor happens after ionization. We discuss the forward-backward asymmetry in the photoelectron angular distribution which is expressed in Legendre polynomials with moments up to order six. Furthermore, we present a method, similar to one-photon electron circular dichroism, to quantify the strength of the chiral electron asymmetry in a single parameter. The circular dichroism in the photoelectron angular distribution of camphor is measured to be 8% at 400 nm. The electron circular dichroism using femtosecond multiphoton excitation is of opposite sign and about 60% larger than the electron dichroism observed before in near-threshold one-photon ionization with synchrotron excitation. We interpret our multiphoton ionization as being resonant at the two-photon level with the 3s and 3p Rydberg states of camphor. Theoretical calculations are presented that model the photoelectron angular distribution from a prealigned camphor molecule using density functional theory and continuum multiple scattering X alpha photoelectron scattering calculations

  16. Spectral image reconstruction through the PCA transform

    NASA Astrophysics Data System (ADS)

    Ma, Long; Qiu, Xuewei; Cong, Yangming

    2015-12-01

    Digital color image reproduction based on spectral information has become a field of much interest and practical importance in recent years. The representation of color in digital form with multi-band images is not very accurate, hence the use of spectral image is justified. Reconstructing high-dimensional spectral reflectance images from relatively low-dimensional camera signals is generally an ill-posed problem. The aim of this study is to use the Principal component analysis (PCA) transform in spectral reflectance images reconstruction. The performance is evaluated by the mean, median and standard deviation of color difference values. The values of mean, median and standard deviation of root mean square (GFC) errors between the reconstructed and the actual spectral image were also calculated. Simulation experiments conducted on a six-channel camera system and on spectral test images show the performance of the suggested method.

  17. New developments in multimodal clinical multiphoton tomography

    NASA Astrophysics Data System (ADS)

    König, Karsten

    2011-03-01

    80 years ago, the PhD student Maria Goeppert predicted in her thesis in Goettingen, Germany, two-photon effects. It took 30 years to prove her theory, and another three decades to realize the first two-photon microscope. With the beginning of this millennium, first clinical multiphoton tomographs started operation in research institutions, hospitals, and in the cosmetic industry. The multiphoton tomograph MPTflexTM with its miniaturized flexible scan head became the Prism-Award 2010 winner in the category Life Sciences. Multiphoton tomographs with its superior submicron spatial resolution can be upgraded to 5D imaging tools by adding spectral time-correlated single photon counting units. Furthermore, multimodal hybrid tomographs provide chemical fingerprinting and fast wide-field imaging. The world's first clinical CARS studies have been performed with a hybrid multimodal multiphoton tomograph in spring 2010. In particular, nonfluorescent lipids and water as well as mitochondrial fluorescent NAD(P)H, fluorescent elastin, keratin, and melanin as well as SHG-active collagen have been imaged in patients with dermatological disorders. Further multimodal approaches include the combination of multiphoton tomographs with low-resolution imaging tools such as ultrasound, optoacoustic, OCT, and dermoscopy systems. Multiphoton tomographs are currently employed in Australia, Japan, the US, and in several European countries for early diagnosis of skin cancer (malignant melanoma), optimization of treatment strategies (wound healing, dermatitis), and cosmetic research including long-term biosafety tests of ZnO sunscreen nanoparticles and the measurement of the stimulated biosynthesis of collagen by anti-ageing products.

  18. Staring 2-D hadamard transform spectral imager

    DOEpatents

    Gentry, Stephen M.; Wehlburg, Christine M.; Wehlburg, Joseph C.; Smith, Mark W.; Smith, Jody L.

    2006-02-07

    A staring imaging system inputs a 2D spatial image containing multi-frequency spectral information. This image is encoded in one dimension of the image with a cyclic Hadamarid S-matrix. The resulting image is detecting with a spatial 2D detector; and a computer applies a Hadamard transform to recover the encoded image.

  19. Marginal characteristics of skin scarred dermis quantitatively extracted from multiphoton microscopic imaging

    NASA Astrophysics Data System (ADS)

    Zhu, Xiaoqin; Zhuo, Shuangmu; Zheng, Liqin; Jiang, Xingshan; Chen, Jianxin; Lin, Bifang

    2010-11-01

    Multiphoton microscopy based on two-photon excited fluorescence (TPEF) and second harmonic generation (SHG) was applied to examine the marginal regions at dermis of normal, atrophic and keloid scars. High-contrast, high-resolution image showed an obvious boundary at scar margin and different morphological patterns of collagen or elastin on the two sides. Since the degree of the morphological alteration between the two sides of boundary at scar margin was varied among different types of scars, alteration degree of SHG-to-TPEF index was defined as a quantitative indicator for discrimination. It will help to determine the most appropriate clinical treatment strategy for different types of scars and potentially monitor therapy in vivo.

  20. Label-free multi-photon imaging of dysplasia in Barrett’s esophagus

    PubMed Central

    Mehravar, Soroush; Banerjee, Bhaskar; Chatrath, Hemant; Amirsolaimani, Babak; Patel, Krunal; Patel, Charmi; Norwood, Robert A; Peyghambarian, Nasser; Kieu, Khanh

    2015-01-01

    Barrett’s esophagus (BE) is a metaplastic disorder where dysplastic and early cancerous changes are invisible to the naked eye and where the practice of blind biopsy is hampered by large sampling errors. Multi-photon microscopy (MPM) has emerged as an alternative solution for fast and label-free diagnostic capability for identifying the histological features with sub-micron accuracy. We developed a compact, inexpensive MPM system by using a handheld mode-locked fiber laser operating at 1560nm to study mucosal biopsies of BE. The combination of back-scattered THG, back-reflected forward THG and SHG signals generate images of cell nuclei and collagen, leading to label-free diagnosis in Barrett’s. PMID:26819824

  1. A database for spectral image quality

    NASA Astrophysics Data System (ADS)

    Le Moan, Steven; George, Sony; Pedersen, Marius; Blahová, Jana; Hardeberg, Jon Yngve

    2015-01-01

    We introduce a new image database dedicated to multi-/hyperspectral image quality assessment. A total of nine scenes representing pseudo-at surfaces of different materials (textile, wood, skin. . . ) were captured by means of a 160 band hyperspectral system with a spectral range between 410 and 1000nm. Five spectral distortions were designed, applied to the spectral images and subsequently compared in a psychometric experiment, in order to provide a basis for applications such as the evaluation of spectral image difference measures. The database can be downloaded freely from http://www.colourlab.no/cid.

  2. Simultaneous multiple-excitation multiphoton microscopy yields increased imaging sensitivity and specificity

    PubMed Central

    2011-01-01

    Background Multiphoton microscopy (MPM) offers many advantages over conventional wide-field and confocal laser scanning microscopy (CLSM) for imaging biological samples such as 3D resolution of excitation, reduced phototoxicity, and deeper tissue imaging. However, adapting MPM for critical multi-color measurements presents a challenge because of the largely overlapping two-photon absorption (TPA) peaks of common biological fluorophores. Currently, most multi-color MPM relies on the absorbance at one intermediate wavelength of multiple dyes, which introduces problems such as decreased and unequal excitation efficiency across the set of dyes. Results Here we describe an MPM system incorporating two, independently controlled sources of two-photon excitation whose wavelengths are adjusted to maximally excite one dye while minimally exciting the other. We report increased signal-to-noise ratios and decreased false positive emission bleed-through using this novel multiple-excitation MPM (ME-MPM) compared to conventional single-excitation MPM (SE-MPM) in a variety of multi-color imaging applications. Conclusions Similar to the tremendous gain in popularity of CLSM after the introduction of multi-color imaging, we anticipate that the ME-MPM system will further increase the popularity of MPM. In addition, ME-MPM provides an excellent tool to more rapidly design and optimize pairs of fluorescence probes for multi-color two-photon imaging, such as CFP/YFP or GFP/DsRed for CLSM. PMID:21366923

  3. Multiphoton microscopy based cryo-imaging of inflated frozen human lung sections at -60°C in healthy and COPD lungs

    NASA Astrophysics Data System (ADS)

    Abraham, Thomas; Kayra, Damian; Zhang, Angela; Suzuki, Masaru; McDonough, John; Elliott, W. M.; Cooper, Joel D.; Hogg, James C.

    2013-02-01

    Lung is a complex gas exchanger with interfacial area (where the gas exchange takes place) is about the size of a tennis court. Respiratory function is linked to the biomechanical stability of the gas exchange or alveolar regions which directly depends on the spatial distributions of the extracellular matrix fibers such fibrillar collagens and elastin fibers. It is very important to visualize and quantify these fibers at their native and inflated conditions to have correct morphometric information on differences between control and diseased states. This can be only achieved in the ex vivo states by imaging directly frozen lung specimens inflated to total lung capacity. Multiphoton microscopy, which uses ultra-short infrared laser pulses as the excitation source, produces multiphoton excitation fluorescence (MPEF) signals from endogenously fluorescent proteins (e.g. elastin) and induces specific second harmonic generation (SHG) signals from non-centrosymmetric proteins such as fibrillar collagens in fresh human lung tissues [J. Struct. Biol. (2010)171,189-196]. Here we report for the first time 3D image data obtained directly from thick frozen inflated lung specimens (~0.7- 1.0 millimeter thick) visualized at -60°C without prior fixation or staining in healthy and diseased states. Lung specimens donated for transplantation and released for research when no appropriate recipient was identified served as controls, and diseased lung specimens donated for research by patients receiving lung transplantation for very severe COPD (n=4) were prepared as previously described [N. Engl. J. Med. (2011) 201, 1567]. Lung slices evenly spaced between apex and base were examined using multiphoton microscopy while maintained at -60°C using a temperature controlled cold stage with a temperature resolution of 0.1°C. Infrared femto-second laser pulses tuned to 880nm, dry microscopic objectives, and non-de-scanned detectors/spectrophotometer located in the reflection geometry were

  4. Multiphoton microscopy as a diagnostic imaging modality for pancreatic neoplasms without hematoxylin and eosin stains.

    PubMed

    Chen, Youting; Chen, Jing; Chen, Hong; Hong, Zhipeng; Zhu, Xiaoqin; Zhuo, Shuangmu; Chen, Yanling; Chen, Jianxin

    2014-09-01

    Hematoxylin and eosin (H&E) staining of tissue samples is the standard approach in histopathology for imaging and diagnosing cancer. Recent reports have shown that multiphoton microscopy (MPM) provides better sample interface with single-cell resolution, which enhances traditional H&E staining and offers a powerful diagnostic tool with potential applications in oncology. The purpose of this study was to further expand the versatility of MPM by establishing the optical parameters required for imaging unstained histological sections of pancreatic neoplasms, thereby providing an efficient and environmentally sustainable alternative to H&E staining while improving the accuracy of pancreatic cancer diagnoses. We found that the high-resolution MPM images clearly distinguish between the structure of normal pancreatic tissues compared with pancreatic neoplasms in unstained histological sections, and discernable differences in tissue architecture and cell morphology between normal versus tumorigenic cells led to enhanced optical diagnosis of cancerous tissue. Moreover, quantitative assessment of the cytomorphological features visualized from MPM images showed significant differences in the nuclear–cytoplasmic ratios of pancreatic neoplasms compared with normal pancreas, as well as further distinguished pancreatic malignant tumors from benign tumors. These results indicate that the MPM could potentially serve as an optical tool for the diagnosis of pancreatic neoplasms in unstained histological sections.

  5. Does optical microangiography provide accurate imaging of capillary vessels?: validation using multiphoton microscopy

    PubMed Central

    Wang, Hequn; Baran, Utku; Li, Yuandong; Qin, Wan; Wang, Wenbo; Zeng, Haishan; Wang, Ruikang K.

    2014-01-01

    Abstract. Optical microangiography (OMAG) has been extensively utilized to study three-dimensional tissue vasculature in vivo. However, with the limited image resolution (∼10  μm) of the commonly used systems, some concerns were raised: (1) whether OMAG is capable of providing the imaging of capillary vessels that are of an average diameter of ∼6  μm; (2) if yes, whether OMAG can provide meaningful quantification of vascular density within the scanned tissue volume. Multiphoton microscopy (MPM) is capable of depth-resolved high-resolution (∼1  μm) imaging of biological tissue structures. With externally labeled plasma, the vascular network including single capillaries can be well visualized. We compare the vascular images of in vivo mouse brain acquired by both OMAG and MPM systems. We found that within the penetration depth range of the MPM system, OMAG is able to accurately visualize blood vessels including capillaries. Although the resolution of OMAG may not be able to 100% resolve two closely packed tiny capillaries in tissue, it is still capable of visualizing most of the capillaries because there are interstitial tissue spaces between them. We believe our validation results reinforce the application of OMAG in microvasculature-related studies. PMID:25341071

  6. Multiphoton microscopy as a diagnostic imaging modality for pancreatic neoplasms without hematoxylin and eosin stains

    NASA Astrophysics Data System (ADS)

    Chen, Youting; Chen, Jing; Chen, Hong; Hong, Zhipeng; Zhu, Xiaoqin; Zhuo, Shuangmu; Chen, Yanling; Chen, Jianxin

    2014-09-01

    Hematoxylin and eosin (H&E) staining of tissue samples is the standard approach in histopathology for imaging and diagnosing cancer. Recent reports have shown that multiphoton microscopy (MPM) provides better sample interface with single-cell resolution, which enhances traditional H&E staining and offers a powerful diagnostic tool with potential applications in oncology. The purpose of this study was to further expand the versatility of MPM by establishing the optical parameters required for imaging unstained histological sections of pancreatic neoplasms, thereby providing an efficient and environmentally sustainable alternative to H&E staining while improving the accuracy of pancreatic cancer diagnoses. We found that the high-resolution MPM images clearly distinguish between the structure of normal pancreatic tissues compared with pancreatic neoplasms in unstained histological sections, and discernable differences in tissue architecture and cell morphology between normal versus tumorigenic cells led to enhanced optical diagnosis of cancerous tissue. Moreover, quantitative assessment of the cytomorphological features visualized from MPM images showed significant differences in the nuclear-cytoplasmic ratios of pancreatic neoplasms compared with normal pancreas, as well as further distinguished pancreatic malignant tumors from benign tumors. These results indicate that the MPM could potentially serve as an optical tool for the diagnosis of pancreatic neoplasms in unstained histological sections.

  7. Combined multiphoton imaging and automated functional enucleation of porcine oocytes using femtosecond laser pulses

    NASA Astrophysics Data System (ADS)

    Kuetemeyer, Kai; Lucas-Hahn, Andrea; Petersen, Bjoern; Lemme, Erika; Hassel, Petra; Niemann, Heiner; Heisterkamp, Alexander

    2010-07-01

    Since the birth of ``Dolly'' as the first mammal cloned from a differentiated cell, somatic cell cloning has been successful in several mammalian species, albeit at low success rates. The highly invasive mechanical enucleation step of a cloning protocol requires sophisticated, expensive equipment and considerable micromanipulation skill. We present a novel noninvasive method for combined oocyte imaging and automated functional enucleation using femtosecond (fs) laser pulses. After three-dimensional imaging of Hoechst-labeled porcine oocytes by multiphoton microscopy, our self-developed software automatically identified the metaphase plate. Subsequent irradiation of the metaphase chromosomes with the very same laser at higher pulse energies in the low-density-plasma regime was used for metaphase plate ablation (functional enucleation). We show that fs laser-based functional enucleation of porcine oocytes completely inhibited the parthenogenetic development without affecting the oocyte morphology. In contrast, nonirradiated oocytes were able to develop parthenogenetically to the blastocyst stage without significant differences to controls. Our results indicate that fs laser systems have great potential for oocyte imaging and functional enucleation and may improve the efficiency of somatic cell cloning.

  8. Multiphoton Microscopy Applied for Real-Time Intravital Imaging of Bacterial Infections In Vivo

    PubMed Central

    Choong, Ferdinand X.; Sandoval, Ruben M.; Molitoris, Bruce A.; Richter-Dahlfors, Agneta

    2014-01-01

    To understand the underlying mechanisms of bacterial infections, researchers have for long addressed the molecular interactions occurring when the bacterium interacts with host target cells. In these studies, primarily based on in vitro systems, molecular details have been revealed along with increased knowledge regarding the general infection process. With the recent advancements in in vivo imaging techniques, we are now in a position to bridge a transition from classical minimalistic in vitro approaches to allow infections to be studied in its native complexity—the live organ. Techniques such as multiphoton microscopy (MPM) allow cellular-level visualization of the dynamic infection process in real time within the living host. Studies in which all interplaying factors, such as the influences of the immune, lymphatic, and vascular systems can be accounted for, are likely to provide new insights to our current understanding of the infection process. MPM imaging becomes extra powerful when combined with advanced surgical procedure, allowing studies of the illusive early hours of infection. In this chapter, our intention is to provide a general view on how to design and carry out intravital imaging of a bacterial infection. While exemplifying this using a spatiotemporally well-controlled uropathogenic Escherichia coli (UPEC) infection in rat kidneys, we hope to provide the reader with general considerations that can be adapted to other bacterial infections in organs other than the kidney. PMID:22341218

  9. Use of multiphoton tomography and fluorescence lifetime imaging to investigate skin pigmentation in vivo

    NASA Astrophysics Data System (ADS)

    Dancik, Yuri; Favre, Amandine; Loy, Chong Jin; Zvyagin, Andrei V.; Roberts, Michael S.

    2013-02-01

    There is a growing body of literature showing the usefulness of multiphoton tomography (MPT) and fluorescence lifetime imaging for in situ characterization of skin constituents and the ensuing development of noninvasive diagnostic tools against skin diseases. Melanin and pigmentation-associated skin cancers constitute some of the major applications. We show that MPT and fluorescence lifetime imaging can be used to measure changes in cutaneous melanin concentration and that these can be related to the visible skin color. Melanin in the skin of African, Indian, Caucasian, and Asian volunteers is detected on the basis of its emission wavelength and fluorescence lifetimes in solution and in a melanocyte-keratinocyte cell culture. Fluorescence intensity is used to characterize the melanin content and distribution as a function of skin type and depth into the skin (stratum granulosum and stratum basale). The measured fluorescence intensities in given skin types agree with melanin amounts reported by others using biopsies. Our results suggest that spatial distribution of melanin in skin can be studied using MPT and fluorescence lifetime imaging, but further studies are needed to ascertain that the method can resolve melanin amount in smaller depth intervals.

  10. Video-rate resonant scanning multiphoton microscopy: An emerging technique for intravital imaging of the tumor microenvironment.

    PubMed

    Kirkpatrick, Nathaniel D; Chung, Euiheon; Cook, Daniel C; Han, Xiaoxing; Gruionu, Gabriel; Liao, Shan; Munn, Lance L; Padera, Timothy P; Fukumura, Dai; Jain, Rakesh K

    2012-01-01

    The abnormal tumor microenvironment fuels tumor progression, metastasis, immune suppression, and treatment resistance. Over last several decades, developments in and applications of intravital microscopy have provided unprecedented insights into the dynamics of the tumor microenvironment. In particular, intravital multiphoton microscopy has revealed the abnormal structure and function of tumor-associated blood and lymphatic vessels, the role of aberrant tumor matrix in drug delivery, invasion and metastasis of tumor cells, the dynamics of immune cell trafficking to and within tumors, and gene expression in tumors. However, traditional multiphoton microscopy suffers from inherently slow imaging rates-only a few frames per second, thus unable to capture more rapid events such as blood flow, lymphatic flow, and cell movement within vessels. Here, we report the development and implementation of a video-rate multiphoton microscope (VR-MPLSM) based on resonant galvanometer mirror scanning that is capable of recording at 30 frames per second and acquiring intravital multispectral images. We show that the design of the system can be readily implemented and is adaptable to various experimental models. As examples, we demonstrate the utility of the system to directly measure flow within tumors, capture metastatic cancer cells moving within the brain vasculature and cells in lymphatic vessels, and image acute responses to changes in a vascular network. VR-MPLSM thus has the potential to further advance intravital imaging and provide new insight into the biology of the tumor microenvironment.

  11. Video-rate resonant scanning multiphoton microscopy: An emerging technique for intravital imaging of the tumor microenvironment.

    PubMed

    Kirkpatrick, Nathaniel D; Chung, Euiheon; Cook, Daniel C; Han, Xiaoxing; Gruionu, Gabriel; Liao, Shan; Munn, Lance L; Padera, Timothy P; Fukumura, Dai; Jain, Rakesh K

    2012-01-01

    The abnormal tumor microenvironment fuels tumor progression, metastasis, immune suppression, and treatment resistance. Over last several decades, developments in and applications of intravital microscopy have provided unprecedented insights into the dynamics of the tumor microenvironment. In particular, intravital multiphoton microscopy has revealed the abnormal structure and function of tumor-associated blood and lymphatic vessels, the role of aberrant tumor matrix in drug delivery, invasion and metastasis of tumor cells, the dynamics of immune cell trafficking to and within tumors, and gene expression in tumors. However, traditional multiphoton microscopy suffers from inherently slow imaging rates-only a few frames per second, thus unable to capture more rapid events such as blood flow, lymphatic flow, and cell movement within vessels. Here, we report the development and implementation of a video-rate multiphoton microscope (VR-MPLSM) based on resonant galvanometer mirror scanning that is capable of recording at 30 frames per second and acquiring intravital multispectral images. We show that the design of the system can be readily implemented and is adaptable to various experimental models. As examples, we demonstrate the utility of the system to directly measure flow within tumors, capture metastatic cancer cells moving within the brain vasculature and cells in lymphatic vessels, and image acute responses to changes in a vascular network. VR-MPLSM thus has the potential to further advance intravital imaging and provide new insight into the biology of the tumor microenvironment. PMID:24353926

  12. Imaging the morphological change of tissue structure during the early phase of esophageal tumor progression using multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Xu, Jian; Kang, Deyong; Xu, Meifang; Zhu, Xiaoqin; Zhuo, Shuangmu; Chen, Jianxin

    2012-12-01

    Esophageal cancer is a common malignancy with a very poor prognosis. Successful strategies for primary prevention and early detection are critically needed to control this disease. Multiphoton microscopy (MPM) is becoming a novel optical tool of choice for imaging tissue architecture and cellular morphology by two-photon excited fluorescence. In this study, we used MPM to image microstructure of human normal esophagus, carcinoma in situ (CIS), and early invasive carcinoma in order to establish the morphological features to differentiate these tissues. The diagnostic features such as the appearance of cancerous cells, the significant loss of stroma, the absence of the basement membrane were extracted to distinguish between normal and cancerous esophagus tissue. These results correlated well with the paired histological findings. With the advancement of clinically miniaturized MPM and the multi-photon probe, combining MPM with standard endoscopy will therefore allow us to make a real-time in vivo diagnosis of early esophageal cancer at the cellular level.

  13. [Thermal spectral property of prism in hyper spectral imager].

    PubMed

    Liang, Jiu-Sheng; Wu, Qing-Wen; Li, Ze-Xue; Chen, Li-Heng; Guo, Liang

    2010-06-01

    Prism is one of the most key parts in the hyper spectral imager (HSI). Consequently, to set thermal control target and make thermal control design, the thermal spectral property of prism in the HSI was studied. The working principle of the HSI and the definition of its thermal spectral property were introduced. The working environment of prism and its thermal effect were analyzed; also the study contents and technical route of the prism's thermal spectral property were discussed. The effects of different uniform temperature field on deflexion angle and angular dispersion of the prism in the HSI were deduced, and the changes in displacement of the spectra and the spectral bandwidth under different uniform temperature were obtained. For one instance, the thermal spectral property of the K9 prism and the fused silica prism were compared based on FEM and combined experiments, furthermore, its thermal control target was ascertained and a thermal spectral property test was carried out to validate the rationality of the thermal spectral property analysis. The results of analysis indicated that the changes in spectral bandwidth and spectrum resolution brought by thermal distortions can be ignored according to current fixing mode, and the displacement of the spectra is mainly determined by thermal coefficient of material refractive index; because of it's the lower thermal coefficient of material refractive index, the displacement of the spectra of the K9 prism is smaller under the same temperature changes; the material deflexion changes (dn/dlambda) of prism are not sensitive to the temperature, so the changes in spectral bandwidth caused by them are not obvious. And the results of test proved that the studied method of thermal spectral property is reasonable and essential, and the results are authentic and credible. So it can provide some guidance for setting thermal control target and optimizing thermal control design. PMID:20707180

  14. Investigation of signal-to-noise ratio in frequency-domain multiphoton fluorescence lifetime imaging microscopy.

    PubMed

    Zhang, Yide; Khan, Aamir A; Vigil, Genevieve D; Howard, Scott S

    2016-07-01

    Multiphoton microscopy (MPM) combined with fluorescence lifetime imaging microscopy (FLIM) has enabled three-dimensional quantitative molecular microscopy in vivo. The signal-to-noise ratio (SNR), and thus the imaging rate of MPM-FLIM, which is fundamentally limited by the shot noise and fluorescence saturation, has not been quantitatively studied yet. In this paper, we investigate the SNR performance of the frequency-domain (FD) MPM-FLIM with two figures of merit: the photon economy in the limit of shot noise, and the normalized SNR in the limit of saturation. The theoretical results and Monte Carlo simulations find that two-photon FD-FLIM requires 50% fewer photons to achieve the same SNR as conventional one-photon FLIM. We also analytically show that the MPM-FD-FLIM can exploit the DC and higher harmonic components generated by nonlinear optical mixing of the excitation light to improve SNR, reducing the required number of photons by an additional 50%. Finally, the effect of fluorophore saturation on the experimental SNR performance is discussed. PMID:27409702

  15. Imaging Mitochondrial Organization in Living Primate Oocytes and Embryos using Multiphoton Microscopy

    PubMed Central

    Squirrell, J.M.; Schramm, R.D.; Paprocki, A.M.; Wokosin, D.L.; Bavister, B.D.

    2016-01-01

    We employed multiphoton laser scanning microscopy (MPLSM) to image changes in mitochondrial distribution in living rhesus monkey embryos. This method of imaging does not impair development; thus, the same specimen can be visualized multiple times at various developmental stages. Not only does this increase the amount of information that can be gathered on a single specimen but it permits the correlation of early events with subsequent development in the same specimen. Here we demonstrate the utility of MPLSM for determining changes in mitochondrial organization at various developmental stages and show that rhesus zygotes possess a distinct accumulation of mitochondria between the pronuclei prior to syngamy. We present evidence that suggests that this pronuclear accumulation may be positively correlated with development to the blastocyst stage—in the same embryo—thereby illustrating how MPLSM can be used to correlate cellular dynamics of primate oocytes and early embryos with their developmental potential. Understanding the relationship between mitochondrial distribution and the subsequent development of mammalian embryos, particularly primates, will increase our ability to improve embryo culture technologies, including those used for human assisted reproduction. PMID:12807671

  16. Real-time imaging of the living kidney juxtaglomerular apparatus with multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Peti-Peterdi, Janos; Morishima, Shigeru; Bell, P. D.; Okada, Yasunobu

    2002-06-01

    The process of glomerular filtrate formation and regulation of renal hemodynamics, including the tubuloglomerular feedback (TGF) mechanism from the macula densa (MD) and renin release, involves the complex interaction of a number of different cell types of the juxtaglomerular apparatus (JGA). It has been difficult to study these cellular interactions in living preparations given the constraints of existing technologies. Recently, two photon confocal laser microscopy has been developed that offers a tremendous increase in optical resolution versus conventional confocal microscopy. Importantly it can optically section through an entire glomerulus (glomerular diameter approximately equals 100 micrometers ). Thus, it provides the ability to directly study structures and cellular components that lie deep within the glomerulus. This new technology was used in our studies. We now report high-resolution images of various glomerular and JGA cells using the membrane-marker TMA-DPH and the calcium fluorophore indo-1. Time-series images show how alterations in tubular fluid composition cause striking changes in single cell volume of the macula densa tubular epithelium in situ and how it also affects glomerular filtration through alterations in associated structures within the JGA. Multi-photon excitation fluorescence microscopy in combination with isolated perfused JGA offers a powerful new tool to investigate the structural and cellular components that regulate the process of glomerular filtrate formation and renal hemodynamics.

  17. Imaging Mitochondrial Organization in Living Primate Oocytes and Embryos using Multiphoton Microscopy

    NASA Astrophysics Data System (ADS)

    Squirrell, J. M.; Schramm, R. D.; Paprocki, A. M.; Wokosin, D. L.; Bavister, B. D.

    2003-06-01

    We employed multiphoton laser scanning microscopy (MPLSM) to image changes in mitochondrial distribution in living rhesus monkey embryos. This method of imaging does not impair development; thus, the same specimen can be visualized multiple times at various developmental stages. Not only does this increase the amount of information that can be gathered on a single specimen but it permits the correlation of early events with subsequent development in the same specimen. Here we demonstrate the utility of MPLSM for determining changes in mitochondrial organization at various developmental stages and show that rhesus zygotes possess a distinct accumulation of mitochondria between the pronuclei prior to syngamy. We present evidence that suggests that this pronuclear accumulation may be positively correlated with development to the blastocyst stage—in the same embryo—thereby illustrating how MPLSM can be used to correlate cellular dynamics of primate oocytes and early embryos with their developmental potential. Understanding the relationship between mitochondrial distribution and the subsequent development of mammalian embryos, particularly primates, will increase our ability to improve embryo culture technologies, including those used for human assisted reproduction.

  18. Spectral modulation interferometry for quantitative phase imaging

    PubMed Central

    Shang, Ruibo; Chen, Shichao; Li, Chengshuai; Zhu, Yizheng

    2015-01-01

    We propose a spectral-domain interferometric technique, termed spectral modulation interferometry (SMI), and present its application to high-sensitivity, high-speed, and speckle-free quantitative phase imaging. In SMI, one-dimensional complex field of an object is interferometrically modulated onto a broadband spectrum. Full-field phase and intensity images are obtained by scanning along the orthogonal direction. SMI integrates the high sensitivity of spectral-domain interferometry with the high speed of spectral modulation to quantify fast phase dynamics, and its dispersive and confocal nature eliminates laser speckles. The principle and implementation of SMI are discussed. Its performance is evaluated using static and dynamic objects. PMID:25780737

  19. Nonlinear spectral imaging of biological tissues

    NASA Astrophysics Data System (ADS)

    Palero, J. A.

    2007-07-01

    The work presented in this thesis demonstrates live high resolution 3D imaging of tissue in its native state and environment. The nonlinear interaction between focussed femtosecond light pulses and the biological tissue results in the emission of natural autofluorescence and second-harmonic signal. Because biological intrinsic emission is generally very weak and extends from the ultraviolet to the visible spectral range, a broad-spectral range and high sensitivity 3D spectral imaging system is developed. Imaging the spectral characteristics of the biological intrinsic emission reveals the structure and biochemistry of the cells and extra-cellular components. By using different methods in visualizing the spectral images, discrimination between different tissue structures is achieved without the use of any stain or fluorescent label. For instance, RGB real color spectral images of the intrinsic emission of mouse skin tissues show blue cells, green hair follicles, and purple collagen fibers. The color signature of each tissue component is directly related to its characteristic emission spectrum. The results of this study show that skin tissue nonlinear intrinsic emission is mainly due to the autofluorescence of reduced nicotinamide adenine dinucleotide (phosphate), flavins, keratin, melanin, phospholipids, elastin and collagen and nonlinear Raman scattering and second-harmonic generation in Type I collagen. In vivo time-lapse spectral imaging is implemented to study metabolic changes in epidermal cells in tissues. Optical scattering in tissues, a key factor in determining the maximum achievable imaging depth, is also investigated in this work.

  20. Iterative image reconstruction in spectral CT

    NASA Astrophysics Data System (ADS)

    Hernandez, Daniel; Michel, Eric; Kim, Hye S.; Kim, Jae G.; Han, Byung H.; Cho, Min H.; Lee, Soo Y.

    2012-03-01

    Scan time of spectral-CTs is much longer than conventional CTs due to limited number of x-ray photons detectable by photon-counting detectors. However, the spectral pixel information in spectral-CT has much richer information on physiological and pathological status of the tissues than the CT-number in conventional CT, which makes the spectral- CT one of the promising future imaging modalities. One simple way to reduce the scan time in spectral-CT imaging is to reduce the number of views in the acquisition of projection data. But, this may result in poorer SNR and strong streak artifacts which can severely compromise the image quality. In this work, spectral-CT projection data were obtained from a lab-built spectral-CT consisting of a single CdTe photon counting detector, a micro-focus x-ray tube and scan mechanics. For the image reconstruction, we used two iterative image reconstruction methods, the simultaneous iterative reconstruction technique (SIRT) and the total variation minimization based on conjugate gradient method (CG-TV), along with the filtered back-projection (FBP) to compare the image quality. From the imaging of the iodine containing phantoms, we have observed that SIRT and CG-TV are superior to the FBP method in terms of SNR and streak artifacts.

  1. Advanced multiphoton methods for in vitro and in vivo functional imaging of mouse retinal neurons (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Cohen, Noam; Schejter, Adi; Farah, Nairouz; Shoham, Shy

    2016-03-01

    Studying the responses of retinal ganglion cell (RGC) populations has major significance in vision research. Multiphoton imaging of optogenetic probes has recently become the leading approach for visualizing neural populations and has specific advantages for imaging retinal activity during visual stimulation, because it leads to reduced direct photoreceptor excitation. However, multiphoton retinal activity imaging is not straightforward: point-by-point scanning leads to repeated neural excitation while optical access through the rodent eye in vivo has proven highly challenging. Here, we present two enabling optical designs for multiphoton imaging of responses to visual stimuli in mouse retinas expressing calcium indicators. First, we present an imaging solution based on Scanning Line Temporal Focusing (SLITE) for rapidly imaging neuronal activity in vitro. In this design, we scan a temporally focused line rather than a point, increasing the scan speed and reducing the impact of repeated excitation, while maintaining high optical sectioning. Second, we present the first in vivo demonstration of two-photon imaging of RGC activity in the mouse retina. To obtain these cellular resolution recordings we integrated an illumination path into a correction-free imaging system designed using an optical model of the mouse eye. This system can image at multiple depths using an electronically tunable lens integrated into its optical path. The new optical designs presented here overcome a number of outstanding obstacles, allowing the study of rapid calcium- and potentially even voltage-indicator signals both in vitro and in vivo, thereby bringing us a step closer toward distributed monitoring of action potentials.

  2. Doppler imaging using spectrally-encoded endoscopy

    PubMed Central

    Yelin, Dvir; Bouma, B. E.; Rosowsky, J. J.; Tearney, G. J.

    2009-01-01

    The capability to image tissue motion such as blood flow through an endoscope could have many applications in medicine. Spectrally encoded endoscopy (SEE) is a recently introduced technique that utilizes a single optical fiber and miniature diffractive optics to obtain endoscopic images through small diameter probes. Using spectral-domain interferometry, SEE is furthermore capable of three-dimensional volume imaging at video rates. Here we show that by measuring relative spectral phases, this technology can additionally measure Doppler shifts. Doppler SEE is demonstrated in flowing Intralipid phantoms and vibrating middle ear ossicles. PMID:18795020

  3. New applications of Spectral Edge image fusion

    NASA Astrophysics Data System (ADS)

    Hayes, Alex E.; Montagna, Roberto; Finlayson, Graham D.

    2016-05-01

    In this paper, we present new applications of the Spectral Edge image fusion method. The Spectral Edge image fusion algorithm creates a result which combines details from any number of multispectral input images with natural color information from a visible spectrum image. Spectral Edge image fusion is a derivative-based technique, which creates an output fused image with gradients which are an ideal combination of those of the multispectral input images and the input visible color image. This produces both maximum detail and natural colors. We present two new applications of Spectral Edge image fusion. Firstly, we fuse RGB-NIR information from a sensor with a modified Bayer pattern, which captures visible and near-infrared image information on a single CCD. We also present an example of RGB-thermal image fusion, using a thermal camera attached to a smartphone, which captures both visible and low-resolution thermal images. These new results may be useful for computational photography and surveillance applications.

  4. Spectral OCT techniques in eye imaging

    NASA Astrophysics Data System (ADS)

    Kowalczyk, Andrzej; Wojtkowski, Maciej

    2002-02-01

    This contribution presents examples of images of eye in vitro obtained by spectral optical tomography (OCT). Particular interest was focused on obtaining clear images of the corneo-scleral angle and images of fundus which are both essential for diagnosing and planning of a treatment of glaucoma.

  5. Fibrillogenesis from nanosurfaces: multiphoton imaging and stereological analysis of collagen 3D self-assembly dynamics.

    PubMed

    Bancelin, Stéphane; Decencière, Etienne; Machairas, Vaïa; Albert, Claire; Coradin, Thibaud; Schanne-Klein, Marie-Claire; Aimé, Carole

    2014-09-21

    The assembly of proteins into fibrillar structures is an important process that concerns different biological contexts, including molecular medicine and functional biomaterials. Engineering of hybrid biomaterials can advantageously provide synergetic interactions of the biopolymers with an inorganic component to ensure specific supramolecular organization and dynamics. To this aim, we designed hybrid systems associating collagen and surface-functionalized silica particles and we built a new strategy to investigate fibrillogenesis processes in such multicomponents systems, working at the crossroads of chemistry, physics and mathematics. The self-assembly process was investigated by bimodal multiphoton imaging coupling second harmonic generation (SHG) and 2 photon excited fluorescence (2PEF). The in-depth spatial characterization of the system was further achieved using the three-dimensional analysis of the SHG/2PEF data via mathematical morphology processing. Quantitation of collagen distribution around particles offers strong evidence that the chemically induced confinement of the protein on the silica nanosurfaces has a key influence on the spatial extension of fibrillogenesis. This new approach is unique in the information it can provide on 3D dynamic hybrid systems and may be extended to other associations of fibrillar molecules with optically responsive nano-objects. PMID:25058449

  6. Controlled Au-Polymer Nanostructures for Multiphoton Imaging, Prodrug Delivery, and Chemo-Photothermal Therapy Platforms.

    PubMed

    Huang, Chih-Chia; Liu, Tzu-Ming

    2015-11-18

    We have successfully introduced a proton-induced controlled reaction of HAuCl4 and poly(styrene-alt-maleic acid) (PSMA) sodium salt to prepare triangular and multicore Au@polymer nanoparticles (NPs). The interparticle interactions in the core gave rise to an absorption band at the near-infrared wavelength. The near-infrared optical properties of the resulting Au-polymer nanostructures are highly stable in a physiological environment, which offered strong photo-to-thermal conversion by a moderate continuous-wave 808 nm laser and exhibited multiphoton fluorescence for imaging using a 1230 nm light excitation (femtosecond laser). Exposure of the carboxylate groups at the polymer shell made the surface structure of the Au multicore @polymer NPs directly conjugate Pt(II)-/Pt(IV)-based drugs, which possessed the elimination of the immediate toxicity over the short time and resulted in an anticancer effect after 3 days. A synergistic effect of the chemo-photothermal therapy showed a moderate hyperthermia assistance (<1 W/cm(2)) and better anticancer performance over time compared with the individual treatments. We demonstrated that such PSMA-based methodology not only enables a broad range of chemical material synthesis in the kinetic control to form Au nano-octahedrons and nanotriangles using Br(-)/I(-) ions additives but also could be extended to form Au/Fe3O4@polymer nanocomposites via proton-assisted PSMA self-assembly. PMID:26501876

  7. Multi-spectral compressive snapshot imaging using RGB image sensors.

    PubMed

    Rueda, Hoover; Lau, Daniel; Arce, Gonzalo R

    2015-05-01

    Compressive sensing is a powerful sensing and reconstruction framework for recovering high dimensional signals with only a handful of observations and for spectral imaging, compressive sensing offers a novel method of multispectral imaging. Specifically, the coded aperture snapshot spectral imager (CASSI) system has been demonstrated to produce multi-spectral data cubes color images from a single snapshot taken by a monochrome image sensor. In this paper, we expand the theoretical framework of CASSI to include the spectral sensitivity of the image sensor pixels to account for color and then investigate the impact on image quality using either a traditional color image sensor that spatially multiplexes red, green, and blue light filters or a novel Foveon image sensor which stacks red, green, and blue pixels on top of one another. PMID:25969307

  8. Spectral calibration of programmable imaging spectrometer

    NASA Astrophysics Data System (ADS)

    Du, Guojun; Liao, Zhibo; Jiao, Wenchun; Zong, Xiaoying; He, Xuhua; Wang, Haichao

    2015-10-01

    Programmable imager spectrometer can provide flexible data by changing the spectrum section number, central wavelength, spectral width and spatial resolution in orbit. Spectral calibration of imaging spectrometer plays an important role for acquiring accurate spectrum, two spectral calibration types are in essence: wavelength calibration and Full-width-half-maximum (FWHM). Base on the character of programmable imager spectrometer, designed a set of spectral calibration system. Wavelength calibration realized by utilizing the Monochromatic light of high precision monochromator, during the test, changed output parameters of monochromator according to the spectral bandwidth of imager spectrometer. The FWHM is constructed by a set of variable narrow spectrum lines that is output by tunable laser. Gaussian fitting algorithm is used to determine center wavelength and the FWHM of the characteristic spectrum line, Spectral pixels are calibrated by quadratic polynomial, standard spectroscopic lamp is used to verify wavelength calibration result accuracy. The calibration result indicates that FWHM is better than 2nm, the wavelength uncertainty is less than 0.6nm, meet the calibration requirements of programmable imaging spectrometer.

  9. Novel techniques with multiphoton microscopy: Deep-brain imaging with microprisms, neurometabolism of epilepsy, and counterfeit paper money detection

    NASA Astrophysics Data System (ADS)

    Chia, Thomas H.

    Multiphoton microscopy is a laser-scanning fluorescence imaging method with extraordinary potential. We describe three innovative multiphoton microscopy techniques across various disciplines. Traditional in vivo fluorescence microscopy of the mammalian brain has a limited penetration depth (<400 microm). We present a method of imaging 1 mm deep into mouse neocortex by using a glass microprism to relay the excitation and emission light. This technique enables simultaneous imaging of multiple cortical layers, including layer V, at an angle typical of slice preparations. At high-magnification imaging using an objective with 1-mm of coverglass correction, resolution was sufficient to resolve dendritic spines on layer V GFP neurons. Functional imaging of blood flow at various neocortical depths is also presented, allowing for quantification of red blood cell flux and velocity. Multiphoton fluorescence lifetime imaging (FLIM) of NADH reveals information on neurometabolism. NADH, an intrinsic fluorescent molecule and ubiquitous metabolic coenzyme, has a lifetime dependent on enzymatic binding. A novel NADH FLIM algorithm is presented that produces images showing spatially distinct NADH fluorescence lifetimes in mammalian brain slices. This program provides advantages over traditional FLIM processing of multi-component lifetime data. We applied this technique to a GFP-GFAP pilocarpine mouse model of temporal lobe epilepsy. Results indicated significant changes in the neurometabolism of astrocytes and neuropil in the cell and dendritic layers of the hippocampus when compared to control tissue. Data obtained with NADH FLIM were subsequently interpreted based on the abnormal activity reported in epileptic tissue. Genuine U.S. Federal Reserve Notes have a consistent, two-component intrinsic fluorescence lifetime. This allows for detection of counterfeit paper money because of its significant differences in fluorescence lifetime when compared to genuine paper money. We used

  10. Determining the Spectral Content of MOSES Images

    NASA Astrophysics Data System (ADS)

    Parker, Jacob; Kankelborg, Charles

    2016-05-01

    The MOSES (Multi-Order Solar EUV Spectrograph) sounding rocket was launched February 8th, 2006. The MOSES concave grating forms solar images in multiple spectral orders, in an effort to measure line profiles from a single exposure over a wide field of view. We present a preliminary identification of spectral content in MOSES images. The cross correlation of subtracted images provide evidence of spectral content besides the normal 304 Anstrom He II line. We place confidence on the peaks in correlation by cross correlating random data that is statistically representative of MOSES data. These significant peaks indicate a contribution to intensity from several coronal lines. These lines are individually weak, but if not taken into account, they would significantly increase the residuals when inverting MOSES images to obtain spectra.

  11. Spectral calibration for convex grating imaging spectrometer

    NASA Astrophysics Data System (ADS)

    Zhou, Jiankang; Chen, Xinhua; Ji, Yiqun; Chen, Yuheng; Shen, Weimin

    2013-12-01

    Spectral calibration of imaging spectrometer plays an important role for acquiring target accurate spectrum. There are two spectral calibration types in essence, the wavelength scanning and characteristic line sampling. Only the calibrated pixel is used for the wavelength scanning methods and he spectral response function (SRF) is constructed by the calibrated pixel itself. The different wavelength can be generated by the monochromator. The SRF is constructed by adjacent pixels of the calibrated one for the characteristic line sampling methods. And the pixels are illuminated by the narrow spectrum line and the center wavelength of the spectral line is exactly known. The calibration result comes from scanning method is precise, but it takes much time and data to deal with. The wavelength scanning method cannot be used in field or space environment. The characteristic line sampling method is simple, but the calibration precision is not easy to confirm. The standard spectroscopic lamp is used to calibrate our manufactured convex grating imaging spectrometer which has Offner concentric structure and can supply high resolution and uniform spectral signal. Gaussian fitting algorithm is used to determine the center position and the Full-Width-Half-Maximum(FWHM)of the characteristic spectrum line. The central wavelengths and FWHMs of spectral pixels are calibrated by cubic polynomial fitting. By setting a fitting error thresh hold and abandoning the maximum deviation point, an optimization calculation is achieved. The integrated calibration experiment equipment for spectral calibration is developed to enhance calibration efficiency. The spectral calibration result comes from spectral lamp method are verified by monochromator wavelength scanning calibration technique. The result shows that spectral calibration uncertainty of FWHM and center wavelength are both less than 0.08nm, or 5.2% of spectral FWHM.

  12. Information-efficient spectral imaging sensor

    DOEpatents

    Sweatt, William C.; Gentry, Stephen M.; Boye, Clinton A.; Grotbeck, Carter L.; Stallard, Brian R.; Descour, Michael R.

    2003-01-01

    A programmable optical filter for use in multispectral and hyperspectral imaging. The filter splits the light collected by an optical telescope into two channels for each of the pixels in a row in a scanned image, one channel to handle the positive elements of a spectral basis filter and one for the negative elements of the spectral basis filter. Each channel for each pixel disperses its light into n spectral bins, with the light in each bin being attenuated in accordance with the value of the associated positive or negative element of the spectral basis vector. The spectral basis vector is constructed so that its positive elements emphasize the presence of a target and its negative elements emphasize the presence of the constituents of the background of the imaged scene. The attenuated light in the channels is re-imaged onto separate detectors for each pixel and then the signals from the detectors are combined to give an indication of the presence or not of the target in each pixel of the scanned scene. This system provides for a very efficient optical determination of the presence of the target, as opposed to the very data intensive data manipulations that are required in conventional hyperspectral imaging systems.

  13. Multi Spectral Fluorescence Imager (MSFI)

    NASA Technical Reports Server (NTRS)

    Caron, Allison

    2016-01-01

    Genetic transformation with in vivo reporter genes for fluorescent proteins can be performed on a variety of organisms to address fundamental biological questions. Model organisms that may utilize an ISS imager include unicellular organisms (Saccharomyces cerevisiae), plants (Arabidopsis thaliana), and invertebrates (Caenorhabditis elegans). The multispectral fluorescence imager (MSFI) will have the capability to accommodate 10 cm x 10 cm Petri plates, various sized multi-well culture plates, and other custom culture containers. Features will include programmable temperature and light cycles, ethylene scrubbing (less than 25 ppb), CO2 control (between 400 ppm and ISS-ambient levels in units of 100 ppm) and sufficient airflow to prevent condensation that would interfere with imaging.

  14. Multiphoton tomography of intratissue tattoo nanoparticles

    NASA Astrophysics Data System (ADS)

    König, Karsten

    2012-02-01

    Most of today's intratissue tattoo pigments are unknown nanoparticles. So far, there was no real control of their use due to the absence of regulations. Some of the tattoo pigments contain carcinogenic amines e.g. azo pigment Red 22. Nowadays, the European Union starts to control the administration of tattoo pigments. There is an interest to obtain information on the intratissue distribution, their interaction with living cells and the extracellular matrix, and the mechanisms behind laser tattoo removal. Multiphoton tomographs are novel biosafety and imaging tools that can provide such information non-invasively and without further labeling. When using the spectral FLIM module, spatially-resolved emission spectra, excitation spectra, and fluorescence lifetimes can pr provided. Multiphoton tomographs are used by all major cosmetic comapanies to test the biosafety of sunscreen nanoparticles.

  15. MULTIPHOTON PROCESSES

    SciTech Connect

    2002-07-05

    The Gordon Research Conference (GRC) on MULTIPHOTON PROCESSES was held at Tilton School, Tilton, NH. Emphasis was placed on current unpublished research and discussion of the future target areas in this field.

  16. Spectrum slicer for snapshot spectral imaging

    NASA Astrophysics Data System (ADS)

    Tamamitsu, Miu; Kitagawa, Yutaro; Nakagawa, Keiichi; Horisaki, Ryoichi; Oishi, Yu; Morita, Shin-ya; Yamagata, Yutaka; Motohara, Kentaro; Goda, Keisuke

    2015-12-01

    We propose and demonstrate an optical component that overcomes critical limitations in our previously demonstrated high-speed multispectral videography-a method in which an array of periscopes placed in a prism-based spectral shaper is used to achieve snapshot multispectral imaging with the frame rate only limited by that of an image-recording sensor. The demonstrated optical component consists of a slicing mirror incorporated into a 4f-relaying lens system that we refer to as a spectrum slicer (SS). With its simple design, we can easily increase the number of spectral channels without adding fabrication complexity while preserving the capability of high-speed multispectral videography. We present a theoretical framework for the SS and its experimental utility to spectral imaging by showing real-time monitoring of a dynamic colorful event through five different visible windows.

  17. Label-free imaging of goblet cells as a marker for differentiating colonic polyps by multiphoton microscopy Label-free imaging of goblet cells

    NASA Astrophysics Data System (ADS)

    Zhuo, S. M.; Wu, G. Z.; Chen, J. X.; Zhu, X. Q.; Xie, S. S.

    2012-06-01

    Discrimination of adenomas from hyperplastic polyps can reduce the risk of unnecessary complications and healthcare cost. However, it is challenging during colonoscopy screening, and histological analysis remains the ``gold standard'' for the final diagnosis. Here, we describe a label-free imaging method, multiphoton microscopy (MPM), to the discrimination between adenomas and hyperplastic polyps. We find that multiphoton imaging provides cellular and subcellular details to the identification of adenomas from hyperplastic polyps. In particular, there is significant difference in the population density of goblet cells among normal colon, hyperplastic polyp, and adenoma, providing substantial potential to become a quantitative intrinsic marker for in vivo clinical diagnosis of early colonic lesions. To our knowledge, this is the first demonstration of the potential of MPM for differentiation of colonic polyps.

  18. Calibration and characterization of spectral imaging systems

    NASA Astrophysics Data System (ADS)

    Polder, Gerrit; van der Heijden, Gerie W.

    2001-09-01

    Spectral image sensors provide images with a large umber of contiguous spectral channels per pixel. This paper describes the calibration of spectrograph based spectral imaging systems. The relation between pixel position and measured wavelength was determined using three different wavelength calibration sources. Results indicate that for spectral calibration a source with very small peaks,such as a HgAr source, is preferred to arrow band filters. A second order polynomial model gives a better fit than a linear model for the pixel to wavelength mapping. The signal to noise ratio (SNR)is determined per wavelength. In the blue part of the spectrum,the SNR was lower than in the green and red part.This is due to a decreased quantum efficiency of the CCD,a smaller transmission coefficient of the spectrograph,as well as poor performance of the illuminant. Increasing the amount of blue light,using additional Fluorescent tube with special coating increased the SNR considerably. Furthermore, the spatial and spectral resolution of the system are determined.These can be used to choose appropriate binning factors to decrease the image size without losing information.

  19. Dynamical Spectral Unmixing of Multitemporal Hyperspectral Images

    NASA Astrophysics Data System (ADS)

    Henrot, Simon; Chanussot, Jocelyn; Jutten, Christian

    2016-07-01

    In this paper, we consider the problem of unmixing a time series of hyperspectral images. We propose a dynamical model based on linear mixing processes at each time instant. The spectral signatures and fractional abundances of the pure materials in the scene are seen as latent variables, and assumed to follow a general dynamical structure. Based on a simplified version of this model, we derive an efficient spectral unmixing algorithm to estimate the latent variables by performing alternating minimizations. The performance of the proposed approach is demonstrated on synthetic and real multitemporal hyperspectral images.

  20. SPECTRAL SMILE CORRECTION IN CRISM HYPERSPECTRAL IMAGES

    NASA Astrophysics Data System (ADS)

    Ceamanos, X.; Doute, S.

    2009-12-01

    The Compact Reconnaissance Imaging Spectrometer for Mars (CRISM) is affected by a common artifact in "push-broom" sensors, the so-called "spectral smile". As a consequence, both central wavelength and spectral width of the spectral response vary along the across-track dimension, thus giving rise to a shifting and smoothing of spectra (see Fig. 1 (left)). In fact, both effects are greater for spectra on the edges, while they are minimum for data acquired by central detectors, the so-called "sweet spot". The prior artifacts become particularly critical for Martian observations which contain steep spectra such as CO2 ice-rich polar images. Fig. 1 (right) shows the horizontal brightness gradient which appears in every band corresponding to a steep portion of spectra. The correction of CRISM spectral smile is addressed using a two-step method which aims at modifying data sensibly in order to mimic the optimal CRISM response. First, all spectra, which are previously interpolated by cubic splines, are resampled to the "sweet spot" wavelengths in order to overcome the spectra shift. Secondly, the non-uniform spectral width is overcome by mimicking an increase of spectral resolution thanks to a spectral sharpening. In order to minimize noise, only bands particularly suffering from smile are selected. First, bands corresponding to the outliers of the Minimum Noise Transformation (MNF) eigenvector, which corresponds to the MNF band related to smile (MNF-smile), are selected. Then, a spectral neighborhood Θi, which takes into account the local spectral convexity or concavity, is defined for every selected band in order to maximize spectral shape preservation. The proposed sharpening technique takes into account both the instrument parameters and the observed spectra. First, every reflectance value belonging to a Θi is reevaluated by a sharpening which depends on a ratio of the spectral width of the current detector and the "sweet spot" one. Then, the optimal degree of

  1. Remote Sensing of Landscapes with Spectral Images

    NASA Astrophysics Data System (ADS)

    Adams, John B.; Gillespie, Alan R.

    2006-05-01

    Remote Sensing of Landscapes with Spectral Images describes how to process and interpret spectral images using physical models to bridge the gap between the engineering and theoretical sides of remote-sensing and the world that we encounter when we venture outdoors. The emphasis is on the practical use of images rather than on theory and mathematical derivations. Examples are drawn from a variety of landscapes and interpretations are tested against the reality seen on the ground. The reader is led through analysis of real images (using figures and explanations); the examples are chosen to illustrate important aspects of the analytic framework. This textbook will form a valuable reference for graduate students and professionals in a variety of disciplines including ecology, forestry, geology, geography, urban planning, archeology and civil engineering. It is supplemented by a web-site hosting digital color versions of figures in the book as well as ancillary images (www.cambridge.org/9780521662214). Presents a coherent view of practical remote sensing, leading from imaging and field work to the generation of useful thematic maps Explains how to apply physical models to help interpret spectral images Supplemented by a website hosting digital colour versions of figures in the book, as well as additional colour figures

  2. Label-free multi-photon imaging using a compact femtosecond fiber laser mode-locked by carbon nanotube saturable absorber

    PubMed Central

    Kieu, K.; Mehravar, S.; Gowda, R.; Norwood, R. A.; Peyghambarian, N.

    2013-01-01

    We demonstrate label-free multi-photon imaging of biological samples using a compact Er3+-doped femtosecond fiber laser mode-locked by a single-walled carbon nanotube (CNT). These compact and low cost lasers have been developed by various groups but they have not been exploited for multiphoton microscopy. Here, it is shown that various multiphoton imaging modalities (e.g. second harmonic generation (SHG), third harmonic generation (THG), two-photon excitation fluorescence (TPEF), and three-photon excitation fluorescence (3PEF)) can be effectively performed on various biological samples using a compact handheld CNT mode-locked femtosecond fiber laser operating in the telecommunication window near 1560nm. We also show for the first time that chlorophyll fluorescence in plant leaves and diatoms can be observed using 1560nm laser excitation via three-photon absorption. PMID:24156074

  3. Label-free multi-photon imaging using a compact femtosecond fiber laser mode-locked by carbon nanotube saturable absorber.

    PubMed

    Kieu, K; Mehravar, S; Gowda, R; Norwood, R A; Peyghambarian, N

    2013-01-01

    We demonstrate label-free multi-photon imaging of biological samples using a compact Er(3+)-doped femtosecond fiber laser mode-locked by a single-walled carbon nanotube (CNT). These compact and low cost lasers have been developed by various groups but they have not been exploited for multiphoton microscopy. Here, it is shown that various multiphoton imaging modalities (e.g. second harmonic generation (SHG), third harmonic generation (THG), two-photon excitation fluorescence (TPEF), and three-photon excitation fluorescence (3PEF)) can be effectively performed on various biological samples using a compact handheld CNT mode-locked femtosecond fiber laser operating in the telecommunication window near 1560nm. We also show for the first time that chlorophyll fluorescence in plant leaves and diatoms can be observed using 1560nm laser excitation via three-photon absorption.

  4. Miniaturized spectral imager for Aalto-1 nanosatellite

    NASA Astrophysics Data System (ADS)

    Mannila, Rami; Näsilä, Antti; Praks, Jaan; Saari, Heikki; Antila, Jarkko

    2011-11-01

    The Aalto-1 is a 3U-cubesat project coordinated by Aalto University. The satellite, Aalto-1, will be mainly built by students as project assignments and thesis works. VTT Technical Research Centre of Finland will develop the main Earth observation payload, a miniaturized spectral imager, for the satellite. It is a novel highly miniaturized tunable filter type spectral imager. Mass of the spectral imager will be less than 400 grams, and dimensions will be approximately 80 mm x 80 mm x 45 mm. The spectral imager is based on a tunable Fabry-Pérot interferometer (FPI) accompanied by an RGB CMOS image sensor. The FPI consists of two highly reflective surfaces separated by a tunable air gap and it is based either on a microelectromechanical (MEMS) or piezo-actuated structure. The MEMS FPI is a monolithic device, i.e. it is made entirely on one substrate in a batch process, without assembling separate pieces together. The gap is adjusted by moving the upper mirror with electrostatic force. Benefits of the MEMS FPI are low mass and small size. However, large aperture (2-10 mm) MEMS FPIs are currently under development, thus it is not yet known if their performance is adequate. The piezo-actuated FPI uses three piezo-actuators and is controlled in a closed capacitive feedback loop. The drawback of the piezo-actuated FPI is its higher mass. However, it has a large aperture which enables a shorter exposure times. Selection of the FPI type will be done after thorough evaluation. Depending on the selected FPI type, the spectral resolution of the imager will be 5 - 10 nm at full width at half maximum and it will operate in the visible and/or near infrared range.

  5. Multi-spectral imaging of oxygen saturation

    NASA Astrophysics Data System (ADS)

    Savelieva, Tatiana A.; Stratonnikov, Aleksander A.; Loschenov, Victor B.

    2008-06-01

    The system of multi-spectral imaging of oxygen saturation is an instrument that can record both spectral and spatial information about a sample. In this project, the spectral imaging technique is used for monitoring of oxygen saturation of hemoglobin in human tissues. This system can be used for monitoring spatial distribution of oxygen saturation in photodynamic therapy, surgery or sports medicine. Diffuse reflectance spectroscopy in the visible range is an effective and extensively used technique for the non-invasive study and characterization of various biological tissues. In this article, a short review of modeling techniques being currently in use for diffuse reflection from semi-infinite turbid media is presented. A simple and practical model for use with a real-time imaging system is proposed. This model is based on linear approximation of the dependence of the diffuse reflectance coefficient on relation between absorbance and reduced scattering coefficient. This dependence was obtained with the Monte Carlo simulation of photon propagation in turbid media. Spectra of the oxygenated and deoxygenated forms of hemoglobin differ mostly in the red area (520 - 600 nm) and have several characteristic points there. Thus four band-pass filters were used for multi-spectral imaging. After having measured the reflectance, the data obtained are used for fitting the concentration of oxygenated and free hemoglobin, and hemoglobin oxygen saturation.

  6. Integrated spectrometer design with application to multiphoton microscopy.

    PubMed

    Chandler, Eric V; Durfee, Charles G; Squier, Jeffrey A

    2011-01-01

    We present a prism-based spectrometer integrated into a multifocal, multiphoton microscope. The multifocal configuration facilitates interrogation of samples under different excitation conditions. Notably, the image plane of the microscope and the image plane of the spectrometer are coincident eliminating the need for an intermediate image plane containing an entrance slit. An EM-CCD detector provides sufficient gain for spectral interrogation of single-emitters. We employ this spectrometer to observe spectral shifts in the two-photon excitation fluorescence emission of single CdSe nanodots as a function of excitation polarization. PMID:21263548

  7. High-fidelity spatially resolved multiphoton counting for quantum imaging applications.

    PubMed

    Chrapkiewicz, Radosław; Wasilewski, Wojciech; Banaszek, Konrad

    2014-09-01

    We present a method for spatially resolved multiphoton counting based on an intensified camera with the retrieval of multimode photon statistics fully accounting for nonlinearities in the detection process. The scheme relies on one-time quantum tomographic calibration of the detector. Faithful, high-fidelity reconstruction of single- and two-mode statistics of multiphoton states is demonstrated for coherent states and their statistical mixtures. The results consistently exhibit classical values of the Mandel parameter and the noise reduction factor in contrast to raw statistics of camera photo-events. Detector operation is reliable for illumination levels up to the average of one detected photon per an event area-substantially higher than in previous approaches to characterize quantum statistical properties of light with spatial resolution. PMID:25166081

  8. Analysis for simplified optics coma effection on spectral image inversion of coded aperture spectral imager

    NASA Astrophysics Data System (ADS)

    Liu, Yangyang; Lv, Qunbo; Li, Weiyan; Xiangli, Bin

    2015-09-01

    As a novel spectrum imaging technology was developed recent years, push-broom coded aperture spectral imaging (PCASI) has the advantages of high throughput, high SNR, high stability etc. This coded aperture spectral imaging utilizes fixed code templates and push-broom mode, which can realize the high-precision reconstruction of spatial and spectral information. But during optical lens designing, manufacturing and debugging, it is inevitably exist some minor coma errors. Even minor coma errors can reduce image quality. In this paper, we simulated the system optical coma error's influence to the quality of reconstructed image, analyzed the variant of the coded aperture in different optical coma effect, then proposed an accurate curve of image quality and optical coma quality in 255×255 size code template, which provide important references for design and development of push-broom coded aperture spectrometer.

  9. New procedure for capturing spectral images of human portraiture

    NASA Astrophysics Data System (ADS)

    Sun, Qun; Fairchild, Mark D.

    2002-06-01

    This paper describes a new procedure of capturing spectral images of human portraiture. The designed imaging system was calibrated directly based on real human subjects and has the capability to provide accurate spectral images of human faces, including facial skin as well as the lips, eyes, and hair, from various ethnic races. The facial spectral reflectances obtained were analyzed by principal components analysis (PCA) method. Based on the results of PCA, spectral images using both three and six wide-band spectral sampling were estimated. The reconstructed spectral images for display based on an sRGB display model are evaluated. The results have proved that this new spectral imaging procedure is successful. The results also show that three basis functions are accurate enough to estimate the spectral reflectance of human faces. The derived spectral images can be applied to color-imaging system design and analysis.

  10. WAVELENGTH AND ALIGNMENT TESTS FOR CONFOCAL SPECTRAL IMAGING SYSTEMS

    EPA Science Inventory

    Confocal spectral imaging (CSI) microscope systems now on the market delineate multiple fluorescent proteins, labels, or dyes within biological specimens by performing spectral characterizations. However, we find that some CSI present inconsistent spectral profiles of reference s...

  11. CALIBRATION AND VALIDATION OF CONFOCAL SPECTRAL IMAGING SYSTEMS

    EPA Science Inventory

    Confocal spectral imaging (CSI) microscope systems now on the market can perform spectral characterization of biological specimens containing fluorescent proteins, labels or dyes. Some CSI have been found to present inconsistent spectral characterizations within a particular syst...

  12. From morphology to biochemical state – intravital multiphoton fluorescence lifetime imaging of inflamed human skin

    NASA Astrophysics Data System (ADS)

    Huck, Volker; Gorzelanny, Christian; Thomas, Kai; Getova, Valentina; Niemeyer, Verena; Zens, Katharina; Unnerstall, Tim R.; Feger, Julia S.; Fallah, Mohammad A.; Metze, Dieter; Ständer, Sonja; Luger, Thomas A.; Koenig, Karsten; Mess, Christian; Schneider, Stefan W.

    2016-03-01

    The application of multiphoton microscopy in the field of biomedical research and advanced diagnostics promises unique insights into the pathophysiology of inflammatory skin diseases. In the present study, we combined multiphoton-based intravital tomography (MPT) and fluorescence lifetime imaging (MPT-FLIM) within the scope of a clinical trial of atopic dermatitis with the aim of providing personalised data on the aetiopathology of inflammation in a non-invasive manner at patients’ bedsides. These ‘optical biopsies’ generated via MPT were morphologically analysed and aligned with classical skin histology. Because of its subcellular resolution, MPT provided evidence of a redistribution of mitochondria in keratinocytes, indicating an altered cellular metabolism. Two independent morphometric algorithms reliably showed an even distribution in healthy skin and a perinuclear accumulation in inflamed skin. Moreover, using MPT-FLIM, detection of the onset and progression of inflammatory processes could be achieved. In conclusion, the change in the distribution of mitochondria upon inflammation and the verification of an altered cellular metabolism facilitate a better understanding of inflammatory skin diseases and may permit early diagnosis and therapy.

  13. From morphology to biochemical state – intravital multiphoton fluorescence lifetime imaging of inflamed human skin

    PubMed Central

    Huck, Volker; Gorzelanny, Christian; Thomas, Kai; Getova, Valentina; Niemeyer, Verena; Zens, Katharina; Unnerstall, Tim R.; Feger, Julia S.; Fallah, Mohammad A.; Metze, Dieter; Ständer, Sonja; Luger, Thomas A.; Koenig, Karsten; Mess, Christian; Schneider, Stefan W.

    2016-01-01

    The application of multiphoton microscopy in the field of biomedical research and advanced diagnostics promises unique insights into the pathophysiology of inflammatory skin diseases. In the present study, we combined multiphoton-based intravital tomography (MPT) and fluorescence lifetime imaging (MPT-FLIM) within the scope of a clinical trial of atopic dermatitis with the aim of providing personalised data on the aetiopathology of inflammation in a non-invasive manner at patients’ bedsides. These ‘optical biopsies’ generated via MPT were morphologically analysed and aligned with classical skin histology. Because of its subcellular resolution, MPT provided evidence of a redistribution of mitochondria in keratinocytes, indicating an altered cellular metabolism. Two independent morphometric algorithms reliably showed an even distribution in healthy skin and a perinuclear accumulation in inflamed skin. Moreover, using MPT-FLIM, detection of the onset and progression of inflammatory processes could be achieved. In conclusion, the change in the distribution of mitochondria upon inflammation and the verification of an altered cellular metabolism facilitate a better understanding of inflammatory skin diseases and may permit early diagnosis and therapy. PMID:27004454

  14. Multiphoton microscopic imaging of histological sections without hematoxylin and eosin staining differentiates carcinoma in situ lesion from normal oesophagus

    NASA Astrophysics Data System (ADS)

    Chen, Jianxin; Xu, Jian; Kang, Deyong; Xu, Meifang; Zhuo, Shuangmu; Zhu, Xiaoqin; Jiang, Xingshan

    2013-10-01

    Multiphoton microscopy (MPM) has become a powerful, important tool for tissues imaging at the molecular level. In this paper, this technique was extended to histological investigations, differentiating carcinoma in situ (CIS) lesion from normal oesophagus by imaging histological sections without hematoxylin and eosin (H&E) staining. The results show that the histology procedures of dehydration, paraffin embedding, and de-paraffinizing highlighted two photon excited fluorescence of cytoplasm and nucleolus of epithelial cell and collagen in stroma. MPM has the ability to identify the characteristics of CIS lesion including changes of squamous cells and full epithelium, identification of basement membrane, especially prominent nucleolus. The studies described here show that MPM has the potential for future retrospective studies of tumor staging by employing on histological section specimens without H&E staining.

  15. The spectral imaging facility: Setup characterization

    SciTech Connect

    De Angelis, Simone De Sanctis, Maria Cristina; Manzari, Paola Olga; Ammannito, Eleonora; Di Iorio, Tatiana; Liberati, Fabrizio; Tarchi, Fabio; Dami, Michele; Olivieri, Monica; Pompei, Carlo; Mugnuolo, Raffaele

    2015-09-15

    The SPectral IMager (SPIM) facility is a laboratory visible infrared spectrometer developed to support space borne observations of rocky bodies of the solar system. Currently, this laboratory setup is used to support the DAWN mission, which is in its journey towards the asteroid 1-Ceres, and to support the 2018 Exo-Mars mission in the spectral investigation of the Martian subsurface. The main part of this setup is an imaging spectrometer that is a spare of the DAWN visible infrared spectrometer. The spectrometer has been assembled and calibrated at Selex ES and then installed in the facility developed at the INAF-IAPS laboratory in Rome. The goal of SPIM is to collect data to build spectral libraries for the interpretation of the space borne and in situ hyperspectral measurements of planetary materials. Given its very high spatial resolution combined with the imaging capability, this instrument can also help in the detailed study of minerals and rocks. In this paper, the instrument setup is first described, and then a series of test measurements, aimed to the characterization of the main subsystems, are reported. In particular, laboratory tests have been performed concerning (i) the radiation sources, (ii) the reference targets, and (iii) linearity of detector response; the instrumental imaging artifacts have also been investigated.

  16. The spectral imaging facility: Setup characterization

    NASA Astrophysics Data System (ADS)

    De Angelis, Simone; Ammannito, Eleonora; Di Iorio, Tatiana; De Sanctis, Maria Cristina; Manzari, Paola Olga; Liberati, Fabrizio; Tarchi, Fabio; Dami, Michele; Olivieri, Monica; Pompei, Carlo; Mugnuolo, Raffaele

    2015-09-01

    The SPectral IMager (SPIM) facility is a laboratory visible infrared spectrometer developed to support space borne observations of rocky bodies of the solar system. Currently, this laboratory setup is used to support the DAWN mission, which is in its journey towards the asteroid 1-Ceres, and to support the 2018 Exo-Mars mission in the spectral investigation of the Martian subsurface. The main part of this setup is an imaging spectrometer that is a spare of the DAWN visible infrared spectrometer. The spectrometer has been assembled and calibrated at Selex ES and then installed in the facility developed at the INAF-IAPS laboratory in Rome. The goal of SPIM is to collect data to build spectral libraries for the interpretation of the space borne and in situ hyperspectral measurements of planetary materials. Given its very high spatial resolution combined with the imaging capability, this instrument can also help in the detailed study of minerals and rocks. In this paper, the instrument setup is first described, and then a series of test measurements, aimed to the characterization of the main subsystems, are reported. In particular, laboratory tests have been performed concerning (i) the radiation sources, (ii) the reference targets, and (iii) linearity of detector response; the instrumental imaging artifacts have also been investigated.

  17. The spectral imaging facility: Setup characterization.

    PubMed

    De Angelis, Simone; Ammannito, Eleonora; Di Iorio, Tatiana; De Sanctis, Maria Cristina; Manzari, Paola Olga; Liberati, Fabrizio; Tarchi, Fabio; Dami, Michele; Olivieri, Monica; Pompei, Carlo; Mugnuolo, Raffaele

    2015-09-01

    The SPectral IMager (SPIM) facility is a laboratory visible infrared spectrometer developed to support space borne observations of rocky bodies of the solar system. Currently, this laboratory setup is used to support the DAWN mission, which is in its journey towards the asteroid 1-Ceres, and to support the 2018 Exo-Mars mission in the spectral investigation of the Martian subsurface. The main part of this setup is an imaging spectrometer that is a spare of the DAWN visible infrared spectrometer. The spectrometer has been assembled and calibrated at Selex ES and then installed in the facility developed at the INAF-IAPS laboratory in Rome. The goal of SPIM is to collect data to build spectral libraries for the interpretation of the space borne and in situ hyperspectral measurements of planetary materials. Given its very high spatial resolution combined with the imaging capability, this instrument can also help in the detailed study of minerals and rocks. In this paper, the instrument setup is first described, and then a series of test measurements, aimed to the characterization of the main subsystems, are reported. In particular, laboratory tests have been performed concerning (i) the radiation sources, (ii) the reference targets, and (iii) linearity of detector response; the instrumental imaging artifacts have also been investigated. PMID:26429423

  18. Automated eXpert Spectral Image Analysis

    2003-11-25

    AXSIA performs automated factor analysis of hyperspectral images. In such images, a complete spectrum is collected an each point in a 1-, 2- or 3- dimensional spatial array. One of the remaining obstacles to adopting these techniques for routine use is the difficulty of reducing the vast quantities of raw spectral data to meaningful information. Multivariate factor analysis techniques have proven effective for extracting the essential information from high dimensional data sets into a limtedmore » number of factors that describe the spectral characteristics and spatial distributions of the pure components comprising the sample. AXSIA provides tools to estimate different types of factor models including Singular Value Decomposition (SVD), Principal Component Analysis (PCA), PCA with factor rotation, and Alternating Least Squares-based Multivariate Curve Resolution (MCR-ALS). As part of the analysis process, AXSIA can automatically estimate the number of pure components that comprise the data and can scale the data to account for Poisson noise. The data analysis methods are fundamentally based on eigenanalysis of the data crossproduct matrix coupled with orthogonal eigenvector rotation and constrained alternating least squares refinement. A novel method for automatically determining the number of significant components, which is based on the eigenvalues of the crossproduct matrix, has also been devised and implemented. The data can be compressed spectrally via PCA and spatially through wavelet transforms, and algorithms have been developed that perform factor analysis in the transform domain while retaining full spatial and spectral resolution in the final result. These latter innovations enable the analysis of larger-than core-memory spectrum-images. AXSIA was designed to perform automated chemical phase analysis of spectrum-images acquired by a variety of chemical imaging techniques. Successful applications include Energy Dispersive X-ray Spectroscopy, X

  19. Detection of latent fingerprints by ultraviolet spectral imaging

    NASA Astrophysics Data System (ADS)

    Huang, Wei; Xu, Xiaojing; Wang, Guiqiang

    2013-12-01

    Spectral imaging technology research is becoming more popular in the field of forensic science. Ultraviolet spectral imaging technology is an especial part of the full spectrum of imaging technology. This paper finished the experiment contents of the ultraviolet spectrum imaging method and image acquisition system based on ultraviolet spectral imaging technology. Ultraviolet spectral imaging experiments explores a wide variety of ultraviolet reflectance spectra of the object material curve and its ultraviolet spectrum of imaging modalities, can not only gives a reference for choosing ultraviolet wavelength to show the object surface potential traces of substances, but also gives important data for the ultraviolet spectrum of imaging technology development.

  20. Multicolor multiphoton microscopy based on a nanosecond supercontinuum laser source.

    PubMed

    Lefort, Claire; O'Connor, Rodney P; Blanquet, Véronique; Magnol, Laetitia; Kano, Hideaki; Tombelaine, Vincent; Lévêque, Philippe; Couderc, Vincent; Leproux, Philippe

    2016-07-01

    Multicolor multiphoton microscopy is experimentally demonstrated for the first time on a spectral bandwidth of excitation of 300 nm (full width half maximum) thanks to the implementation a nanosecond supercontinuum (SC) source compact and simple with a low repetition rate. The interest of such a wide spectral bandwidth, never demonstrated until now, is highlighted in vivo: images of glioma tumor cells stably expressing eGFP grafted on the brain of a mouse and its blood vessels network labelled with Texas Red(®) are obtained. These two fluorophores have a spectral bandwidth covering the whole 300 nm available. In parallel, a similar image quality is obtained on a sample of mouse muscle in vitro when excited with this nanosecond SC source or with a classical high rate, femtosecond and quasi monochromatic laser. This opens the way for (i) a simple and very complete biological characterization never performed to date with multiphoton processes, (ii) multiple means of contrast in nonlinear imaging allowed by the use of numerous fluorophores and (iii) other multiphoton processes like three-photon ones. PMID:26872004

  1. [Microscopic infrared spectral imaging of oily core].

    PubMed

    Huang, Qiao-Song; Yu, Zhao-Xian; Li, Jing; Chen, Chen

    2009-02-01

    In the present paper, the authors examined some oily core by microscopic infrared spectral imaging methods. Those methods can be classified in three modes, referred to as "transmission mode", "reflection mode" and "attenuated total reflection (ATR) mode". The observed oily core samples belong to siltstone. The samples were made of quartz (-20%), feldspar(-50%) and other rock (igneous rock 25%, metamorphic rocks 1%, sedimentary rock 4%); a little recrystallized calcite (-1%) was in the pore, and the argillaceous matter was distributed along the edge of a pore. The experimental work has been accomplished using SHIMADZU Model IRPrestige-21 Fourier transform infrared spectrophotometer plus AIM8800 infrared microscope. For IRPrestige-21, the spectral range is 7 800-350 cm(-1) spectral resolution is 1 cm(-1), and AIM8800 microscope with motorized stages has a resolution of 1 micrometer. The experiment was preformed at room temperature. In "transmission mode" infrared spectral imaging method, the spectral range was limited in wavenumbers greater than 2 000 cm(-1) because the base glass piece has strong light absorption. In contrast with "transmission mode", in "attenuated total reflection (ATR) mode", the depth of penetration into sample is very small (1-2 micrometer), then the absorbance value has nothing to do with base glass piece light absorption. In microscopic infrared transmission spectra, the experimental result shows that there are some strong absorption peaks at 2 866, 2 928, 3 618 and 2 515 cm(-1) respectively. The former two peaks correspond to methyl(methylene) symmetrical and unsymmetrical stretch vibration mode, respectively. The latter two peaks correspond to hydroxyl-stretch vibration mode and S-H, P-H chemical bond stretch vibration mode, respectively. In microscopic longwave infrared ATR spectra, there are other stronger absorption peaks at 1 400, 1 038 and 783 cm(i1)respectively, corresponding to methyl(methylene) widing vibration mode and optical mode

  2. [Microscopic infrared spectral imaging of oily core].

    PubMed

    Huang, Qiao-Song; Yu, Zhao-Xian; Li, Jing; Chen, Chen

    2009-02-01

    In the present paper, the authors examined some oily core by microscopic infrared spectral imaging methods. Those methods can be classified in three modes, referred to as "transmission mode", "reflection mode" and "attenuated total reflection (ATR) mode". The observed oily core samples belong to siltstone. The samples were made of quartz (-20%), feldspar(-50%) and other rock (igneous rock 25%, metamorphic rocks 1%, sedimentary rock 4%); a little recrystallized calcite (-1%) was in the pore, and the argillaceous matter was distributed along the edge of a pore. The experimental work has been accomplished using SHIMADZU Model IRPrestige-21 Fourier transform infrared spectrophotometer plus AIM8800 infrared microscope. For IRPrestige-21, the spectral range is 7 800-350 cm(-1) spectral resolution is 1 cm(-1), and AIM8800 microscope with motorized stages has a resolution of 1 micrometer. The experiment was preformed at room temperature. In "transmission mode" infrared spectral imaging method, the spectral range was limited in wavenumbers greater than 2 000 cm(-1) because the base glass piece has strong light absorption. In contrast with "transmission mode", in "attenuated total reflection (ATR) mode", the depth of penetration into sample is very small (1-2 micrometer), then the absorbance value has nothing to do with base glass piece light absorption. In microscopic infrared transmission spectra, the experimental result shows that there are some strong absorption peaks at 2 866, 2 928, 3 618 and 2 515 cm(-1) respectively. The former two peaks correspond to methyl(methylene) symmetrical and unsymmetrical stretch vibration mode, respectively. The latter two peaks correspond to hydroxyl-stretch vibration mode and S-H, P-H chemical bond stretch vibration mode, respectively. In microscopic longwave infrared ATR spectra, there are other stronger absorption peaks at 1 400, 1 038 and 783 cm(i1)respectively, corresponding to methyl(methylene) widing vibration mode and optical mode

  3. Clinical multiphoton and CARS microscopy

    NASA Astrophysics Data System (ADS)

    Breunig, H. G.; Weinigel, M.; Darvin, M. E.; Lademann, J.; König, K.

    2012-03-01

    We report on clinical CARS imaging of human skin in vivo with the certified hybrid multiphoton tomograph CARSDermaInspect. The CARS-DermaInspect provides simultaneous imaging of non-fluorescent intradermal lipid and water as well as imaging of two-photon excited fluorescence from intrinsic molecules. Two different excitation schemes for CARS imaging have been realized: In the first setup, a combination of fs oscillator and optical parametric oscillator provided fs-CARS pump and Stokes pulses, respectively. In the second setup a fs oscillator was combined with a photonic crystal fiber which provided a broadband spectrum. A spectral range out of the broadband-spectrum was selected and used for CARS excitation in combination with the residual fs-oscillator output. In both setups, in addition to CARS, single-beam excitation was used for imaging of two-photon excited fluorescence and second harmonic generation signals. Both CARS-excitation systems were successfully used for imaging of lipids inside the skin in vivo.

  4. The design of double electrostatic-lens optics for resonance enhanced multiphoton ionization and photoelectron imaging experiments

    SciTech Connect

    Qu, Zehua; Li, Chunsheng; Qin, Zhengbo E-mail: xfzheng@mail.ahnu.edu.cn; Zheng, Xianfeng E-mail: xfzheng@mail.ahnu.edu.cn; Yao, Guanxin; Zhang, Xianyi; Cui, Zhifeng

    2015-06-15

    Compared to single ion/electron-optics for velocity-map imaging, a double-focusing lens assembly designed not only allows for mapping velocity imaging of photoelectrons but also allows for investigating the vibrational structure of the intermediate states of neutral species in resonance enhanced multiphoton ionization (REMPI) spectra. In this presentation, in order to record REMPI and photoelectron spectra separately, we have constructed a compact photoelectron velocity-map imaging (VMI) apparatus combined with an opposite linear Wiley-Mclaren time-of-flight mass spectrometer (TOFMS). A mass resolution (m/Δm) of ∼1300 for TOFMS and electron energy resolution (ΔE/E) of 2.4% for VMI have been achieved upon three-photon ionization of Xe atom at 258.00 nm laser wavelength. As a benchmark, in combination of one-color (1 + 1) REMPI and photoelectron imaging of benzene via 6{sup 1} and 6{sup 1}1{sup 1} vibronic levels in the S{sub 1} state, the vibrational structures of the cation and photoelectron angular anisotropy are unraveled. In addition, two-color (1 + 1′) REMPI and photoelectron imaging of aniline was used to complete the accurate measurement of ionization potential (62 271 ± 3 cm{sup −1}). The results suggest that the apparatus is a powerful tool for studying photoionization dynamics in the photoelectron imaging using vibrational-state selected excitation to the intermediate states of neutrals based on REMPI technique.

  5. The design of double electrostatic-lens optics for resonance enhanced multiphoton ionization and photoelectron imaging experiments.

    PubMed

    Qu, Zehua; Li, Chunsheng; Qin, Zhengbo; Zheng, Xianfeng; Yao, Guanxin; Zhang, Xianyi; Cui, Zhifeng

    2015-06-01

    Compared to single ion/electron-optics for velocity-map imaging, a double-focusing lens assembly designed not only allows for mapping velocity imaging of photoelectrons but also allows for investigating the vibrational structure of the intermediate states of neutral species in resonance enhanced multiphoton ionization (REMPI) spectra. In this presentation, in order to record REMPI and photoelectron spectra separately, we have constructed a compact photoelectron velocity-map imaging (VMI) apparatus combined with an opposite linear Wiley-Mclaren time-of-flight mass spectrometer (TOFMS). A mass resolution (m/Δm) of ∼1300 for TOFMS and electron energy resolution (ΔE/E) of 2.4% for VMI have been achieved upon three-photon ionization of Xe atom at 258.00 nm laser wavelength. As a benchmark, in combination of one-color (1 + 1) REMPI and photoelectron imaging of benzene via 6(1) and 6(1)1(1) vibronic levels in the S1 state, the vibrational structures of the cation and photoelectron angular anisotropy are unraveled. In addition, two-color (1 + 1') REMPI and photoelectron imaging of aniline was used to complete the accurate measurement of ionization potential (62,271 ± 3 cm(-1)). The results suggest that the apparatus is a powerful tool for studying photoionization dynamics in the photoelectron imaging using vibrational-state selected excitation to the intermediate states of neutrals based on REMPI technique. PMID:26133827

  6. The design of double electrostatic-lens optics for resonance enhanced multiphoton ionization and photoelectron imaging experiments

    NASA Astrophysics Data System (ADS)

    Qu, Zehua; Li, Chunsheng; Qin, Zhengbo; Zheng, Xianfeng; Yao, Guanxin; Zhang, Xianyi; Cui, Zhifeng

    2015-06-01

    Compared to single ion/electron-optics for velocity-map imaging, a double-focusing lens assembly designed not only allows for mapping velocity imaging of photoelectrons but also allows for investigating the vibrational structure of the intermediate states of neutral species in resonance enhanced multiphoton ionization (REMPI) spectra. In this presentation, in order to record REMPI and photoelectron spectra separately, we have constructed a compact photoelectron velocity-map imaging (VMI) apparatus combined with an opposite linear Wiley-Mclaren time-of-flight mass spectrometer (TOFMS). A mass resolution (m/Δm) of ˜1300 for TOFMS and electron energy resolution (ΔE/E) of 2.4% for VMI have been achieved upon three-photon ionization of Xe atom at 258.00 nm laser wavelength. As a benchmark, in combination of one-color (1 + 1) REMPI and photoelectron imaging of benzene via 61 and 6111 vibronic levels in the S1 state, the vibrational structures of the cation and photoelectron angular anisotropy are unraveled. In addition, two-color (1 + 1') REMPI and photoelectron imaging of aniline was used to complete the accurate measurement of ionization potential (62 271 ± 3 cm-1). The results suggest that the apparatus is a powerful tool for studying photoionization dynamics in the photoelectron imaging using vibrational-state selected excitation to the intermediate states of neutrals based on REMPI technique.

  7. In vivo, label-free, three-dimensional quantitative imaging of liver surface using multi-photon microscopy

    NASA Astrophysics Data System (ADS)

    Zhuo, Shuangmu; Yan, Jie; Kang, Yuzhan; Xu, Shuoyu; Peng, Qiwen; So, Peter T. C.; Yu, Hanry

    2014-07-01

    Various structural features on the liver surface reflect functional changes in the liver. The visualization of these surface features with molecular specificity is of particular relevance to understanding the physiology and diseases of the liver. Using multi-photon microscopy (MPM), we have developed a label-free, three-dimensional quantitative and sensitive method to visualize various structural features of liver surface in living rat. MPM could quantitatively image the microstructural features of liver surface with respect to the sinuosity of collagen fiber, the elastic fiber structure, the ratio between elastin and collagen, collagen content, and the metabolic state of the hepatocytes that are correlative with the pathophysiologically induced changes in the regions of interest. This study highlights the potential of this technique as a useful tool for pathophysiological studies and possible diagnosis of the liver diseases with further development.

  8. In vivo, label-free, three-dimensional quantitative imaging of liver surface using multi-photon microscopy

    SciTech Connect

    Zhuo, Shuangmu E-mail: hanry-yu@nuhs.edu.sg; Yan, Jie; Kang, Yuzhan; Peng, Qiwen; and others

    2014-07-14

    Various structural features on the liver surface reflect functional changes in the liver. The visualization of these surface features with molecular specificity is of particular relevance to understanding the physiology and diseases of the liver. Using multi-photon microscopy (MPM), we have developed a label-free, three-dimensional quantitative and sensitive method to visualize various structural features of liver surface in living rat. MPM could quantitatively image the microstructural features of liver surface with respect to the sinuosity of collagen fiber, the elastic fiber structure, the ratio between elastin and collagen, collagen content, and the metabolic state of the hepatocytes that are correlative with the pathophysiologically induced changes in the regions of interest. This study highlights the potential of this technique as a useful tool for pathophysiological studies and possible diagnosis of the liver diseases with further development.

  9. Characterizing liver capsule microstructure via in situ bulge test coupled with multiphoton imaging.

    PubMed

    Jayyosi, C; Coret, M; Bruyère-Garnier, K

    2016-02-01

    The characterization of biological tissue at the microscopic scale is the starting point of many applications in tissue engineering and especially in the development of structurally based constitutive models. In the present study, focus is made on the liver capsule, the membrane encompassing hepatic parenchyma, which takes a huge part in liver mechanical properties. An in situ bulge test experiment under a multiphoton microscope has been developed to assess the microstructure changes that arise with biaxial loading. Multiphoton microscopy allows to observe the elastin and collagen fiber networks simultaneously. Thus a description of the microstructure organization of the capsule is given, characterizing the shapes, geometry and arrangement of fibers. The orientation of fibers is calculated and orientation distribution evolution with loading is given, in the case of an equibiaxial and two non equibiaxial loadings, thanks to a circular and elliptic set up of the bulge test. The local strain fields have also been computed, by the mean of a photobleaching grid, to get an idea of what the liver capsule might experience when subjected to internal pressure. Results show that strain fields present some heterogeneity due to anisotropy. Reorientation occurs in non equibiaxial loadings and involves fibers layers from the inner to the outer surface as expected. Although there is a fiber network rearrangement to accommodate with loading in the case of equibiaxial loading, there is no significant reorientation of the main fibers direction of the different layers.

  10. Ultrafast Imaging using Spectral Resonance Modulation

    NASA Astrophysics Data System (ADS)

    Huang, Eric; Ma, Qian; Liu, Zhaowei

    2016-04-01

    CCD cameras are ubiquitous in research labs, industry, and hospitals for a huge variety of applications, but there are many dynamic processes in nature that unfold too quickly to be captured. Although tradeoffs can be made between exposure time, sensitivity, and area of interest, ultimately the speed limit of a CCD camera is constrained by the electronic readout rate of the sensors. One potential way to improve the imaging speed is with compressive sensing (CS), a technique that allows for a reduction in the number of measurements needed to record an image. However, most CS imaging methods require spatial light modulators (SLMs), which are subject to mechanical speed limitations. Here, we demonstrate an etalon array based SLM without any moving elements that is unconstrained by either mechanical or electronic speed limitations. This novel spectral resonance modulator (SRM) shows great potential in an ultrafast compressive single pixel camera.

  11. Ultrafast Imaging using Spectral Resonance Modulation.

    PubMed

    Huang, Eric; Ma, Qian; Liu, Zhaowei

    2016-01-01

    CCD cameras are ubiquitous in research labs, industry, and hospitals for a huge variety of applications, but there are many dynamic processes in nature that unfold too quickly to be captured. Although tradeoffs can be made between exposure time, sensitivity, and area of interest, ultimately the speed limit of a CCD camera is constrained by the electronic readout rate of the sensors. One potential way to improve the imaging speed is with compressive sensing (CS), a technique that allows for a reduction in the number of measurements needed to record an image. However, most CS imaging methods require spatial light modulators (SLMs), which are subject to mechanical speed limitations. Here, we demonstrate an etalon array based SLM without any moving elements that is unconstrained by either mechanical or electronic speed limitations. This novel spectral resonance modulator (SRM) shows great potential in an ultrafast compressive single pixel camera. PMID:27122101

  12. Ultrafast Imaging using Spectral Resonance Modulation

    PubMed Central

    Huang, Eric; Ma, Qian; Liu, Zhaowei

    2016-01-01

    CCD cameras are ubiquitous in research labs, industry, and hospitals for a huge variety of applications, but there are many dynamic processes in nature that unfold too quickly to be captured. Although tradeoffs can be made between exposure time, sensitivity, and area of interest, ultimately the speed limit of a CCD camera is constrained by the electronic readout rate of the sensors. One potential way to improve the imaging speed is with compressive sensing (CS), a technique that allows for a reduction in the number of measurements needed to record an image. However, most CS imaging methods require spatial light modulators (SLMs), which are subject to mechanical speed limitations. Here, we demonstrate an etalon array based SLM without any moving elements that is unconstrained by either mechanical or electronic speed limitations. This novel spectral resonance modulator (SRM) shows great potential in an ultrafast compressive single pixel camera. PMID:27122101

  13. Distinguishing human normal or cancerous esophagus tissue ex vivo using multiphoton microscopy

    NASA Astrophysics Data System (ADS)

    Liu, N. R.; Chen, G. N.; Wu, S. S.; Chen, R.

    2014-02-01

    Application of multiphoton microscopy (MPM) to clinical cancer research has greatly developed over the last few years. In this paper, we mainly focus on two-photon excitation fluorescence (TPEF) and second harmonic generation (SHG) for investigating esophageal cancer. We chiefly discuss the SHG/TPEF image and spectral characteristics of normal and cancerous esophagus submucosa with the combined multi-channel imaging mode and Lambda mode of a multiphoton microscope (LSM 510 META). Great differences can be detected, such as collagen content and morphology, glandular-shaped cancer cells, TPEF/SHG intensity ratio, and so on, which demonstrate that the multiphoton imaging technique has the potential ability for minimally-invasive early cancer diagnosis.

  14. Influence of Vacuum Cooling on Escherichia coli O157:H7 Infiltration in Fresh Leafy Greens via a Multiphoton-Imaging Approach.

    PubMed

    Vonasek, Erica; Nitin, Nitin

    2015-10-16

    Microbial pathogen infiltration in fresh leafy greens is a significant food safety risk factor. In various postharvest operations, vacuum cooling is a critical process for maintaining the quality of fresh produce. The overall goal of this study was to evaluate the risk of vacuum cooling-induced infiltration of Escherichia coli O157:H7 into lettuce using multiphoton microscopy. Multiphoton imaging was chosen as the method to locate E. coli O157:H7 within an intact lettuce leaf due to its high spatial resolution, low background fluorescence, and near-infrared (NIR) excitation source compared to those of conventional confocal microscopy. The variables vacuum cooling, surface moisture, and leaf side were evaluated in a three-way factorial study with E. coli O157:H7 on lettuce. A total of 188 image stacks were collected. The images were analyzed for E. coli O157:H7 association with stomata and E. coli O157:H7 infiltration. The quantitative imaging data were statistically analyzed using analysis of variance (ANOVA). The results indicate that the low-moisture condition led to an increased risk of microbial association with stomata (P < 0.05). Additionally, the interaction between vacuum cooling levels and moisture levels led to an increased risk of infiltration (P < 0.05). This study also demonstrates the potential of multiphoton imaging for improving sensitivity and resolution of imaging-based measurements of microbial interactions with intact leaf structures, including infiltration.

  15. Influence of Vacuum Cooling on Escherichia coli O157:H7 Infiltration in Fresh Leafy Greens via a Multiphoton-Imaging Approach.

    PubMed

    Vonasek, Erica; Nitin, Nitin

    2016-01-01

    Microbial pathogen infiltration in fresh leafy greens is a significant food safety risk factor. In various postharvest operations, vacuum cooling is a critical process for maintaining the quality of fresh produce. The overall goal of this study was to evaluate the risk of vacuum cooling-induced infiltration of Escherichia coli O157:H7 into lettuce using multiphoton microscopy. Multiphoton imaging was chosen as the method to locate E. coli O157:H7 within an intact lettuce leaf due to its high spatial resolution, low background fluorescence, and near-infrared (NIR) excitation source compared to those of conventional confocal microscopy. The variables vacuum cooling, surface moisture, and leaf side were evaluated in a three-way factorial study with E. coli O157:H7 on lettuce. A total of 188 image stacks were collected. The images were analyzed for E. coli O157:H7 association with stomata and E. coli O157:H7 infiltration. The quantitative imaging data were statistically analyzed using analysis of variance (ANOVA). The results indicate that the low-moisture condition led to an increased risk of microbial association with stomata (P < 0.05). Additionally, the interaction between vacuum cooling levels and moisture levels led to an increased risk of infiltration (P < 0.05). This study also demonstrates the potential of multiphoton imaging for improving sensitivity and resolution of imaging-based measurements of microbial interactions with intact leaf structures, including infiltration. PMID:26475109

  16. In-vivo imaging of psoriatic lesions with polarization multispectral dermoscopy and multiphoton microscopy

    PubMed Central

    Kapsokalyvas, Dimitrios; Cicchi, Riccardo; Bruscino, Nicola; Alfieri, Domenico; Prignano, Francesca; Massi, Daniela; Lotti, Torello; Pavone, Francesco S.

    2014-01-01

    Psoriasis is a skin autoimmune disease characterized by hyperkeratosis, hyperproliferation of the epidermis and dilatation of dermal papillary blood vessels. Healthy skin (5 volunteers) and psoriatic lesions (3 patients) were visualized in vivo, with high contrast and resolution, with a Polarization Multispectral Dermoscope and a Multiphoton Microscope. Psoriatic features were identified and quantified. The effective diameter of the superficial blood vessels was measured at 35.2 ± 7.2 μm and the elongated dermal papillae had an effective diameter of 64.2 ± 22.6 μm. The methodologies developed could be employed for quantitative diagnostic purposes and furthermore serve as a monitoring method of the effect of personalized treatments. PMID:25071974

  17. All-near-infrared multiphoton microscopy interrogates intact tissues at deeper imaging depths than conventional single- and two-photon near-infrared excitation microscopes.

    PubMed

    Sarder, Pinaki; Yazdanfar, Siavash; Akers, Walter J; Tang, Rui; Sudlow, Gail P; Egbulefu, Christopher; Achilefu, Samuel

    2013-10-01

    The era of molecular medicine has ushered in the development of microscopic methods that can report molecular processes in thick tissues with high spatial resolution. A commonality in deep-tissue microscopy is the use of near-infrared (NIR) lasers with single- or multiphoton excitations. However, the relationship between different NIR excitation microscopic techniques and the imaging depths in tissue has not been established. We compared such depth limits for three NIR excitation techniques: NIR single-photon confocal microscopy (NIR SPCM), NIR multiphoton excitation with visible detection (NIR/VIS MPM), and all-NIR multiphoton excitation with NIR detection (NIR/NIR MPM). Homologous cyanine dyes provided the fluorescence. Intact kidneys were harvested after administration of kidney-clearing cyanine dyes in mice. NIR SPCM and NIR/VIS MPM achieved similar maximum imaging depth of ∼100 μm. The NIR/NIR MPM enabled greater than fivefold imaging depth (>500 μm) using the harvested kidneys. Although the NIR/NIR MPM used 1550-nm excitation where water absorption is relatively high, cell viability and histology studies demonstrate that the laser did not induce photothermal damage at the low laser powers used for the kidney imaging. This study provides guidance on the imaging depth capabilities of NIR excitation-based microscopic techniques and reveals the potential to multiplex information using these platforms.

  18. HYMOSS signal processing for pushbroom spectral imaging

    NASA Technical Reports Server (NTRS)

    Ludwig, David E.

    1991-01-01

    The objective of the Pushbroom Spectral Imaging Program was to develop on-focal plane electronics which compensate for detector array non-uniformities. The approach taken was to implement a simple two point calibration algorithm on focal plane which allows for offset and linear gain correction. The key on focal plane features which made this technique feasible was the use of a high quality transimpedance amplifier (TIA) and an analog-to-digital converter for each detector channel. Gain compensation is accomplished by varying the feedback capacitance of the integrate and dump TIA. Offset correction is performed by storing offsets in a special on focal plane offset register and digitally subtracting the offsets from the readout data during the multiplexing operation. A custom integrated circuit was designed, fabricated, and tested on this program which proved that nonuniformity compensated, analog-to-digital converting circuits may be used to read out infrared detectors. Irvine Sensors Corporation (ISC) successfully demonstrated the following innovative on-focal-plane functions that allow for correction of detector non-uniformities. Most of the circuit functions demonstrated on this program are finding their way onto future IC's because of their impact on reduced downstream processing, increased focal plane performance, simplified focal plane control, reduced number of dewar connections, as well as the noise immunity of a digital interface dewar. The potential commercial applications for this integrated circuit are primarily in imaging systems. These imaging systems may be used for: security monitoring systems, manufacturing process monitoring, robotics, and for spectral imaging when used in analytical instrumentation.

  19. HYMOSS signal processing for pushbroom spectral imaging

    NASA Astrophysics Data System (ADS)

    Ludwig, David E.

    1991-06-01

    The objective of the Pushbroom Spectral Imaging Program was to develop on-focal plane electronics which compensate for detector array non-uniformities. The approach taken was to implement a simple two point calibration algorithm on focal plane which allows for offset and linear gain correction. The key on focal plane features which made this technique feasible was the use of a high quality transimpedance amplifier (TIA) and an analog-to-digital converter for each detector channel. Gain compensation is accomplished by varying the feedback capacitance of the integrate and dump TIA. Offset correction is performed by storing offsets in a special on focal plane offset register and digitally subtracting the offsets from the readout data during the multiplexing operation. A custom integrated circuit was designed, fabricated, and tested on this program which proved that nonuniformity compensated, analog-to-digital converting circuits may be used to read out infrared detectors. Irvine Sensors Corporation (ISC) successfully demonstrated the following innovative on-focal-plane functions that allow for correction of detector non-uniformities. Most of the circuit functions demonstrated on this program are finding their way onto future IC's because of their impact on reduced downstream processing, increased focal plane performance, simplified focal plane control, reduced number of dewar connections, as well as the noise immunity of a digital interface dewar. The potential commercial applications for this integrated circuit are primarily in imaging systems. These imaging systems may be used for: security monitoring systems, manufacturing process monitoring, robotics, and for spectral imaging when used in analytical instrumentation.

  20. Multiphoton microspectroscopy of biological specimens

    NASA Astrophysics Data System (ADS)

    Lin, Bai-Ling; Kao, Fu-Jen; Cheng, Ping C.; Sun, Chi-Kuang; Chen, RangWu; Wang, YiMin; Chen, JianCheng; Wang, Yung-Shun; Liu, Tzu-Ming; Huang, Mao-Kuo

    2000-07-01

    The non-linear nature of multi-photon fluorescence excitation restricts the fluorescing volume to the vicinity of the focal point. As a result, the technology has the capacity for micro- spectroscopy of biological specimen at high spatial resolution. Chloroplasts in mesophyll protoplast of Arabidopsis thaliana and maize stem sections were used to demonstrate the feasibility of multi-photon fluorescence micro-spectroscopy at subcellular compartments. Time-lapse spectral recording provides a means for studying the response of cell organelles to high intensity illumination.

  1. Assessment of liver steatosis and fibrosis in rats using integrated coherent anti-Stokes Raman scattering and multiphoton imaging technique

    NASA Astrophysics Data System (ADS)

    Lin, Jian; Lu, Fake; Zheng, Wei; Xu, Shuoyu; Tai, Dean; Yu, Hanry; Huang, Zhiwei

    2011-11-01

    We report the implementation of a unique integrated coherent anti-Stokes Raman scattering (CARS), second-harmonic generation (SHG), and two-photon excitation fluorescence (TPEF) microscopy imaging technique developed for label-free monitoring of the progression of liver steatosis and fibrosis generated in a bile duct ligation (BDL) rat model. Among the 21 adult rats used in this study, 18 rats were performed with BDL surgery and sacrificed each week from weeks 1 to 6 (n = 3 per week), respectively; whereas 3 rats as control were sacrificed at week 0. Colocalized imaging of the aggregated hepatic fats, collagen fibrils, and hepatocyte morphologies in liver tissue is realized by using the integrated CARS, SHG, and TPEF technique. The results show that there are significant accumulations of hepatic lipid droplets and collagen fibrils associated with severe hepatocyte necrosis in BDL rat liver as compared to a normal liver tissue. The volume of normal hepatocytes keeps decreasing and the fiber collagen content in BDL rat liver follows a growing trend until week 6; whereas the hepatic fat content reaches a maximum in week 4 and then appears to stop growing in week 6, indicating that liver steatosis and fibrosis induced in a BDL rat liver model may develop at different rates. This work demonstrates that the integrated CARS and multiphoton microscopy imaging technique has the potential to provide an effective means for early diagnosis and detection of liver steatosis and fibrosis without labeling.

  2. Detection limits with spectral differential imaging data

    NASA Astrophysics Data System (ADS)

    Rameau, J.; Chauvin, G.; Lagrange, A.-M.; Maire, A.-L.; Boccaletti, A.; Bonnefoy, M.

    2015-09-01

    Context. Direct imaging of exoplanets is polluted by speckle noise that severely limits the achievable contrast. Angular and spectral differential imaging have been proposed to make use of the temporal and chromatic properties of the speckles. Both modes, associated with extreme adaptive-optics and coronagraphy, are at the core of the new generation of planet imagers SPHERE and GPI. Aims: We aim to illustrate and characterize the impact of the SDI and SDI+ADI (ASDI) data reduction on the detection of giant planets. We also propose an unbiased method to derive the detection limits from SDI/ASDI data. Methods: Observations of AB Dor B and β Pictoris made with VLT/NaCo were used to simulate and quantify the effects of SDI and ASDI. The novel method is compared to the traditional injection of artificial point sources. Results: The SDI reduction process creates a typical radial positive-negative pattern of any point-source. Its characteristics and its self-subtraction depend on the separation, but also on the spectral properties of the object. This work demonstrates that the self-subtraction cannot be reduced to a simple geometric effect. As a consequence, the detection performances of SDI observations cannot be expressed as a contrast in magnitude with the central star without the knowledge of the spectral properties of detectable companions. In addition, the residual noise cannot be converted into contrast and physical characteristics (mass, temperature) by standard calibration of flux losses. The proposed method takes the SDI bias into account to derive detection limits without the cost of massively injecting artificial sources into the data. Finally, the sensitivity of ASDI observations can be measured only with a control parameter on the algorithms that controls the minimum rotation that is necessary to build the reference image. Based on observations collected at the European Organization for Astronomical Research in the Southern Hemisphere, Chile, ESO : 60.A

  3. Multiphoton excitation microscopy of in vivo human skin. Functional and morphological optical biopsy based on three-dimensional imaging, lifetime measurements and fluorescence spectroscopy.

    PubMed

    Masters, B R; So, P T; Gratton, E

    1998-02-01

    Two-photon excitation microscopy has the potential as an effective, noninvasive, diagnostic tool for in vivo examination of human deep tissue structure at the subcellular level. By using infrared photons as the excitation source in two-photon microscopy, a significant improvement in penetration depth can be achieved because of the much lower tissue scattering and absorption coefficients in the infrared wavelengths. Two-photon absorption occurs primarily at the focal point and provides the physical basis for optical sectioning. Multiphoton excitation microscopy at 730 nm was used to image in vivo human skin autofluorescence from the surface to a depth of about 200 microns. The spectroscopic data suggest that reduced pyridine nucleotides, NAD(P)H, are the primary source of the skin autofluorescence using 730 nm excitation. This study demonstrates the use of multiphoton excitation microscopy for functional imaging of the metabolic states of in vivo human skin cells and provides a functional and morphological optical biopsy.

  4. Multiphoton microscopy, fluorescence lifetime imaging and optical spectroscopy for the diagnosis of neoplasia

    NASA Astrophysics Data System (ADS)

    Skala, Melissa Caroline

    2007-12-01

    Cancer morbidity and mortality is greatly reduced when the disease is diagnosed and treated early in its development. Tissue biopsies are the gold standard for cancer diagnosis, and an accurate diagnosis requires a biopsy from the malignant portion of an organ. Light, guided through a fiber optic probe, could be used to inspect regions of interest and provide real-time feedback to determine the optimal tissue site for biopsy. This approach could increase the diagnostic accuracy of current biopsy procedures. The studies in this thesis have characterized changes in tissue optical signals with carcinogenesis, increasing our understanding of the sensitivity of optical techniques for cancer detection. All in vivo studies were conducted on the dimethylbenz[alpha]anthracene treated hamster cheek pouch model of epithelial carcinogenesis. Multiphoton microscopy studies in the near infrared wavelength region quantified changes in tissue morphology and fluorescence with carcinogenesis in vivo. Statistically significant morphological changes with precancer included increased epithelial thickness, loss of stratification in the epithelium, and increased nuclear diameter. Fluorescence changes included a statistically significant decrease in the epithelial fluorescence intensity per voxel at 780 nm excitation, a decrease in the fluorescence lifetime of protein-bound nicotinamide adenine dinucleotide (NADH, an electron donor in oxidative phosphorylation), and an increase in the fluorescence lifetime of protein-bound flavin adenine dinucleotide (FAD, an electron acceptor in oxidative phosphorylation) with precancer. The redox ratio (fluorescence intensity of FAD/NADH, a measure of the cellular oxidation-reduction state) did not significantly change with precancer. Cell culture experiments (MCF10A cells) indicated that the decrease in protein-bound NADH with precancer could be due to increased levels of glycolysis. Point measurements of diffuse reflectance and fluorescence spectra in

  5. Spectral imaging of neurosurgical target tissues through operation microscope

    NASA Astrophysics Data System (ADS)

    Antikainen, Jukka; von Und Zu Fraunberg, Mikael; Orava, Joni; Jaaskelainen, Juha E.; Hauta-Kasari, Markku

    2011-11-01

    It has been noticed that spectral information can be used for analyzing and separating different biological tissues. However, most of the studies for spectral image acquisitions are mainly done in vitro. Usually the main restrictions for in vivo measurements are the size or the weight of the spectral camera. If the camera weights too much, the surgery microscope cannot be stabilized. If the size of the camera is too big, it will disturb the surgeon or even risk the safety of the patient. The main goal of this study was to develop an independent spectral imaging device which can be used for collecting spectral information from the neurosurgeries without any previously described restrictions. Size of the imaging system is small enough not to disturb the surgeon during the surgery. The developed spectral imaging system is used for collecting a spectral database which can be used for the future imaging systems.

  6. Intravital multiphoton imaging of the selective uptake of water-dispersible quantum dots into sinusoidal liver cells.

    PubMed

    Liang, Xiaowen; Grice, Jeffrey E; Zhu, Yian; Liu, David; Sanchez, Washington Y; Li, Zhen; Crawford, Darrell H G; Le Couteur, David G; Cogger, Victoria C; Liu, Xin; Xu, Zhi Ping; Roberts, Michael S

    2015-04-01

    Although many studies reporting the organ-level biodistribution of nanoparticles (NPs) in animals, very few have addressed the fate of NPs in organs at the cellular level. The liver appears to be the main organ for accumulation of NPs after intravenous injection. In this study, for the first time, the in vivo spatiotemporal disposition of recently developed mercaptosuccinic acid (MSA)-capped cadmium telluride/cadmium sulfide (CdTe/CdS) quantum dots (QDs) is explored in rat liver using multiphoton microscopy (MPM) coupled with fluorescence lifetime imaging (FLIM), with subcellular resolution (∼1 μm). With high fluorescence efficiency and largely improved stability in the biological environment, these QDs show a distinct distribution pattern in the liver compared to organic dyes, rhodamine 123 and fluorescein. After intravenous injection, fluorescent molecules are taken up by hepatocytes and excreted into the bile, while negatively charged QDs are retained in the sinusoids and selectively taken up by sinusoidal cells (Kupffer cells and liver sinusoidal endothelial cells), but not by hepatocytes within 3 h. The results could help design NPs targeting the specific types of liver cells and choose the fluorescent markers for appropriate cellular imaging.

  7. Spectral Camera based on Ghost Imaging via Sparsity Constraints

    NASA Astrophysics Data System (ADS)

    Liu, Zhentao; Tan, Shiyu; Wu, Jianrong; Li, Enrong; Shen, Xia; Han, Shensheng

    2016-05-01

    The image information acquisition ability of a conventional camera is usually much lower than the Shannon Limit since it does not make use of the correlation between pixels of image data. Applying a random phase modulator to code the spectral images and combining with compressive sensing (CS) theory, a spectral camera based on true thermal light ghost imaging via sparsity constraints (GISC spectral camera) is proposed and demonstrated experimentally. GISC spectral camera can acquire the information at a rate significantly below the Nyquist rate, and the resolution of the cells in the three-dimensional (3D) spectral images data-cube can be achieved with a two-dimensional (2D) detector in a single exposure. For the first time, GISC spectral camera opens the way of approaching the Shannon Limit determined by Information Theory in optical imaging instruments.

  8. Spectral Camera based on Ghost Imaging via Sparsity Constraints.

    PubMed

    Liu, Zhentao; Tan, Shiyu; Wu, Jianrong; Li, Enrong; Shen, Xia; Han, Shensheng

    2016-05-16

    The image information acquisition ability of a conventional camera is usually much lower than the Shannon Limit since it does not make use of the correlation between pixels of image data. Applying a random phase modulator to code the spectral images and combining with compressive sensing (CS) theory, a spectral camera based on true thermal light ghost imaging via sparsity constraints (GISC spectral camera) is proposed and demonstrated experimentally. GISC spectral camera can acquire the information at a rate significantly below the Nyquist rate, and the resolution of the cells in the three-dimensional (3D) spectral images data-cube can be achieved with a two-dimensional (2D) detector in a single exposure. For the first time, GISC spectral camera opens the way of approaching the Shannon Limit determined by Information Theory in optical imaging instruments.

  9. Spectral Camera based on Ghost Imaging via Sparsity Constraints

    PubMed Central

    Liu, Zhentao; Tan, Shiyu; Wu, Jianrong; Li, Enrong; Shen, Xia; Han, Shensheng

    2016-01-01

    The image information acquisition ability of a conventional camera is usually much lower than the Shannon Limit since it does not make use of the correlation between pixels of image data. Applying a random phase modulator to code the spectral images and combining with compressive sensing (CS) theory, a spectral camera based on true thermal light ghost imaging via sparsity constraints (GISC spectral camera) is proposed and demonstrated experimentally. GISC spectral camera can acquire the information at a rate significantly below the Nyquist rate, and the resolution of the cells in the three-dimensional (3D) spectral images data-cube can be achieved with a two-dimensional (2D) detector in a single exposure. For the first time, GISC spectral camera opens the way of approaching the Shannon Limit determined by Information Theory in optical imaging instruments. PMID:27180619

  10. Spectral embedded hashing for scalable image retrieval.

    PubMed

    Chen, Lin; Xu, Dong; Tsang, Ivor Wai-Hung; Li, Xuelong

    2014-07-01

    We propose a new graph based hashing method called spectral embedded hashing (SEH) for large-scale image retrieval. We first introduce a new regularizer into the objective function of the recent work spectral hashing to control the mismatch between the resultant hamming embedding and the low-dimensional data representation, which is obtained by using a linear regression function. This linear regression function can be employed to effectively handle the out-of-sample data, and the introduction of the new regularizer makes SEH better cope with the data sampled from a nonlinear manifold. Considering that SEH cannot efficiently cope with the high dimensional data, we further extend SEH to kernel SEH (KSEH) to improve the efficiency and effectiveness, in which a nonlinear regression function can also be employed to obtain the low dimensional data representation. We also develop a new method to efficiently solve the approximate solution for the eigenvalue decomposition problem in SEH and KSEH. Moreover, we show that some existing hashing methods are special cases of our KSEH. Our comprehensive experiments on CIFAR, Tiny-580K, NUS-WIDE, and Caltech-256 datasets clearly demonstrate the effectiveness of our methods.

  11. Detection of latent fingerprints by near-infrared spectral imaging

    NASA Astrophysics Data System (ADS)

    Huang, Wei; Dai, Yong

    2014-05-01

    Spectral imaging technology research is becoming more extensive in the field of examination of material evidence. Near-Infrared spectral imaging technology is an important part of the full spectrum of imaging technology. This paper finished the experiment contents of the Near-Infrared spectrum imaging method and image acquisition system Near-Infrared spectral imaging technology. The experiment of Near-Infrared spectral imaging method obtains the image set of the Near-Infrared spectrum, and formats a pseudo-color images to show the potential traces successfully by processing the set of spectral images; Near-Infrared spectral imaging technology explores the technology method of obtaining the image set of Near-Infrared spectrometer and image acquisition system, and extensive access to the Near-Infrared spectrum information of latent blood, stamp and smear fingerprints on common objects, and study the characteristics of the Near-Infrared spectrum. Near-Infrared spectroscopic imaging experiments explores a wide variety of Near-Infrared reflectance spectra of the object material curve and its Near-Infrared spectrum of imaging modalities, can not only gives a reference for choosing Near-Infrared wavelength to show the object surface potential traces of substances, but also gives important data for the Near-Infrared spectrum of imaging technology development.

  12. A new method using multiphoton imaging and morphometric analysis for differentiating chromophobe renal cell carcinoma and oncocytoma kidney tumors

    NASA Astrophysics Data System (ADS)

    Wu, Binlin; Mukherjee, Sushmita; Jain, Manu

    2016-03-01

    Distinguishing chromophobe renal cell carcinoma (chRCC) from oncocytoma on hematoxylin and eosin images may be difficult and require time-consuming ancillary procedures. Multiphoton microscopy (MPM), an optical imaging modality, was used to rapidly generate sub-cellular histological resolution images from formalin-fixed unstained tissue sections from chRCC and oncocytoma.Tissues were excited using 780nm wavelength and emission signals (including second harmonic generation and autofluorescence) were collected in different channels between 390 nm and 650 nm. Granular structure in the cell cytoplasm was observed in both chRCC and oncocytoma. Quantitative morphometric analysis was conducted to distinguish chRCC and oncocytoma. To perform the analysis, cytoplasm and granules in tumor cells were segmented from the images. Their area and fluorescence intensity were found in different channels. Multiple features were measured to quantify the morphological and fluorescence properties. Linear support vector machine (SVM) was used for classification. Re-substitution validation, cross validation and receiver operating characteristic (ROC) curve were implemented to evaluate the efficacy of the SVM classifier. A wrapper feature algorithm was used to select the optimal features which provided the best predictive performance in separating the two tissue types (classes). Statistical measures such as sensitivity, specificity, accuracy and area under curve (AUC) of ROC were calculated to evaluate the efficacy of the classification. Over 80% accuracy was achieved as the predictive performance. This method, if validated on a larger and more diverse sample set, may serve as an automated rapid diagnostic tool to differentiate between chRCC and oncocytoma. An advantage of such automated methods are that they are free from investigator bias and variability.

  13. Multiphoton cryo microscope with sample temperature control

    NASA Astrophysics Data System (ADS)

    Breunig, H. G.; Uchugonova, A.; König, K.

    2013-02-01

    We present a multiphoton microscope system which combines the advantages of multiphoton imaging with precise control of the sample temperature. The microscope provides online insight in temperature-induced changes and effects in plant tissue and animal cells with subcellular resolution during cooling and thawing processes. Image contrast is based on multiphoton fluorescence intensity or fluorescence lifetime in the range from liquid nitrogen temperature up to +600°C. In addition, micro spectra from the imaged regions can be recorded. We present measurement results from plant leaf samples as well as Chinese hamster ovary cells.

  14. High spectral resolution image of Barnacle Bill

    NASA Technical Reports Server (NTRS)

    1997-01-01

    The rover Sojourner's first target for measurement by the Alpha-Proton-Xray Spectrometer (APXS) was the rock named Barnacle Bill, located close to the ramp down which the rover made its egress from the lander. The full spectral capability of the Imager for Mars Pathfinder (IMP), consisting of 13 wavelength filters, was used to characterize the rock's surface. The measured area is relatively dark, and is shown in blue. Nearby on the rock surface, soil material is trapped in pits (shown in red).

    Mars Pathfinder is the second in NASA's Discovery program of low-cost spacecraft with highly focused science goals. The Jet Propulsion Laboratory, Pasadena, CA, developed and manages the Mars Pathfinder mission for NASA's Office of Space Science, Washington, D.C. The Imager for Mars Pathfinder (IMP) was developed by the University of Arizona Lunar and Planetary Laboratory under contract to JPL. Peter Smith is the Principal Investigator. JPL is an operating division of the California Institute of Technology (Caltech).

  15. Long term intravital multiphoton microscopy imaging of immune cells in healthy and diseased liver using CXCR6.Gfp reporter mice.

    PubMed

    Heymann, Felix; Niemietz, Patricia M; Peusquens, Julia; Ergen, Can; Kohlhepp, Marlene; Mossanen, Jana C; Schneider, Carlo; Vogt, Michael; Tolba, Rene H; Trautwein, Christian; Martin, Christian; Tacke, Frank

    2015-01-01

    Liver inflammation as a response to injury is a highly dynamic process involving the infiltration of distinct subtypes of leukocytes including monocytes, neutrophils, T cell subsets, B cells, natural killer (NK) and NKT cells. Intravital microscopy of the liver for monitoring immune cell migration is particularly challenging due to the high requirements regarding sample preparation and fixation, optical resolution and long-term animal survival. Yet, the dynamics of inflammatory processes as well as cellular interaction studies could provide critical information to better understand the initiation, progression and regression of inflammatory liver disease. Therefore, a highly sensitive and reliable method was established to study migration and cell-cell-interactions of different immune cells in mouse liver over long periods (about 6 hr) by intravital two-photon laser scanning microscopy (TPLSM) in combination with intensive care monitoring. The method provided includes a gentle preparation and stable fixation of the liver with minimal perturbation of the organ; long term intravital imaging using multicolor multiphoton microscopy with virtually no photobleaching or phototoxic effects over a time period of up to 6 hr, allowing tracking of specific leukocyte subsets; and stable imaging conditions due to extensive monitoring of mouse vital parameters and stabilization of circulation, temperature and gas exchange. To investigate lymphocyte migration upon liver inflammation CXCR6.gfp knock-in mice were subjected to intravital liver imaging under baseline conditions and after acute and chronic liver damage induced by intraperitoneal injection(s) of carbon tetrachloride (CCl4). CXCR6 is a chemokine receptor expressed on lymphocytes, mainly on Natural Killer T (NKT)-, Natural Killer (NK)- and subsets of T lymphocytes such as CD4 T cells but also mucosal associated invariant (MAIT) T cells1. Following the migratory pattern and positioning of CXCR6.gfp+ immune cells allowed a

  16. High spectral resolution airborne short wave infrared hyperspectral imager

    NASA Astrophysics Data System (ADS)

    Wei, Liqing; Yuan, Liyin; Wang, Yueming; Zhuang, Xiaoqiong

    2016-05-01

    Short Wave InfraRed(SWIR) spectral imager is good at detecting difference between materials and penetrating fog and mist. High spectral resolution SWIR hyperspectral imager plays a key role in developing earth observing technology. Hyperspectral data cube can help band selections that is very important for multispectral imager design. Up to now, the spectral resolution of many SWIR hyperspectral imagers is about 10nm. A high sensitivity airborne SWIR hyperspectral imager with narrower spectral band will be presented. The system consists of TMA telescope, slit, spectrometer with planar blazed grating and high sensitivity MCT FPA. The spectral sampling interval is about 3nm. The IFOV is 0.5mrad. To eliminate the influence of the thermal background, a cold shield is designed in the dewar. The pixel number of spatial dimension is 640. Performance measurement in laboratory and image analysis for flight test will also be presented.

  17. Compressive spectral integral imaging using a microlens array

    NASA Astrophysics Data System (ADS)

    Feng, Weiyi; Rueda, Hoover; Fu, Chen; Qian, Chen; Arce, Gonzalo R.

    2016-05-01

    In this paper, a compressive spectral integral imaging system using a microlens array (MLA) is proposed. This system can sense the 4D spectro-volumetric information into a compressive 2D measurement image on the detector plane. In the reconstruction process, the 3D spatial information at different depths and the spectral responses of each spatial volume pixel can be obtained simultaneously. In the simulation, sensing of the 3D objects is carried out by optically recording elemental images (EIs) using a scanned pinhole camera. With the elemental images, a spectral data cube with different perspectives and depth information can be reconstructed using the TwIST algorithm in the multi-shot compressive spectral imaging framework. Then, the 3D spatial images with one dimensional spectral information at arbitrary depths are computed using the computational integral imaging method by inversely mapping the elemental images according to geometrical optics. The simulation results verify the feasibility of the proposed system. The 3D volume images and the spectral information of the volume pixels can be successfully reconstructed at the location of the 3D objects. The proposed system can capture both 3D volumetric images and spectral information in a video rate, which is valuable in biomedical imaging and chemical analysis.

  18. Spectral edge: gradient-preserving spectral mapping for image fusion.

    PubMed

    Connah, David; Drew, Mark S; Finlayson, Graham D

    2015-12-01

    This paper describes a novel approach to image fusion for color display. Our goal is to generate an output image whose gradient matches that of the input as closely as possible. We achieve this using a constrained contrast mapping paradigm in the gradient domain, where the structure tensor of a high-dimensional gradient representation is mapped exactly to that of a low-dimensional gradient field which is then reintegrated to form an output. Constraints on output colors are provided by an initial RGB rendering. Initially, we motivate our solution with a simple "ansatz" (educated guess) for projecting higher-D contrast onto color gradients, which we expand to a more rigorous theorem to incorporate color constraints. The solution to these constrained optimizations is closed-form, allowing for simple and hence fast and efficient algorithms. The approach can map any N-D image data to any M-D output and can be used in a variety of applications using the same basic algorithm. In this paper, we focus on the problem of mapping N-D inputs to 3D color outputs. We present results in five applications: hyperspectral remote sensing, fusion of color and near-infrared or clear-filter images, multilighting imaging, dark flash, and color visualization of magnetic resonance imaging diffusion-tensor imaging. PMID:26831392

  19. Hyperspectral retinal imaging with a spectrally tunable light source

    NASA Astrophysics Data System (ADS)

    Francis, Robert P.; Zuzak, Karel J.; Ufret-Vincenty, Rafael

    2011-03-01

    Hyperspectral retinal imaging can measure oxygenation and identify areas of ischemia in human patients, but the devices used by current researchers are inflexible in spatial and spectral resolution. We have developed a flexible research prototype consisting of a DLP®-based spectrally tunable light source coupled to a fundus camera to quickly explore the effects of spatial resolution, spectral resolution, and spectral range on hyperspectral imaging of the retina. The goal of this prototype is to (1) identify spectral and spatial regions of interest for early diagnosis of diseases such as glaucoma, age-related macular degeneration (AMD), and diabetic retinopathy (DR); and (2) define required specifications for commercial products. In this paper, we describe the challenges and advantages of using a spectrally tunable light source for hyperspectral retinal imaging, present clinical results of initial imaging sessions, and describe how this research can be leveraged into specifying a commercial product.

  20. Fast and simple spectral FLIM for biochemical and medical imaging.

    PubMed

    Popleteeva, Marina; Haas, Kalina T; Stoppa, David; Pancheri, Lucio; Gasparini, Leonardo; Kaminski, Clemens F; Cassidy, Liam D; Venkitaraman, Ashok R; Esposito, Alessandro

    2015-09-01

    Spectrally resolved fluorescence lifetime imaging microscopy (λFLIM) has powerful potential for biochemical and medical imaging applications. However, long acquisition times, low spectral resolution and complexity of λFLIM often narrow its use to specialized laboratories. Therefore, we demonstrate here a simple spectral FLIM based on a solid-state detector array providing in-pixel histrogramming and delivering faster acquisition, larger dynamic range, and higher spectral elements than state-of-the-art λFLIM. We successfully apply this novel microscopy system to biochemical and medical imaging demonstrating that solid-state detectors are a key strategic technology to enable complex assays in biomedical laboratories and the clinic.

  1. Spectral imaging of breast fibroadenoma using second-harmonic generation

    NASA Astrophysics Data System (ADS)

    Zheng, Liqin; Wang, Yuhua

    2014-09-01

    Fibroadenoma (FA), typically composed of stroma and epithelial cells, is a very common benign breast disease. Women with FA are associated with an increased risk of future breast cancer. The objective of this study was to demonstrate the potential of multiphoton laser scanning microscopy (MPLSM) for characterizing the morphology of collagen in the human breast fibroadenomas. In the study, high-contrast SHG images of human normal breast tissues and fibroadenoma tissues were obtained for comparison. The morphology of collagen was different between normal breast tissue and fibroadenoma. This study shows that MPLSM has the ability to distinguish fibroadenoma tissues from the normal breast tissues based on the noninvasive SHG imaging. With the advent of the clinical portability of miniature MPLSM, we believe that the technique has great potential to be used in vivo studies and for monitoring the treatment responses of fibroadenomas in clinical.

  2. Compact multi-spectral imaging system for dermatology and neurosurgery

    NASA Astrophysics Data System (ADS)

    Noordmans, Herke Jan; de Roode, Rowland; Verdaasdonk, Rudolf

    2007-03-01

    A compact multi-spectral imaging system is presented as diagnostic tool in dermatology and neurosurgery. Using an electronically tunable filter, a sensitive high resolution digital camera, 140 spectral images from 400 nm up to 720 nm are acquired in 40 s. Advanced image processing algorithms are used to enable interactive acquisition, viewing, image registration and image analysis. Experiments in the department of dermatology and neurosurgery show that multispectral imaging reveals much more detail than conventional medical photography or a surgical microscope, as images can be reprocessed to enhance the view on e.g. tumor boundaries. Using a hardware-based interactive registration algorithm, multi-spectral images can be aligned to correct for motion occurred during image acquisition or to compare acquisitions from different moments in time. The system shows to be a powerful diagnostics tool for medical imaging in the visual and near IR range.

  3. Real-time Ca ion wave imaging in living rat cardiac muscle cells by a confocal multiphoton microscope with a microlens-pinhole array scanner

    NASA Astrophysics Data System (ADS)

    Fujita, Katsumasa; Kaneko, Tomoyuki; Nakamura, Osamu; Oyamada, Masahito; Takamatsu, Tetsuro; Kawata, Satoshi

    2000-04-01

    A real-time confocal multiphoton fluorescence microscope was developed to observe Ca2+ dynamics in living rat- cardiac muscle cells. The real-time imaging was achieved by multifocus excitation of a specimen with a rotating microlens-array disk. A pinhole-array disk for confocal detection was introduced in the microscope to improve the spatial resolution and the contrast of fluorescence images. Ca2+ wave and Ca2+ transient in cultured rat- cardiac cells were successfully observed with the developed microscope.

  4. Passive Microwave Spectral Imaging of Amospheric Structure

    NASA Technical Reports Server (NTRS)

    Staelin, David H.; Rosenkranz, Philip W.

    1998-01-01

    The primary objective of this research was to improve the scientific foundation necessary to full realization of the meteorological potential of the NOAA Advanced Microwave Sounding Unit (AMSU) recently first launched on the NOAA-15 satellite in May, 1998. These advances were made in four main areas: (1) improvements, based on aircraft observations, in the atmospheric transmittance expressions used for interpreting AMSU and similar data; (2) development of neural network retrieval methods for cloud top altitude estimates of approximately 1-km accuracy under cirrus shields--the altitude is that of the larger ice particles aloft, which is related to precipitation rate; (3) analysis of early AMSU flight data with respect to its precipitation sensitivity and fine-scale thermal structure; and (4) improvements to the 54-GHz and 118-GHz MTS aircraft imaging spectrometer now operating on the NASA ER-2 aircraft. More specifically, the oxygen transmittance expressions near 118 GHz were in better agreement with aircraft data when the temperature dependence exponent of the 118.75-GHz linewidth was increased from the MPM92 value (Liebe et al., 1992) of 0.8 to 0.97+/-0.03. In contrast, the observations 52.5-55.8 GHz were consistent with the MPM92 model. Neural networks trained on comparisons of 118-GHz spectral data and coincident stereoscopic video images of convective cells observed from 20-km altitude yielded agreement in their peak altitudes within as little as 1.36 km rms, much of which is stereoscopic error. Imagery using these methods produced useful characterizations for Cyclone Oliver in 1993 and other storms (Schwartz et al., 1996; Spina et al., 1998). Similar neural network techniques yielded simulated rms errors in relative humidity retrievals of 6-14 percent over ocean and 6-15 percent over land at pressure levels from 1013 to 131 mbar (Cabrera-Mercader and Staelin, 1995).

  5. Maximum a posteriori estimation of spectral reflectance from color image and multipoint spectral measurements.

    PubMed

    Murakami, Yuri; Ietomi, Kunihiko; Yamaguchi, Masahiro; Ohyama, Nagaaki

    2007-10-01

    Accurate color image reproduction under arbitrary illumination can be realized if the spectral reflectance functions in a scene are obtained. Although multispectral imaging is one of the promising methods to obtain the reflectance of a scene, it is expected to reduce the number of color channels without significant loss of accuracy. This paper presents what we believe to be a new method for estimating spectral reflectance functions from color image and multipoint spectral measurements based on maximum a posteriori (MAP) estimation. Multipoint spectral measurements are utilized as auxiliary information to improve the accuracy of spectral reflectance estimated from image data. Through simulations, it is confirmed that the proposed method improves the estimation accuracy, particularly when a scene includes subjects that belong to various categories.

  6. SPECTRAL IMAGING OF GALAXY CLUSTERS WITH PLANCK

    SciTech Connect

    Bourdin, H.; Mazzotta, P.; Rasia, E.

    2015-12-20

    The Sunyaev–Zeldovich (SZ) effect is a promising tool for detecting the presence of hot gas out to the galaxy cluster peripheries. We developed a spectral imaging algorithm dedicated to the SZ observations of nearby galaxy clusters with Planck, with the aim of revealing gas density anisotropies related to the filamentary accretion of materials, or pressure discontinuities induced by the propagation of shock fronts. To optimize an unavoidable trade-off between angular resolution and precision of the SZ flux measurements, the algorithm performs a multi-scale analysis of the SZ maps as well as of other extended components, such as the cosmic microwave background (CMB) anisotropies and the Galactic thermal dust. The demixing of the SZ signal is tackled through kernel-weighted likelihood maximizations. The CMB anisotropies are further analyzed through a wavelet analysis, while the Galactic foregrounds and SZ maps are analyzed via a curvelet analysis that best preserves their anisotropic details. The algorithm performance has been tested against mock observations of galaxy clusters obtained by simulating the Planck High Frequency Instrument and by pointing at a few characteristic positions in the sky. These tests suggest that Planck should easily allow us to detect filaments in the cluster peripheries and detect large-scale shocks in colliding galaxy clusters that feature favorable geometry.

  7. Evaluating collagen morphology and pathological lipid deposition using multiphoton image statistics

    NASA Astrophysics Data System (ADS)

    Mostaço-Guidolin, Leila B.; Ko, Alex C.-T.; Wang, Fei; Tian, Hong; Hewko, Mark; Shiomi, Masashi; Major, Arkady; Sowa, Michael G.

    2014-02-01

    In this study we present a novel image analysis methodology to quantify and to classify morphological details in tissue collagen fibril organization and lipid deposition. Co-localized collagen (second harmonic, SHG) and lipid (coherent Raman, CARS) images of atherosclerotic artery walls were acquired by a supercontinuum-powered multi-modal nonlinear microscope. Textural features based on the first-order statistics (FOS) and gray level co-occurrence matrix (GLCM) parameters were extracted from the SHG and CARS images. Multi-group classifications based on support vector machine of SHG and CARS images were subsequently performed to investigate the potential of texture analysis in providing quantitative descriptors of structural and compositional changes during disease progression. Using a rabbit model, different collagen remodeling and lipid accumulation patterns in disease tissues can be successfully tracked using these image statistics, thus providing a robust foundation for classification. When the variation of the CARS image features were tracked against the age of the rabbit, it was noticed that older animals (advanced plaques) present a more complex necrotic core containing high-lipid extracellular structures with various shapes and distribution. With combined FOS and GLCM texture statistics, we achieved reliable classification of SHG and CARS images acquired from atherosclerotic arteries with >90% accuracy, sensitivity and specificity. The proposed image analysis methodology can also be applied in a wide range of applications to evaluate conditions involving collagen re-modeling and prominent lipid accumulation.

  8. Code aperture optimization for spectrally agile compressive imaging.

    PubMed

    Arguello, Henry; Arce, Gonzalo R

    2011-11-01

    Coded aperture snapshot spectral imaging (CASSI) provides a mechanism for capturing a 3D spectral cube with a single shot 2D measurement. In many applications selective spectral imaging is sought since relevant information often lies within a subset of spectral bands. Capturing and reconstructing all the spectral bands in the observed image cube, to then throw away a large portion of this data, is inefficient. To this end, this paper extends the concept of CASSI to a system admitting multiple shot measurements, which leads not only to higher quality of reconstruction but also to spectrally selective imaging when the sequence of code aperture patterns is optimized. The aperture code optimization problem is shown to be analogous to the optimization of a constrained multichannel filter bank. The optimal code apertures allow the decomposition of the CASSI measurement into several subsets, each having information from only a few selected spectral bands. The rich theory of compressive sensing is used to effectively reconstruct the spectral bands of interest from the measurements. A number of simulations are developed to illustrate the spectral imaging characteristics attained by optimal aperture codes.

  9. Spectrally resolved fluorescence lifetime imaging to investigate cell metabolism in malignant and nonmalignant oral mucosa cells

    NASA Astrophysics Data System (ADS)

    Rück, Angelika; Hauser, Carmen; Mosch, Simone; Kalinina, Sviatlana

    2014-09-01

    Fluorescence-guided diagnosis of tumor tissue is in many cases insufficient, because false positive results interfere with the outcome. Improvement through observation of cell metabolism might offer the solution, but needs a detailed understanding of the origin of autofluorescence. With respect to this, spectrally resolved multiphoton fluorescence lifetime imaging was investigated to analyze cell metabolism in metabolic phenotypes of malignant and nonmalignant oral mucosa cells. The time-resolved fluorescence characteristics of NADH were measured in cells of different origins. The fluorescence lifetime of bound and free NADH was calculated from biexponential fitting of the fluorescence intensity decay within different spectral regions. The mean lifetime was increased from nonmalignant oral mucosa cells to different squamous carcinoma cells, where the most aggressive cells showed the longest lifetime. In correlation with reports in the literature, the total amount of NADH seemed to be less for the carcinoma cells and the ratio of free/bound NADH was decreased from nonmalignant to squamous carcinoma cells. Moreover for squamous carcinoma cells a high concentration of bound NADH was found in cytoplasmic organelles (mainly mitochondria). This all together indicates that oxidative phosphorylation and a high redox potential play an important role in the energy metabolism of these cells.

  10. Spectral homogenization techniques for the hyperspectral image projector

    NASA Astrophysics Data System (ADS)

    Hillberry, Logan E.; Rice, Joseph P.

    2015-05-01

    In an effort to improve technology for performance testing and calibration of multispectral and hyperspectral imagers, the National Institute of Standards and Technology (NIST) has been developing a Hyperspectral Image Projector (HIP) capable of projecting dynamic scenes than include distinct, programmable spectra in each of its 1024x768 spatial pixels. The HIP is comprised of a spectral engine, which is a light source capable generating the spectra in the scene, coupled to a spatial engine, capable of projecting the spectra into the correct locations of the scene. In the prototype HIP, the light exiting the Visible-Near-Infrared (VNIR) / Short-Wavelength Infrared (SWIR) spectral engine is spectrally dispersed and needs to be spectrally homogenized before it enters the spatial engine. In this paper we describe the results from a study of several different techniques for performing this spectral homogenization. These techniques include an integrating sphere, a liquid light guide, a randomized fiber bundle, and an engineered diffuser, in various combinations. The spectral uniformity of projected HIP scenes is measured and analyzed using the spectral angle mapper (SAM) algorithm over the VNIR spectral range. The SAM provides a way to analyze the spectral uniformity independently from the radiometric uniformity. The goal of the homogenizer is a spectrally uniform and bright projected image. An integrating sphere provides the most spectrally uniform image, but at a great loss of light compared with the other methods. The randomized fiber bundle generally outperforms the liquid light guide in both spectral homogenization and brightness. Using an engineered diffuser with the randomized fiber bundle increases the spectral uniformity by a factor of five, with a decrease in brightness by a factor of five, compared with the randomized fiber bundle alone. The combination of an engineered diffuser with a randomized fiber bundle provides comparable spectral uniformity to the

  11. SAR image change detection using watershed and spectral clustering

    NASA Astrophysics Data System (ADS)

    Niu, Ruican; Jiao, L. C.; Wang, Guiting; Feng, Jie

    2011-12-01

    A new method of change detection in SAR images based on spectral clustering is presented in this paper. Spectral clustering is employed to extract change information from a pair images acquired on the same geographical area at different time. Watershed transform is applied to initially segment the big image into non-overlapped local regions, leading to reduce the complexity. Experiments results and system analysis confirm the effectiveness of the proposed algorithm.

  12. Motion quantification during multi-photon functional imaging in behaving animals

    PubMed Central

    Kong, Lingjie; Little, Justin P.; Cui, Meng

    2016-01-01

    Functional imaging in behaving animals is essential to understanding brain function. However, artifacts resulting from animal motion, including locomotion, can severely corrupt functional measurements. To dampen tissue motion, we designed a new optical window with minimal optical aberrations. Using the newly developed high-speed continuous volumetric imaging system based on an optical phase-locked ultrasound lens, we quantified motion of the cerebral cortex and hippocampal surface during two-photon functional imaging in behaving mice. We find that the out-of-plane motion is generally greater than the axial dimension of the point-spread-function during mouse locomotion, which indicates that high-speed continuous volumetric imaging is necessary to minimize motion artifacts.

  13. Motion quantification during multi-photon functional imaging in behaving animals

    PubMed Central

    Kong, Lingjie; Little, Justin P.; Cui, Meng

    2016-01-01

    Functional imaging in behaving animals is essential to understanding brain function. However, artifacts resulting from animal motion, including locomotion, can severely corrupt functional measurements. To dampen tissue motion, we designed a new optical window with minimal optical aberrations. Using the newly developed high-speed continuous volumetric imaging system based on an optical phase-locked ultrasound lens, we quantified motion of the cerebral cortex and hippocampal surface during two-photon functional imaging in behaving mice. We find that the out-of-plane motion is generally greater than the axial dimension of the point-spread-function during mouse locomotion, which indicates that high-speed continuous volumetric imaging is necessary to minimize motion artifacts. PMID:27699129

  14. Hyperspectral image classification by a variable interval spectral average and spectral curve matching combined algorithm

    NASA Astrophysics Data System (ADS)

    Senthil Kumar, A.; Keerthi, V.; Manjunath, A. S.; Werff, Harald van der; Meer, Freek van der

    2010-08-01

    Classification of hyperspectral images has been receiving considerable attention with many new applications reported from commercial and military sectors. Hyperspectral images are composed of a large number of spectral channels, and have the potential to deliver a great deal of information about a remotely sensed scene. However, in addition to high dimensionality, hyperspectral image classification is compounded with a coarse ground pixel size of the sensor for want of adequate sensor signal to noise ratio within a fine spectral passband. This makes multiple ground features jointly occupying a single pixel. Spectral mixture analysis typically begins with pixel classification with spectral matching techniques, followed by the use of spectral unmixing algorithms for estimating endmembers abundance values in the pixel. The spectral matching techniques are analogous to supervised pattern recognition approaches, and try to estimate some similarity between spectral signatures of the pixel and reference target. In this paper, we propose a spectral matching approach by combining two schemes—variable interval spectral average (VISA) method and spectral curve matching (SCM) method. The VISA method helps to detect transient spectral features at different scales of spectral windows, while the SCM method finds a match between these features of the pixel and one of library spectra by least square fitting. Here we also compare the performance of the combined algorithm with other spectral matching techniques using a simulated and the AVIRIS hyperspectral data sets. Our results indicate that the proposed combination technique exhibits a stronger performance over the other methods in the classification of both the pure and mixed class pixels simultaneously.

  15. Spectral ladar: towards active 3D multispectral imaging

    NASA Astrophysics Data System (ADS)

    Powers, Michael A.; Davis, Christopher C.

    2010-04-01

    In this paper we present our Spectral LADAR concept, an augmented implementation of traditional LADAR. This sensor uses a polychromatic source to obtain range-resolved 3D spectral images which are used to identify objects based on combined spatial and spectral features, resolving positions in three dimensions and up to hundreds of meters in distance. We report on a proof-of-concept Spectral LADAR demonstrator that generates spectral point clouds from static scenes. The demonstrator transmits nanosecond supercontinuum pulses generated in a photonic crystal fiber. Currently we use a rapidly tuned receiver with a high-speed InGaAs APD for 25 spectral bands with the future expectation of implementing a linear APD array spectrograph. Each spectral band is independently range resolved with multiple return pulse recognition. This is a critical feature, enabling simultaneous spectral and spatial unmixing of partially obscured objects when not achievable using image fusion of monochromatic LADAR and passive spectral imagers. This enables higher identification confidence in highly cluttered environments such as forested or urban areas (e.g. vehicles behind camouflage or foliage). These environments present challenges for situational awareness and robotic perception which can benefit from the unique attributes of Spectral LADAR. Results from this demonstrator unit are presented for scenes typical of military operations and characterize the operation of the device. The results are discussed here in the context of autonomous vehicle navigation and target recognition.

  16. Semiconductor Laser Multi-Spectral Sensing and Imaging

    PubMed Central

    Le, Han Q.; Wang, Yang

    2010-01-01

    Multi-spectral laser imaging is a technique that can offer a combination of the laser capability of accurate spectral sensing with the desirable features of passive multispectral imaging. The technique can be used for detection, discrimination, and identification of objects by their spectral signature. This article describes and reviews the development and evaluation of semiconductor multi-spectral laser imaging systems. Although the method is certainly not specific to any laser technology, the use of semiconductor lasers is significant with respect to practicality and affordability. More relevantly, semiconductor lasers have their own characteristics; they offer excellent wavelength diversity but usually with modest power. Thus, system design and engineering issues are analyzed for approaches and trade-offs that can make the best use of semiconductor laser capabilities in multispectral imaging. A few systems were developed and the technique was tested and evaluated on a variety of natural and man-made objects. It was shown capable of high spectral resolution imaging which, unlike non-imaging point sensing, allows detecting and discriminating objects of interest even without a priori spectroscopic knowledge of the targets. Examples include material and chemical discrimination. It was also shown capable of dealing with the complexity of interpreting diffuse scattered spectral images and produced results that could otherwise be ambiguous with conventional imaging. Examples with glucose and spectral imaging of drug pills were discussed. Lastly, the technique was shown with conventional laser spectroscopy such as wavelength modulation spectroscopy to image a gas (CO). These results suggest the versatility and power of multi-spectral laser imaging, which can be practical with the use of semiconductor lasers. PMID:22315555

  17. Real-time histology in liver disease using multiphoton microscopy with fluorescence lifetime imaging

    PubMed Central

    Wang, Haolu; Liang, Xiaowen; Mohammed, Yousuf H.; Thomas, James A.; Bridle, Kim R.; Thorling, Camilla A.; Grice, Jeffrey E.; Xu, Zhi Ping; Liu, Xin; Crawford, Darrell H. G.; Roberts, Michael S.

    2015-01-01

    Conventional histology with light microscopy is essential in the diagnosis of most liver diseases. Recently, a concept of real-time histology with optical biopsy has been advocated. In this study, live mice livers (normal, with fibrosis, steatosis, hepatocellular carcinoma and ischemia-reperfusion injury) were imaged by MPM-FLIM for stain-free real-time histology. The acquired MPM-FLIM images were compared with conventional histological images. MPM-FLIM imaged subsurface cellular and subcellular histopathological hallmarks of live liver in mice models at high resolution. Additional information such as distribution of stellate cell associated autofluorescence and fluorescence lifetime changes was also gathered by MPM-FLIM simultaneously, which cannot be obtained from conventional histology. MPM-FLIM could simultaneously image and quantify the cellular morphology and microenvironment of live livers without conventional biopsy or fluorescent dyes. We anticipate that in the near future MPM-FLIM will be evaluated from bench to bedside, leading to real-time histology and dynamic monitoring of human liver diseases. PMID:25798303

  18. Multiplexed Spectral Imaging of 120 Different Fluorescent Labels

    PubMed Central

    Valm, Alex M.; Oldenbourg, Rudolf; Borisy, Gary G.

    2016-01-01

    The number of fluorescent labels that can unambiguously be distinguished in a single image when acquired through band pass filters is severely limited by the spectral overlap of available fluorophores. The recent development of spectral microscopy and the application of linear unmixing algorithms to spectrally recorded image data have allowed simultaneous imaging of fluorophores with highly overlapping spectra. However, the number of distinguishable fluorophores is still limited by the unavoidable decrease in signal to noise ratio when fluorescence signals are fractionated over multiple wavelength bins. Here we present a spectral image analysis algorithm to greatly expand the number of distinguishable objects labeled with binary combinations of fluorophores. Our algorithm utilizes a priori knowledge about labeled specimens and imposes a binary label constraint on the unmixing solution. We have applied our labeling and analysis strategy to identify microbes labeled by fluorescence in situ hybridization and here demonstrate the ability to distinguish 120 differently labeled microbes in a single image. PMID:27391327

  19. A Wide Spectral Range Reflectance and Luminescence Imaging System

    PubMed Central

    Hirvonen, Tapani; Penttinen, Niko; Hauta–Kasari, Markku; Sorjonen, Mika; Peiponen, Kai–Erik

    2013-01-01

    In this study, we introduce a wide spectral range (200–2500 nm) imaging system with a 250 μm minimum spatial resolution, which can be freely modified for a wide range of resolutions and measurement geometries. The system has been tested for reflectance and luminescence measurements, but can also be customized for transmittance measurements. This study includes the performance results of the developed system, as well as examples of spectral images. Discussion of the system relates it to existing systems and methods. The wide range spectral imaging system that has been developed is however highly customizable and has great potential in many practical applications. PMID:24233075

  20. A novel clinical multimodal multiphoton tomograph for AF, SHG, CARS imaging, and FLIM

    NASA Astrophysics Data System (ADS)

    Weinigel, Martin; Breunig, Hans Georg; König, Karsten

    2014-02-01

    We report on a flexible nonlinear medical tomograph with multiple miniaturized detectors for simultaneous acquisition of two-photon autofluorescence (AF), second harmonic generation (SHG) and coherent anti-Stokes Raman scattering (CARS) images. The simultaneous visualization of the distribution of endogenous fluorophores NAD(P)H, melanin and elastin, SHG-active collagen and as well as non-fluorescent lipids within human skin in vivo is possible. Furthermore, fluorescence lifetime images (FLIM) can be generated using time-correlated single photon counting.

  1. Multiphotonic Confocal Microscopy 3D imaging: Application to mantle sulfides in sub-arc environment (Avacha Volcano, Kamchatka)

    NASA Astrophysics Data System (ADS)

    Antoine, Bénard; Luc-Serge, Doucet; Sabine, Palle; Dmitri A., Ionov

    2010-05-01

    Petrogenetic relations in igneous rocks are usually studied in natural samples using classical optical microscopy and subsequent geochemical data acquisition. Multiphotonic Laser Scanning Confocal Microscopy (MLSCM) can be a powerful tool to section geological materials optically with sub-micrometric resolution and then generate a three-dimensional (3D) reconstruction (ca. 106 μm3 stack). MLSCM is used here to investigate textural relations of Monosulfide Solid Solution (MSS) with silicate phases in fresh spinel harzburgite xenoliths from the andesitic Avacha volcano (Kamchatka, Russia). The xenoliths contain MSS disseminated in olivine and orthopyroxene (opx) neoblasts as well as MSS-rich quenched magmatic opx veins [1]. First, Reflection Mode (RM) was tested on vein sulfides in resin-impregnated thick (120 μm) polished rock sections. Then we used a combination of Differential Interference Contrast (DIC) with a transmitted light detector, two photons-excited fluorescence (2PEF) and Second Harmonic Generation (SHG). Sequential imaging feature of the Leica TCS-SP2 software was applied. The excitation laser used for 2PEF was a COHERENT MIRA 900 with a 76Hz repetition rate and 800nm wavelength. Image stacks were analysed using ImageJ software [2]. The aim of the tests was to try to discriminate sulfides in silicate matrix as a tool for a better assessment of equilibrium conditions between the two phases. Preliminary results show that Fe-Ni rich MSS from vein and host rock have a strong auto-fluorescence in the Near UV-VIS domain (392-715 nm) whereas silicate matrix is only revealed through DIC. SHG is obtained only from dense nanocentrosymmetrical structures such as embedded medium (organic matter like glue and resin). The three images were recorded sequentially enabling efficient discrimination between the different components of the rock slices. RM permits reconstruction of the complete 3D structure of the rock slice. High resolution (ca. 0.2 μm along X-Y axis vs

  2. [Research on Spectral Polarization Imaging System Based on Static Modulation].

    PubMed

    Zhao, Hai-bo; Li, Huan; Lin, Xu-ling; Wang, Zheng

    2015-04-01

    The main disadvantages of traditional spectral polarization imaging system are: complex structure, with moving parts, low throughput. A novel method of spectral polarization imaging system is discussed, which is based on static polarization intensity modulation combined with Savart polariscope interference imaging. The imaging system can obtain real-time information of spectral and four Stokes polarization messages. Compared with the conventional methods, the advantages of the imaging system are compactness, low mass and no moving parts, no electrical control, no slit and big throughput. The system structure and the basic theory are introduced. The experimental system is established in the laboratory. The experimental system consists of reimaging optics, polarization intensity module, interference imaging module, and CCD data collecting and processing module. The spectral range is visible and near-infrared (480-950 nm). The white board and the plane toy are imaged by using the experimental system. The ability of obtaining spectral polarization imaging information is verified. The calibration system of static polarization modulation is set up. The statistical error of polarization degree detection is less than 5%. The validity and feasibility of the basic principle is proved by the experimental result. The spectral polarization data captured by the system can be applied to object identification, object classification and remote sensing detection. PMID:26197616

  3. [Spectral radiometric calibration research of Quick Bird digital image].

    PubMed

    Zhang, Guo-Kun; Chen, Chun; Xing, Fu; Zhang, Hong-Yan; Zhao, Yun-Sheng

    2008-03-01

    The present article uses the basic operation of the digital remote image radiometric calibration of the Quickbird with high distinguishing rate, including the physical attribute and the mathematical basement of digital images, the annotation as well as the format of image data. The study makes use of information of spectral radiance from the ground-atmosphere system, which is recorded by the digital remote image of Quick Bird in Honghe area. This dissertation offered the calculation means of radiometric calibration, and changed the pixel digital number into band-integrated radiance. Then, the spectral radiance was calculated. After the radiometric calibration, the Quick Bird image showed the quantitative information of spectral feature from various ground items. Only through the calibration can the Quick Bird image be quantitatively compared and analyzed with other remote sensor images. Thus, the inversion image has the value of application. The significance consists in offering important basic condition for the image amalgamation and better disposal of the special inforation pick-up. This effort also offered spectral information of the ground items for the inversion of the remote image. Therefore, the authors can combine the research of the spectral character of ground items with the establishment of the remote application model in order to quantitatively analyze the ground items.

  4. Multiphoton tomography for tissue engineering

    NASA Astrophysics Data System (ADS)

    König, Karsten

    2008-02-01

    Femtosecond laser multiphoton tomography has been employed in the field of tissue engineering to perform 3D high-resolution imaging of the extracellular matrix proteins elastin and collagen as well as of living cells without any fixation, slicing, and staining. Near infrared 80 MHz picojoule femtosecond laser pulses are able to excite the endogenous fluorophores NAD(P)H, flavoproteins, melanin, and elastin via a non-resonant two-photon excitation process. In addition, collagen can be imaged by second harmonic generation. Using a two-PMT detection system, the ratio of elastin to collagen was determined during optical sectioning. A high submicron spatial resolution and 50 picosecond temporal resolution was achieved using galvoscan mirrors and piezodriven focusing optics as well as a time-correlated single photon counting module with a fast microchannel plate detector and fast photomultipliers. Multiphoton tomography has been used to optimize the tissue engineering of heart valves and vessels in bioincubators as well as to characterize artificial skin. Stem cell characterization and manipulation are of major interest for the field of tissue engineering. Using the novel sub-20 femtosecond multiphoton nanoprocessing laser microscope FemtOgene, the differentiation of human stem cells within spheroids has been in vivo monitored with submicron resolution. In addition, the efficient targeted transfection has been demonstrated. Clinical studies on the interaction of tissue-engineered products with the natural tissue environment can be performed with in vivo multiphoton tomograph DermaInspect.

  5. USE OF MULTIPHOTON LASER SCANNING MICROSCOPY TO IMAGE BENZO[A]PYRENE AND METABOLITES IN FISH EARLY LIFE STAGES

    EPA Science Inventory

    Multiphoton laser scanning micrsocopy holds promise as a tool to study the tissue distribution of environmental chemical contaminants during fish early life stage development. One such chemical for which this is possible is benzo[a]pyrene (BaP), a polyaromatic hydrocarbon that a...

  6. USE OF MULTIPHOTON LASER SCANNING MICROSCOPY TO IMAGE BENZO[A]PYRENE AND METABOLITES IN FISH EGGS

    EPA Science Inventory

    Multiphoton laser scanning microscopy (MPLSM) is a promising tool to study the tissue distribution of environmental chemical contaminants during fish early life stages. One such chemical for which this is possible is benzo[a]pyrene (BaP), a polycyclic aromatic hydrocarbon that a...

  7. Infrared adaptive spectral imagers for direct detection of spectral signatures and hyperspectral imagery

    NASA Astrophysics Data System (ADS)

    Goldstein, Neil; Fox, Marsha; Adler-Golden, Steven; Gregor, Brian

    2013-03-01

    Field test results are presented for a prototype long-wave adaptive imager that provides both hyperspectral imagery and contrast imagery based on the direct application of hyperspectral detection algorithms in hardware. Programmable spatial light modulators are used to provide both spectral and spatial resolution using a single element detector. Programmable spectral and spatial detection filters can be used to superimpose any possible analog spectral detection filter on the image. In this work, we demonstrate three modes of operation, including hyperspectral imagery, and one and two-dimensional imagery using a generalized matched filter for detection of a specific target gas within the scene.

  8. An integrated coherent anti-Stokes Raman scattering and multiphoton imaging technique for liver disease diagnosis

    NASA Astrophysics Data System (ADS)

    Lin, Jian; Lu, Fake; Zheng, Wei; Yu, Hanry; Sheppard, Colin; Huang, Zhiwei

    2012-03-01

    Liver steatosis and fibrosis are two prevalence liver diseases and may eventually develop into hepatocellular carcinoma (HCC) Due to their prevalence and severity, much work has been done to develop efficient diagnostic methods and therapies. Nonlinear optical microscopy has high sensitivity and chemical specificity for major biochemical compounds, making it a powerful tool for tissue imaging without staining. In this study, three nonlinear microscopy imaging modalities are applied to the study of liver diseases in a bile duct ligation rat modal. CARS shows the distributions of fats or lipids quantitatively across the tissue; SHG visualizes the collagens; and TPEF reveals the morphology of hepatic cells. The results clearly show the development of liver steatosis and fibrosis with time, and the hepatic fat and collagen fibrils are quantified. This study demonstrates the ability of multimodal nonlinear optical microscopy for liver disease diagnosis, and may provide new insights into the understanding of the mechanisms of steatosis/fibrosis transformations at the cellular and molecular levels.

  9. A novel method for observing proteins in vivo using a small fluorescent label and multiphoton imaging.

    PubMed

    Botchway, Stanley W; Barba, Ignasi; Jordan, Randolf; Harmston, Rebecca; Haggie, Peter M; Williams, Simon-Peter; Fulton, Alexandra M; Parker, Anthony W; Brindle, Kevin M

    2005-09-15

    A novel method for the fluorescence detection of proteins in cells is described in the present study. Proteins are labelled by the selective biosynthetic incorporation of 5-hydroxytryptophan and the label is detected via selective two-photon excitation of the hydroxyindole and detection of its fluorescence emission at 340 nm. The method is demonstrated in this paper with images of a labelled protein in yeast cells.

  10. In vivo multiphoton imaging reveals gradual growth of newborn amyloid plaques over weeks.

    PubMed

    Burgold, Steffen; Bittner, Tobias; Dorostkar, Mario M; Kieser, Daniel; Fuhrmann, Martin; Mitteregger, Gerda; Kretzschmar, Hans; Schmidt, Boris; Herms, Jochen

    2011-03-01

    The kinetics of amyloid plaque formation and growth as one of the characteristic hallmarks of Alzheimer's disease (AD) are fundamental issues in AD research. Especially the question how fast amyloid plaques grow to their final size after they are born remains controversial. By long-term two-photon in vivo imaging we monitored individual methoxy-X04-stained amyloid plaques over 6 weeks in 12 and 18 months old Tg2576 mice. We found that in 12 months old mice, newly appearing amyloid plaques were initially small in volume and subsequently grew over time. The growth rate of plaques was inversely proportional to their volume; thus amyloid plaques that were already present at the first imaging time point grew over time but slower compared to new plaques. Additionally, we analyzed 18 months old Tg2576 mice in which we neither found newly appearing plaques nor a significant growth of pre-existing plaques over 6 weeks of imaging. In conclusion, newly appearing amyloid plaques are initially small in size but grow over time until plaque growth can not be detected anymore in aged mice. These results suggest that drugs that target plaque formation should be most effective early in the disease, when plaques are growing.

  11. Multiphoton fluorescence lifetime imaging of metabolic status in mesenchymal stem cell during adipogenic differentiation

    NASA Astrophysics Data System (ADS)

    Meleshina, A. V.; Dudenkova, V. V.; Shirmanova, M. V.; Bystrova, A. S.; Zagaynova, E. V.

    2016-03-01

    Non-invasive imaging of cell metabolism is a valuable approach to assess the efficacy of stem cell therapy and understand the tissue development. In this study we analyzed metabolic trajectory of the mesenchymal stem cells (MCSs) during differentiation into adipocytes by measuring fluorescence lifetimes of free and bound forms of the reduced nicotinamide adenine dinucleotide (NAD(P)H) and flavine adenine dinucleotide (FAD). Undifferentiated MSCs and MSCs on the 5, 12, 19, 26 days of differentiation were imaged on a Zeiss 710 microscope with fluorescence lifetime imaging (FLIM) system B&H (Germany). Fluorescence of NAD(P)H and FAD was excited at 750 nm and 900 nm, respectively, by a femtosecond Ti:sapphire laser and detected in a range 455-500 nm and 500-550 nm, correspondingly. We observed the changes in the NAD(P)H and FAD fluorescence lifetimes and their relative contributions in the differentiated adipocytes compare to undifferentiated MSCs. Increase of fluorescence lifetimes of the free and bound forms of NAD(P)H and the contribution of protein-bound NAD(P)H was registered, that can be associated with a metabolic switch from glycolysis to oxidative phosphorylation and/or synthesis of lipids in adipogenically differentiated MSCs. We also found that the contribution of protein-bound FAD decreased during differentiation. After carrying out appropriate biochemical measurements, the observed changes in cellular metabolism can potentially serve to monitor stem cell differentiation by FLIM.

  12. The application of fluorescent quantum dots to confocal, multiphoton, and electron microscopic imaging.

    PubMed

    Deerinck, Thomas J

    2008-01-01

    Fluorescent quantum dots are emerging as an important tool for imaging cells and tissues, and their unique optical and physical properties have captured the attention of the research community. The most common types of commercially available quantum dots consist of a nanocrystalline semiconductor core composed of cadmium selenide with a zinc sulfide capping layer and an outer polymer layer to facilitate conjugation to targeting biomolecules such as immunoglobulins. They exhibit high fluorescent quantum yields and have large absorption cross-sections, possess excellent photostability, and can be synthesized so that their narrow-band fluorescence emission can occur in a wide spectrum of colors. These properties make them excellent candidates for serving as multiplexing molecular beacons using a variety of imaging modalities including highly correlated microscopies. Whereas much attention has been focused on quantum-dot applications for live-cell imaging, we have sought to characterize and exploit their utility for enabling simultaneous multiprotein immunolabeling in fixed cells and tissues. Considerations for their application to immunolabeling for correlated light and electron microscopic analysis are discussed. PMID:18337229

  13. Spectral reflectance and digital image relations among five aquatic weeds

    Technology Transfer Automated Retrieval System (TEKTRAN)

    This study reports on the use of an artificial quartz halogen lighting source to facilitate the acquisition of spectral light reflectance measurements and digital imaging of invasive aquatic weeds. Spectral leaf or leaf/stem reflectance measurements were made on five aquatic weeds: Eurasian watermil...

  14. Colored coded-apertures for spectral image unmixing

    NASA Astrophysics Data System (ADS)

    Vargas, Hector M.; Arguello Fuentes, Henry

    2015-10-01

    Hyperspectral remote sensing technology provides detailed spectral information from every pixel in an image. Due to the low spatial resolution of hyperspectral image sensors, and the presence of multiple materials in a scene, each pixel can contain more than one spectral signature. Therefore, endmember extraction is used to determine the pure spectral signature of the mixed materials and its corresponding abundance map in a remotely sensed hyperspectral scene. Advanced endmember extraction algorithms have been proposed to solve this linear problem called spectral unmixing. However, such techniques require the acquisition of the complete hyperspectral data cube to perform the unmixing procedure. Researchers show that using colored coded-apertures improve the quality of reconstruction in compressive spectral imaging (CSI) systems under compressive sensing theory (CS). This work aims at developing a compressive supervised spectral unmixing scheme to estimate the endmembers and the abundance map from compressive measurements. The compressive measurements are acquired by using colored coded-apertures in a compressive spectral imaging system. Then a numerical procedure estimates the sparse vector representation in a 3D dictionary by solving a constrained sparse optimization problem. The 3D dictionary is formed by a 2-D wavelet basis and a known endmembers spectral library, where the Wavelet basis is used to exploit the spatial information. The colored coded-apertures are designed such that the sensing matrix satisfies the restricted isometry property with high probability. Simulations show that the proposed scheme attains comparable results to the full data cube unmixing technique, but using fewer measurements.

  15. New spectral imaging techniques for blood oximetry in the retina

    NASA Astrophysics Data System (ADS)

    Alabboud, Ied; Muyo, Gonzalo; Gorman, Alistair; Mordant, David; McNaught, Andrew; Petres, Clement; Petillot, Yvan R.; Harvey, Andrew R.

    2007-07-01

    Hyperspectral imaging of the retina presents a unique opportunity for direct and quantitative mapping of retinal biochemistry - particularly of the vasculature where blood oximetry is enabled by the strong variation of absorption spectra with oxygenation. This is particularly pertinent both to research and to clinical investigation and diagnosis of retinal diseases such as diabetes, glaucoma and age-related macular degeneration. The optimal exploitation of hyperspectral imaging however, presents a set of challenging problems, including; the poorly characterised and controlled optical environment of structures within the retina to be imaged; the erratic motion of the eye ball; and the compounding effects of the optical sensitivity of the retina and the low numerical aperture of the eye. We have developed two spectral imaging techniques to address these issues. We describe first a system in which a liquid crystal tuneable filter is integrated into the illumination system of a conventional fundus camera to enable time-sequential, random access recording of narrow-band spectral images. Image processing techniques are described to eradicate the artefacts that may be introduced by time-sequential imaging. In addition we describe a unique snapshot spectral imaging technique dubbed IRIS that employs polarising interferometry and Wollaston prism beam splitters to simultaneously replicate and spectrally filter images of the retina into multiple spectral bands onto a single detector array. Results of early clinical trials acquired with these two techniques together with a physical model which enables oximetry map are reported.

  16. Design of visible/infrared double-band spectral imager

    NASA Astrophysics Data System (ADS)

    Tianjin, Tang; Zhuo, Zhang; Bao-hua, Wang

    2016-01-01

    OFFNER hyperspectral imager using convex grating as spectral splitting component has many merits including large relative aperture, no smile, small key stone, compact construction and easier assembly and etc, has widely used in many occasions of various fields. Design of double-band hyperspectral imager using OFFNER structure is presented in this paper. SHAFER fore telescope system and convex grating imaging are adopted, groove etching density is different in different region on the same substrate for different spectral band, and different diffractive order is used respectively to split the two spectral bands. The optical system achieves good image quality in wide spectral range, compact construction and small volume. And all the reflective curved face are sphere, moderate tolerances are good for the processing and alignment.

  17. Digital staining for histopathology multispectral images by the combined application of spectral enhancement and spectral transformation.

    PubMed

    Bautista, Pinky A; Yagi, Yukako

    2011-01-01

    In this paper we introduced a digital staining method for histopathology images captured with an n-band multispectral camera. The method consisted of two major processes: enhancement of the original spectral transmittance and the transformation of the enhanced transmittance to its target spectral configuration. Enhancement is accomplished by shifting the original transmittance with the scaled difference between the original transmittance and the transmittance estimated with m dominant principal component (PC) vectors;the m-PC vectors were determined from the transmittance samples of the background image. Transformation of the enhanced transmittance to the target spectral configuration was done using an nxn transformation matrix, which was derived by applying a least square method to the enhanced and target spectral training data samples of the different tissue components. Experimental results on the digital conversion of a hematoxylin and eosin (H&E) stained multispectral image to its Masson's trichrome stained (MT) equivalent shows the viability of the method.

  18. Retinex Preprocessing for Improved Multi-Spectral Image Classification

    NASA Technical Reports Server (NTRS)

    Thompson, B.; Rahman, Z.; Park, S.

    2000-01-01

    The goal of multi-image classification is to identify and label "similar regions" within a scene. The ability to correctly classify a remotely sensed multi-image of a scene is affected by the ability of the classification process to adequately compensate for the effects of atmospheric variations and sensor anomalies. Better classification may be obtained if the multi-image is preprocessed before classification, so as to reduce the adverse effects of image formation. In this paper, we discuss the overall impact on multi-spectral image classification when the retinex image enhancement algorithm is used to preprocess multi-spectral images. The retinex is a multi-purpose image enhancement algorithm that performs dynamic range compression, reduces the dependence on lighting conditions, and generally enhances apparent spatial resolution. The retinex has been successfully applied to the enhancement of many different types of grayscale and color images. We show in this paper that retinex preprocessing improves the spatial structure of multi-spectral images and thus provides better within-class variations than would otherwise be obtained without the preprocessing. For a series of multi-spectral images obtained with diffuse and direct lighting, we show that without retinex preprocessing the class spectral signatures vary substantially with the lighting conditions. Whereas multi-dimensional clustering without preprocessing produced one-class homogeneous regions, the classification on the preprocessed images produced multi-class non-homogeneous regions. This lack of homogeneity is explained by the interaction between different agronomic treatments applied to the regions: the preprocessed images are closer to ground truth. The principle advantage that the retinex offers is that for different lighting conditions classifications derived from the retinex preprocessed images look remarkably "similar", and thus more consistent, whereas classifications derived from the original

  19. A parametric estimation approach to instantaneous spectral imaging.

    PubMed

    Oktem, Figen S; Kamalabadi, Farzad; Davila, Joseph M

    2014-12-01

    Spectral imaging, the simultaneous imaging and spectroscopy of a radiating scene, is a fundamental diagnostic technique in the physical sciences with widespread application. Due to the intrinsic limitation of two-dimensional (2D) detectors in capturing inherently three-dimensional (3D) data, spectral imaging techniques conventionally rely on a spatial or spectral scanning process, which renders them unsuitable for dynamic scenes. In this paper, we present a nonscanning (instantaneous) spectral imaging technique that estimates the physical parameters of interest by combining measurements with a parametric model and solving the resultant inverse problem computationally. The associated inverse problem, which can be viewed as a multiframe semiblind deblurring problem (with shift-variant blur), is formulated as a maximum a posteriori (MAP) estimation problem since in many such experiments prior statistical knowledge of the physical parameters can be well estimated. Subsequently, an efficient dynamic programming algorithm is developed to find the global optimum of the nonconvex MAP problem. Finally, the algorithm and the effectiveness of the spectral imaging technique are illustrated for an application in solar spectral imaging. Numerical simulation results indicate that the physical parameters can be estimated with the same order of accuracy as state-of-the-art slit spectroscopy but with the added benefit of an instantaneous, 2D field-of-view. This technique will be particularly useful for studying the spectra of dynamic scenes encountered in space remote sensing. PMID:25347878

  20. Snapshot spectral and polarimetric imaging; target identification with multispectral video

    NASA Astrophysics Data System (ADS)

    Bartlett, Brent D.; Rodriguez, Mikel D.

    2013-05-01

    As the number of pixels continue to grow in consumer and scientific imaging devices, it has become feasible to collect the incident light field. In this paper, an imaging device developed around light field imaging is used to collect multispectral and polarimetric imagery in a snapshot fashion. The sensor is described and a video data set is shown highlighting the advantage of snapshot spectral imaging. Several novel computer vision approaches are applied to the video cubes to perform scene characterization and target identification. It is shown how the addition of spectral and polarimetric data to the video stream allows for multi-target identification and tracking not possible with traditional RGB video collection.

  1. Stochastic scanning multiphoton multifocal microscopy.

    PubMed

    Jureller, Justin E; Kim, Hee Y; Scherer, Norbert F

    2006-04-17

    Multiparticle tracking with scanning confocal and multiphoton fluorescence imaging is increasingly important for elucidating biological function, as in the transport of intracellular cargo-carrying vesicles. We demonstrate a simple rapid-sampling stochastic scanning multifocal multiphoton microscopy (SS-MMM) fluorescence imaging technique that enables multiparticle tracking without specialized hardware at rates 1,000 times greater than conventional single point raster scanning. Stochastic scanning of a diffractive optic generated 10x10 hexagonal array of foci with a white noise driven galvanometer yields a scan pattern that is random yet space-filling. SS-MMM creates a more uniformly sampled image with fewer spatio-temporal artifacts than obtained by conventional or multibeam raster scanning. SS-MMM is verified by simulation and experimentally demonstrated by tracking microsphere diffusion in solution. PMID:19516485

  2. Hyperspectral imaging and spectral unmixing of stained tissue sections using a spectrally programmable light engine

    NASA Astrophysics Data System (ADS)

    MacKinnon, N. B.; Khojasteh, M.; Lane, P. M.; MacAulay, C. E.; Guillaud, M.; Stange, U.

    2007-02-01

    A series of hyperspectral transmission images of hematoxylin and eosin stained tissue sections from cervical biopsies were acquired at 10 nm intervals and assembled into a hyperspectral image cube. Custom software providing extraction of spectra at each pixel allows selection of images with maximum contrast for determination of selected features and differentiation of tissue features. Illumination profiles were created using a spectrally and temporally programmable light engine based on a spatial light modulator that can dynamically create any narrow or broadband spectral profile was used to select illumination wavelengths. Images were acquired with a monochrome CCD camera. Several methods of combining images from individual or composite spectral bands to recreate color images for pathologist review are shown. Unlike current "mechanical" illumination systems employing optical filters, filter wheels, motors, shutters and multiple control interfaces, the light engine integrates the lamp, wavelength control, intensity control and exposure control in a simple MEMS based system, where the only moving part is the lamp cooling fan. Illumination can now be programmed dynamically with digital control of all illumination parameters allowing wavelengths and intensities to be changed much faster than with filter wheels, and providing exposure control orders of magnitude more precise than mechanical shutters. This system can be integrated with digital imaging systems. Digitally controlled illumination is bit additive with image data providing high dynamic range imaging with monochrome or with color imaging devices. Performance of image analysis software for nuclear morphometric and tissue architecture analysis are compared for different wavelength regions.

  3. Instrumentation considerations in spectral imaging for tissue demarcation: comparing three methods of spectral resolution

    NASA Astrophysics Data System (ADS)

    Gebhart, Steven C.; Stokes, David L.; Vo-Dinh, Tuan; Mahadevan-Jansen, Anita

    2005-03-01

    Multiple methodologies exist to implement spectral imaging for tissue demarcation and disease diagnosis. In this paper, benchtop acousto-optic tunable filter (AOTF), liquid-crystal tunable filter (LCTF) and Fourier interferometric spectral imaging systems were quantitatively compared in terms of imaging speed of soft tissue autofluorescence. Optical throughput, image signal-to-noise ratio (SNR), and collagen autofluorescence imaging in chicken breast were assessed. Within this comparison, the Fourier system possessed the largest optical throughput (~50%) relative to the tunable-filter imaging systems; however, its throughput advantage failed to correlate to improved image SNR over the LCTF system. Further, while the autofluorescence imaging capability of the Fourier system exceeded that of the LCTF system for comparable total image integration times, the LCTF is capable of producing equivalent autofluorescence SNR with superior SNR when interrogations at only a few wavelengths are required and the random access filter tuning of the LCTF can be exploited. Therefore, the simple, rugged design and random-access filter-tuning capability of LCTF-based spectral imaging makes it best-suited for clinical development of soft tissue autofluorescence imaging.

  4. Learning Hierarchical Spectral-Spatial Features for Hyperspectral Image Classification.

    PubMed

    Zhou, Yicong; Wei, Yantao

    2016-07-01

    This paper proposes a spectral-spatial feature learning (SSFL) method to obtain robust features of hyperspectral images (HSIs). It combines the spectral feature learning and spatial feature learning in a hierarchical fashion. Stacking a set of SSFL units, a deep hierarchical model called the spectral-spatial networks (SSN) is further proposed for HSI classification. SSN can exploit both discriminative spectral and spatial information simultaneously. Specifically, SSN learns useful high-level features by alternating between spectral and spatial feature learning operations. Then, kernel-based extreme learning machine (KELM), a shallow neural network, is embedded in SSN to classify image pixels. Extensive experiments are performed on two benchmark HSI datasets to verify the effectiveness of SSN. Compared with state-of-the-art methods, SSN with a deep hierarchical architecture obtains higher classification accuracy in terms of the overall accuracy, average accuracy, and kappa ( κ ) coefficient of agreement, especially when the number of the training samples is small.

  5. Microscopic spectral imaging using mid-infrared semiconductor lasers

    NASA Astrophysics Data System (ADS)

    Guo, Bujin-.; Wang, Yi; Peng, Chuan; Luo, Guipeng; Le, Han Q.

    2003-07-01

    Infrared micro-spectroscopy is a useful tool for basic research and biomedical applications. Conventional microspectroscopic imaging apparatuses use thermal sources for sample illumination, which have low brightness, low optical spectral intensity, and high noise. This work evaluates the system engineering advantages of using mid-infrared semiconductor lasers that offer orders-of magnitude higher brightness, spectral intensity, and lower noise. A laser-based microscopic spectral imaging system with focal plane array detectors demonstrated a high signal-to-noise ratio (>20 dB) at video frame rate for a large illuminated area. Microscopic spectral imaging with fixed-wavelength and tunable lasers of 4.6, 6, and 9.3-μm wavelength was applied to a number of representative samples that consist of biological tissues (plant and animal) and solid material (a stack of laminated polymers). Transmission spectral images with ~30-dB dynamic range were obtained with clear evidence of spectral features for different samples. The potential of more advanced systems with a wide coverage of spectral bands is discussed.

  6. Spectral/Image Data Cubes of Europa, Ganymede, and Callisto

    NASA Astrophysics Data System (ADS)

    Granahan, J. C.; Polk, K. S.; Fanale, F. P.; McCord, T. B.

    1996-03-01

    A set of spectral image cubes of Europa, Ganymede, and Callisto have been constructed from Voyager mission images in an effort to prepare for the Jovian system portion of the Galileo spacecraft mission. Unfortunately, due to the fact that this spacecraft is presently restricted by an optimal 40 bits per second downlink and a finite onboard tape data storage many general observations of these moons will be restricted. Galileo imaging experiments like the Solid State Imager (SSI) and the Near Infrared Mapping Spectrometer (NIMS) will be restricted to making specific targeted observations due to their data intensive usage of spacecraft downlink and tape storage. These spectral image cubes are designed to provide complementary spectral and spatial coverage of Europa, Ganymede, and Callisto utilizing up to date control network measurements and photometric correction algorithms.

  7. Elucidation of the mechanisms of optical clearing in collagen tissue with multiphoton imaging.

    PubMed

    Hovhannisyan, Vladimir; Hu, Po-Sheng; Chen, Shean-Jen; Kim, Chang-Seok; Dong, Chen-Yuan

    2013-04-01

    Optical clearing (OC) is a promising method to overcome limitations in biomedical depth-resolved optical studies. Mechanisms of OC in purified bovine Achilles tendon, chicken skin, and chicken tendon were studied using time-lapsed, three-dimensional second harmonic generation (SHG) and two-photon fluorescence microscopic imaging. Quantified nonlinear optical measurements allowed temporal separation of two processes in collagen OC with glycerol. The first one is a fast process of tissue dehydration accompanied with collagen shrinkage and the second relatively slow process is glycerol penetration into the interfibrillar space of collagen alongside with CF swelling. The use of 50% glycerol induced less-expressed OC via partial substitution of water molecules with glycerol molecules. We also found that phosphate-buffered saline- and glycerol-treatments were reversible, and fiber morphology and SHG signal intensity were recovered after the removal of immersion agents. It was shown that tissue OC was a dynamic process and elucidation of its physical mechanisms may help choose optimal diagnostic, treatment, and modification regimes for collagen-based as well as other types of biomaterials. PMID:23552636

  8. Elucidation of the mechanisms of optical clearing in collagen tissue with multiphoton imaging.

    PubMed

    Hovhannisyan, Vladimir; Hu, Po-Sheng; Chen, Shean-Jen; Kim, Chang-Seok; Dong, Chen-Yuan

    2013-04-01

    Optical clearing (OC) is a promising method to overcome limitations in biomedical depth-resolved optical studies. Mechanisms of OC in purified bovine Achilles tendon, chicken skin, and chicken tendon were studied using time-lapsed, three-dimensional second harmonic generation (SHG) and two-photon fluorescence microscopic imaging. Quantified nonlinear optical measurements allowed temporal separation of two processes in collagen OC with glycerol. The first one is a fast process of tissue dehydration accompanied with collagen shrinkage and the second relatively slow process is glycerol penetration into the interfibrillar space of collagen alongside with CF swelling. The use of 50% glycerol induced less-expressed OC via partial substitution of water molecules with glycerol molecules. We also found that phosphate-buffered saline- and glycerol-treatments were reversible, and fiber morphology and SHG signal intensity were recovered after the removal of immersion agents. It was shown that tissue OC was a dynamic process and elucidation of its physical mechanisms may help choose optimal diagnostic, treatment, and modification regimes for collagen-based as well as other types of biomaterials.

  9. Elucidation of the mechanisms of optical clearing in collagen tissue with multiphoton imaging

    NASA Astrophysics Data System (ADS)

    Hovhannisyan, Vladimir; Hu, Po-Sheng; Chen, Shean-Jen; Kim, Chang-Seok; Dong, Chen-Yuan

    2013-04-01

    Optical clearing (OC) is a promising method to overcome limitations in biomedical depth-resolved optical studies. Mechanisms of OC in purified bovine Achilles tendon, chicken skin, and chicken tendon were studied using time-lapsed, three-dimensional second harmonic generation (SHG) and two-photon fluorescence microscopic imaging. Quantified nonlinear optical measurements allowed temporal separation of two processes in collagen OC with glycerol. The first one is a fast process of tissue dehydration accompanied with collagen shrinkage and the second relatively slow process is glycerol penetration into the interfibrillar space of collagen alongside with CF swelling. The use of 50% glycerol induced less-expressed OC via partial substitution of water molecules with glycerol molecules. We also found that phosphate-buffered saline- and glycerol-treatments were reversible, and fiber morphology and SHG signal intensity were recovered after the removal of immersion agents. It was shown that tissue OC was a dynamic process and elucidation of its physical mechanisms may help choose optimal diagnostic, treatment, and modification regimes for collagen-based as well as other types of biomaterials.

  10. Hyperspectral stimulated Raman scattering and multiphoton imaging for digital pathology of colonic disease

    NASA Astrophysics Data System (ADS)

    Wang, Zi; Zheng, Wei; Lin, Jian; Huang, Zhiwei

    2016-03-01

    Histopathology examinations of H&E stained biopsied tissues is the golden standard for colonic diseases (e.g., polyps, adenoma, and adenocarcinoma) diagnosis. However, staining effect of sample and doctor's expertise degree may greatly influence the diagnosis results. The information provided by the H&E stained sample is also limited to the morphological and PH information and no quantative information is available. In this paper, we report the development of a unique multimodal nonlinear optical microscopy (i.e., hyperspectral stimulated Raman scattering (hsSRS), second-harmonic generation (SHG), third-harmonic generation (THG), two-photon excitation fluorescence (TPEF)) platform for the diagnosis and characterization of colonic diseases. HsSRS in both fingerprint (800-1800 cm-1) and high-wavenumber (2800-3600 cm-1) regions allows us to discriminate different constituents with tiny difference in the Raman spectra. The increase of proteins and reduction of lipids could be observed with the progress of colonic cancer. SHG shows the distribution of collagen, which is found to aggregate for adenocarcinoma. TPEF provides the cell morphological and can reflect the damage inside glands caused by the diseases. THG shows the increase of optical heterogeneity related to cancer process. This work shows that the integrated hsSRS and TPEF/SHG/THG imaging technique can be an effective method for digital pathology of colonic diseases at the molecular and sub-cellular levels.

  11. Spectral image reconstruction by a tunable LED illumination

    NASA Astrophysics Data System (ADS)

    Lin, Meng-Chieh; Tsai, Chen-Wei; Tien, Chung-Hao

    2013-09-01

    Spectral reflectance estimation of an object via low-dimensional snapshot requires both image acquisition and a post numerical estimation analysis. In this study, we set up a system incorporating a homemade cluster of LEDs with spectral modulation for scene illumination, and a multi-channel CCD to acquire multichannel images by means of fully digital process. Principal component analysis (PCA) and pseudo inverse transformation were used to reconstruct the spectral reflectance in a constrained training set, such as Munsell and Macbeth Color Checker. The average reflectance spectral RMS error from 34 patches of a standard color checker were 0.234. The purpose is to investigate the use of system in conjunction with the imaging analysis for industry or medical inspection in a fast and acceptable accuracy, where the approach was preliminary validated.

  12. Methods for spectral image analysis by exploiting spatial simplicity

    DOEpatents

    Keenan, Michael R.

    2010-05-25

    Several full-spectrum imaging techniques have been introduced in recent years that promise to provide rapid and comprehensive chemical characterization of complex samples. One of the remaining obstacles to adopting these techniques for routine use is the difficulty of reducing the vast quantities of raw spectral data to meaningful chemical information. Multivariate factor analysis techniques, such as Principal Component Analysis and Alternating Least Squares-based Multivariate Curve Resolution, have proven effective for extracting the essential chemical information from high dimensional spectral image data sets into a limited number of components that describe the spectral characteristics and spatial distributions of the chemical species comprising the sample. There are many cases, however, in which those constraints are not effective and where alternative approaches may provide new analytical insights. For many cases of practical importance, imaged samples are "simple" in the sense that they consist of relatively discrete chemical phases. That is, at any given location, only one or a few of the chemical species comprising the entire sample have non-zero concentrations. The methods of spectral image analysis of the present invention exploit this simplicity in the spatial domain to make the resulting factor models more realistic. Therefore, more physically accurate and interpretable spectral and abundance components can be extracted from spectral images that have spatially simple structure.

  13. Methods for spectral image analysis by exploiting spatial simplicity

    DOEpatents

    Keenan, Michael R.

    2010-11-23

    Several full-spectrum imaging techniques have been introduced in recent years that promise to provide rapid and comprehensive chemical characterization of complex samples. One of the remaining obstacles to adopting these techniques for routine use is the difficulty of reducing the vast quantities of raw spectral data to meaningful chemical information. Multivariate factor analysis techniques, such as Principal Component Analysis and Alternating Least Squares-based Multivariate Curve Resolution, have proven effective for extracting the essential chemical information from high dimensional spectral image data sets into a limited number of components that describe the spectral characteristics and spatial distributions of the chemical species comprising the sample. There are many cases, however, in which those constraints are not effective and where alternative approaches may provide new analytical insights. For many cases of practical importance, imaged samples are "simple" in the sense that they consist of relatively discrete chemical phases. That is, at any given location, only one or a few of the chemical species comprising the entire sample have non-zero concentrations. The methods of spectral image analysis of the present invention exploit this simplicity in the spatial domain to make the resulting factor models more realistic. Therefore, more physically accurate and interpretable spectral and abundance components can be extracted from spectral images that have spatially simple structure.

  14. Spectral characteristics of two-photon autofluorescence and second harmonic generation from human skin in vivo

    NASA Astrophysics Data System (ADS)

    Breunig, Hans G.; König, Karsten

    2011-03-01

    We performed multiphoton imaging of human skin and recorded in combination the complete spectral content of the signals in vivo. The spectra represent the integration of multiphoton signals over the investigated regions of the epidermis and dermis. They are used to study depth-resolved in vivo emission characteristics of main endogenous skin fluorophores like keratin, NAD(P)H, collagen and elastin. The identification of the specific fluorophores is supported by analysis of additional in vivo fluorescence lifetime imaging. Furthermore, as a potential application of spectrally selective imaging the possibility to investigate the penetration of nanoparticles from sunscreen lotion into skin in vivo is discussed.

  15. Multiphoton microscopy of atheroslcerotic plaques

    NASA Astrophysics Data System (ADS)

    Lilledahl, Magnus B.; de Lange Davies, Catharina; Haugen, Olav A.; Svaasand, Lars O.

    2007-02-01

    Multiphoton microscopy is a techniques that fascilitates three dimensional imaging of intact, unstained tissue. Especially connective tissue has a relatively strong nonlinear optical response and can easily be imaged. Atherosclerosis is a disease where lipids accumulate in the vessel wall and there is a thickening of the intima by growth of a cap of connective tissue. The mechanical strength of this fibrous cap is of clinically importance. If the cap ruptures a thrombosis forms which can block a coronary vessel and therby causing myocardial infarction. Multiphoton microscopy can be used to image the fibrous cap and thereby determine the thickness of the cap and the structure of the connective fibres. This could possibly be developed into a diagnostic and clincal tool to monitor the vulnerability of a plaque and also to better understand the development of a plaque and effects of treatment. We have collected multiphoton microscopy images from atherosclerotic plaque in human aorta, both two photon excited fluorescens and second harmonic generated signal. The feasability of using this technique to determine the state of the plaque is explored.

  16. Hyperspectral image-based methods for spectral diversity

    NASA Astrophysics Data System (ADS)

    Sotomayor, Alejandro; Medina, Ollantay; Chinea, J. D.; Manian, Vidya

    2015-05-01

    Hyperspectral images are an important tool to assess ecosystem biodiversity. To obtain more precise analysis of biodiversity indicators that agree with indicators obtained using field data, analysis of spectral diversity calculated from images have to be validated with field based diversity estimates. The plant species richness is one of the most important indicators of biodiversity. This indicator can be measured in hyperspectral images considering the Spectral Variation Hypothesis (SVH) which states that the spectral heterogeneity is related to spatial heterogeneity and thus to species richness. The goal of this research is to capture spectral heterogeneity from hyperspectral images for a terrestrial neo tropical forest site using Vector Quantization (VQ) method and then use the result for prediction of plant species richness. The results are compared with that of Hierarchical Agglomerative Clustering (HAC). The validation of the process index is done calculating the Pearson correlation coefficient between the Shannon entropy from actual field data and the Shannon entropy computed in the images. One of the advantages of developing more accurate analysis tools would be the extension of the analysis to larger zones. Multispectral image with a lower spatial resolution has been evaluated as a prospective tool for spectral diversity.

  17. Advances in Spectral-Spatial Classification of Hyperspectral Images

    NASA Technical Reports Server (NTRS)

    Fauvel, Mathieu; Tarabalka, Yuliya; Benediktsson, Jon Atli; Chanussot, Jocelyn; Tilton, James C.

    2012-01-01

    Recent advances in spectral-spatial classification of hyperspectral images are presented in this paper. Several techniques are investigated for combining both spatial and spectral information. Spatial information is extracted at the object (set of pixels) level rather than at the conventional pixel level. Mathematical morphology is first used to derive the morphological profile of the image, which includes characteristics about the size, orientation and contrast of the spatial structures present in the image. Then the morphological neighborhood is defined and used to derive additional features for classification. Classification is performed with support vector machines using the available spectral information and the extracted spatial information. Spatial post-processing is next investigated to build more homogeneous and spatially consistent thematic maps. To that end, three presegmentation techniques are applied to define regions that are used to regularize the preliminary pixel-wise thematic map. Finally, a multiple classifier system is defined to produce relevant markers that are exploited to segment the hyperspectral image with the minimum spanning forest algorithm. Experimental results conducted on three real hyperspectral images with different spatial and spectral resolutions and corresponding to various contexts are presented. They highlight the importance of spectral-spatial strategies for the accurate classification of hyperspectral images and validate the proposed methods.

  18. Hyperspectral image segmentation using spatial-spectral graphs

    NASA Astrophysics Data System (ADS)

    Gillis, David B.; Bowles, Jeffrey H.

    2012-06-01

    Spectral graph theory has proven to be a useful tool in the analysis of high-dimensional data sets. Recall that, mathematically, a graph is a collection of objects (nodes) and connections between them (edges); a weighted graph additionally assigns numerical values (weights) to the edges. Graphs are represented by their adjacency whose elements are the weights between the nodes. Spectral graph theory uses the eigendecomposition of the adjacency matrix (or, more generally, the Laplacian of the graph) to derive information about the underlying graph. In this paper, we develop a spectral method based on the 'normalized cuts' algorithm to segment hyperspectral image data (HSI). In particular, we model an image as a weighted graph whose nodes are the image pixels, and edges defined as connecting spatial neighbors; the edge weights are given by a weighted combination of the spatial and spectral distances between nodes. We then use the Laplacian of the graph to recursively segment the image. The advantages of our approach are that, first, the graph structure naturally incorporates both the spatial and spectral information present in HSI; also, by using only spatial neighbors, the adjacency matrix is highly sparse; as a result, it is possible to apply our technique to much larger images than previous techniques. In the paper, we present the details of our algorithm, and include experimental results from a variety of hyperspectral images.

  19. Extraction of topographic and spectral albedo information from multispectral images.

    USGS Publications Warehouse

    Eliason, P.T.; Soderblom, L.A.; Chavez, P.A., Jr.

    1981-01-01

    A technique has been developed to separate and extract spectral-reflectivity variations and topographic informaiton from multispectral images. The process is a completely closed system employing only the image data and can be applied to any digital multispectral data set. -from Authors

  20. Multi-spectral confocal microendoscope for in-vivo imaging

    NASA Astrophysics Data System (ADS)

    Rouse, Andrew Robert

    The concept of in-vivo multi-spectral confocal microscopy is introduced. A slit-scanning multi-spectral confocal microendoscope (MCME) was built to demonstrate the technique. The MCME employs a flexible fiber-optic catheter coupled to a custom built slit-scan confocal microscope fitted with a custom built imaging spectrometer. The catheter consists of a fiber-optic imaging bundle linked to a miniature objective and focus assembly. The design and performance of the miniature objective and focus assembly are discussed. The 3mm diameter catheter may be used on its own or routed though the instrument channel of a commercial endoscope. The confocal nature of the system provides optical sectioning with 3mum lateral resolution and 30mum axial resolution. The prism based multi-spectral detection assembly is typically configured to collect 30 spectral samples over the visible chromatic range. The spectral sampling rate varies from 4nm/pixel at 490nm to 8nm/pixel at 660nm and the minimum resolvable wavelength difference varies from 7nm to 18nm over the same spectral range. Each of these characteristics are primarily dictated by the dispersive power of the prism. The MCME is designed to examine cellular structures during optical biopsy and to exploit the diagnostic information contained within the spectral domain. The primary applications for the system include diagnosis of disease in the gastro-intestinal tract and female reproductive system. Recent data from the grayscale imaging mode are presented. Preliminary multi-spectral results from phantoms, cell cultures, and excised human tissue are presented to demonstrate the potential of in-vivo multi-spectral imaging.

  1. Colorimetric-spectral clustering: a tool for multispectral image compression

    NASA Astrophysics Data System (ADS)

    Ciprian, R.; Carbucicchio, M.

    2011-11-01

    In this work a new compression method for multispectral images has been proposed: the 'colorimetric-spectral clustering'. The basic idea arises from the well-known cluster analysis, a multivariate analysis which finds the natural links between objects grouping them into clusters. In the colorimetric-spectral clustering compression method, the objects are the spectral reflectance factors of the multispectral images that are grouped into clusters on the basis of their colour difference. In particular two spectra can belong to the same cluster only if their colour difference is lower than a threshold fixed before starting the compression procedure. The performance of the colorimetric-spectral clustering has been compared to the k-means cluster analysis, in which the Euclidean distance between spectra is considered, to the principal component analysis and to the LabPQR method. The colorimetric-spectral clustering is able to preserve both the spectral and the colorimetric information of a multispectral image, allowing this information to be reproduced for all pixels of the image.

  2. Multi-spectral imaging with mid-infrared semiconductor lasers

    NASA Astrophysics Data System (ADS)

    Wang, Yi; Wang, Yang; Le, Han Q.

    2006-01-01

    Multi-spectral laser imaging can be a useful technology for target discrimination, classification, and identification based on object spectral signatures. The mid-IR region (~3-14 μm) is particularly rich of molecular spectroscopic fingerprints, but the technology has been under utilized. Compact, potentially inexpensive semiconductor lasers may allow more cost-effective applications. This paper describes a development of semiconductor-laser-based multi-spectral imaging for both near-IR and mid-IR, and demonstrates the potential of this technology. The near-IR study employed 7 wavelengths from 0.635-1.55 μm, and used for system engineering evaluation as well as for studying the fundamental aspects of multi-spectral laser imaging. These include issues of wavelength-dependence scattering as a function of incident and receiving angle and the polarization effects. Stokes vector imaging and degree-of-linear-polarization were shown to reveal significant information to characterize the targets. The mid-IR study employed 4 wavelengths from 3.3-9.6 μm, and was applied to diverse targets that consist of natural and man-made materials and household objects. It was shown capable to resolve and distinguish small spectral differences among various targets, thanks to the laser radiometric and spectral accuracy. Colorless objects in the visible were shown with "colorful" signatures in the mid-IR. An essential feature of the study is an advanced system architecture that employs wavelength-division-multiplexed laser beams for high spectral fidelity and resolution. In addition, unlike conventional one-transmitter and one receiver design, the system is based on a scalable CDMA network concept with multiple transmitters and receivers to allow efficient information acquisition. The results suggest that multi-spectral laser imaging in general can be a unique and powerful technology for wide ranging applications.

  3. Multiphoton processes: conference proceedings

    SciTech Connect

    Lambropoulos, P.; Smith, S.J.

    1984-01-01

    The chapters of this volume represent the invited papers delivered at the conference. They are arranged according to thermatic proximity beginning with atoms and continuing with molecules and surfaces. Section headings include multiphoton processes in atoms, field fluctuations and collisions in multiphoton process, and multiphoton processes in molecules and surfaces. Abstracts of individual items from the conference were prepared separately for the data base. (GHT)

  4. Tri-modal microscopy with multiphoton and optical coherence microscopy/tomography for multi-scale and multi-contrast imaging

    PubMed Central

    Chong, Shau Poh; Lai, Tom; Zhou, Yifeng; Tang, Shuo

    2013-01-01

    Multi-scale multimodal microscopy is a very useful technique by providing multiple imaging contrasts with adjustable field of views and spatial resolutions. Here, we present a tri-modal microscope combining multiphoton microscopy (MPM), optical coherence microscopy (OCM) and optical coherence tomography (OCT) for subsurface visualization of biological tissues. The advantages of the tri-modal system are demonstrated on various biological samples. It enables the visualization of multiple intrinsic contrasts including scattering, two-photon excitation fluorescence (TPEF), and second harmonic generation (SHG). It also enables a rapid scanning over a large tissue area and a high resolution zoom-in for cellular-level structures on regions of interest. The tri-modal microscope can be important for label-free imaging to obtain a sufficient set of parameters for reliable sample analysis. PMID:24049679

  5. New approach for the radiometric calibration of spectral imaging systems.

    PubMed

    Kohler, David; Bissett, W; Steward, Robert; Davis, Curtiss

    2004-05-31

    The calibration of multispectral and hyperspectral imaging systems is typically done in the laboratory using an integrating sphere, which usually produces a signal that is red rich. Using such a source to calibrate environmental monitoring systems presents some difficulties. Not only is much of the calibration data outside the range and spectral quality of data values that are expected to be captured in the field, using these measurements alone may exaggerate the optical flaws found within the system. Left unaccounted for, these flaws will become embedded in to the calibration, and thus, they will be passed on to the field data when the calibration is applied. To address these issues, we used a series of well-characterized spectral filters within our calibration. It provided us with a set us stable spectral standards to test and account for inadequacies in the spectral and radiometric integrity of the optical imager.

  6. Spatially resolved spectral-imaging device

    DOEpatents

    Bloom, Joshua Simon; Tyson, John Anthony

    2016-02-09

    A spatially resolved spectral device comprising a dispersive array to receive an incident light comprising a principal ray. The dispersive array comprising a plurality of dichroic layers, each of the plurality of dichroic layers disposed in a path of a direction of the principal ray. Each of the plurality of dichroic layers configured to at least one of reflect or transmit a different wavelength range of the incident light. The device further comprising a detection array operatively coupled with the dispersive array. The detection array comprising a photosensitive component including a plurality of detection pixels, each of the plurality of detection pixels having a light-receiving surface disposed parallel to the direction of the principal ray to detect a respective one of the different wavelength ranges of incident light reflected from a corresponding one of the plurality of dichroic layers.

  7. Multiphoton microscopy in defining liver function

    NASA Astrophysics Data System (ADS)

    Thorling, Camilla A.; Crawford, Darrell; Burczynski, Frank J.; Liu, Xin; Liau, Ian; Roberts, Michael S.

    2014-09-01

    Multiphoton microscopy is the preferred method when in vivo deep-tissue imaging is required. This review presents the application of multiphoton microscopy in defining liver function. In particular, multiphoton microscopy is useful in imaging intracellular events, such as mitochondrial depolarization and cellular metabolism in terms of NAD(P)H changes with fluorescence lifetime imaging microscopy. The morphology of hepatocytes can be visualized without exogenously administered fluorescent dyes by utilizing their autofluorescence and second harmonic generation signal of collagen, which is useful in diagnosing liver disease. More specific imaging, such as studying drug transport in normal and diseased livers are achievable, but require exogenously administered fluorescent dyes. If these techniques can be translated into clinical use to assess liver function, it would greatly improve early diagnosis of organ viability, fibrosis, and cancer.

  8. Identifying volcanic endmembers in hyperspectral images using spectral unmixing

    NASA Astrophysics Data System (ADS)

    Piscini, Alessandro; Carboni, Elisa; Del Frate, Fabio; Grainger, Roy Gordon

    2014-10-01

    Spectral unmixing technique is used in remote sensed data analysis for the determination of certain basis spectra called 'endmembers'. Once those spectra are found, the image cube can be 'unmixed' into fractional abundance of each material in each pixel. In the present work infrared spectra recorded by Infrared Atmospheric Sounding Interferometer (IASI) were used to characterize the emission from Grimsvotn volcanic eruption on 2011. In particular, a methodology based on spectral unmixing theory was used in order to extract the spectral signature of volcanic cloud constituents, such as ash and sulphur dioxide (SO2) and maps of their abundances in a IASI image were obtained. Taking the advantage of IASI broad spectral coverage the broadband signature in the Thermal Infrared (TIR) radiance spectra in the 1000-1410 cm-1 range associated with the presence of aerosols was obtained. Volcanic ash and SO2 spectral signatures were extracted, as well as those related to the simultaneous presence of ash, SO2 and cloud. The study proved that spectral unmixing, applied to Hyperspectral images, is able to identify volcanic aerosols and other species like SO2 despite a strong presence of meteorological clouds. Moreover, the analysis of hyperspectral datasets permitted to generate abundance maps for each endmember extracted. In particular, maps obtained for the test case of 2011 May, 23th put in evidence the separation between clouds of ejected SO2 and volcanic ash. The former dispersed at Northern latitudes, whilst the latter was situated at southern latitudes, South of Iceland.

  9. Time-resolved spectral imaging: better photon economy, higher accuracy

    NASA Astrophysics Data System (ADS)

    Fereidouni, Farzad; Reitsma, Keimpe; Blab, Gerhard A.; Gerritsen, Hans C.

    2015-03-01

    Lifetime and spectral imaging are complementary techniques that offer a non-invasive solution for monitoring metabolic processes, identifying biochemical compounds, and characterizing their interactions in biological tissues, among other tasks. Newly developed instruments that perform time-resolved spectral imaging can provide even more information and reach higher sensitivity than either modality alone. Here we report a multispectral lifetime imaging system based on a field-programmable gate array (FPGA), capable of operating at high photon count rates (12 MHz) per spectral detection channel, and with time resolution of 200 ps. We performed error analyses to investigate the effect of gate width and spectral-channel width on the accuracy of estimated lifetimes and spectral widths. Temporal and spectral phasors were used for analysis of recorded data, and we demonstrated blind un-mixing of the fluorescent components using information from both modalities. Fractional intensities, spectra, and decay curves of components were extracted without need for prior information. We further tested this approach with fluorescently doubly-labeled DNA, and demonstrated its suitability for accurately estimating FRET efficiency in the presence of either non-interacting or interacting donor molecules.

  10. Test and analysis of spectral response for UV image intensifier

    NASA Astrophysics Data System (ADS)

    Qian, Yunsheng; Liu, Jian; Feng, Cheng; Lv, Yang; Zhang, Yijun

    2015-10-01

    The UV image intensifier is one kind of electric vacuum imaging device based on principle of photoelectronic imaging. To achieve solar-blind detection, its spectral response characteristic is extremely desirable. A broad spectrum response measurement system is developed. This instrument uses EQ-99 laser-driven light source to get broad spectrum in the range of 200 nm to 1700 nm. A special preamplifier as well as a test software is work out. The spectral response of the image intensifier can be tested in the range of 200~1700 nm. Using this spectrum response measuring instrument, the UV image intensifiers are tested. The spectral response at the spectral range of 200 nm to 600 nm are obtained. Because of the quantum efficiency of Te-Cs photocathode used in image intens ifier above 280nm wavelength still exists, especially at 280 nm to 320nm.Therefore, high-performance UV filters is required for solar blind UV detection. Based on two sets of UV filters, the influence of solar radiation on solar blind detection is calculated and analyzed.

  11. Multi-spectral image dissector camera system

    NASA Technical Reports Server (NTRS)

    1972-01-01

    The image dissector sensor for the Earth Resources Program is evaluated using contrast and reflectance data. The ground resolution obtainable for low contrast at the targeted signal to noise ratio of 1.8 was defined. It is concluded that the system is capable of achieving the detection of small, low contrast ground targets from satellites.

  12. Spectral characterization of storage phosphor imaging plates

    SciTech Connect

    Deresch, A.; Jechow, M.; Gollwitzer, C.

    2014-02-18

    This work presents attenuation and sensitivity measurements of radiographic imaging plates (IPs) with quasimonoenergetic X-rays in the 8–60 kiloelectronvolt range. The measurements are used to validate theoretical predictions. A short overview of the theoretical model is given. The model can be used to describe the sensitivity of different detector types to a wide range of X-ray energies.

  13. Spectral image destriping using a low-dimensional model

    NASA Astrophysics Data System (ADS)

    Adler-Golden, S.; Richtsmeier, S.; Conforti, P.; Bernstein, L.

    2013-05-01

    Striping effects, i.e., artifacts that vary systematically with the image column or row, may arise in hyperspectral or multispectral imagery from a variety of sources. One potential source of striping is a physical effect inherent in the measurement, such as a variation in viewing geometry or illumination across the image. More common sources are instrumental artifacts, such as a variation in spectral resolution, wavelength calibration or radiometric calibration, which can result from imperfect corrections for spectral "smile" or detector array nonuniformity. This paper describes a general method of suppressing striping effects in spectral imagery by referencing the image to a spectrally lowdimensional model. The destriping transform for a given column or row is taken to be affine, i.e., specified by a gain and offset. The image cube model is derived from a subset of spectral bands or principal components thereof. The general approach is effective for all types of striping, including broad or narrow, sharp or graduated, and is applicable to radiance data at all optical wavelengths and to reflectance data in the solar (visible through short-wave infrared) wavelength region. Some specific implementations are described, including a method for suppressing effects of viewing angle variation in VNIR-SWIR imagery.

  14. Spectral imaging detects breast cancer in fresh unstained specimens

    NASA Astrophysics Data System (ADS)

    Chung, Alice; Gaon, Mark; Jeong, Jihoon; Karlan, Scott; Lindsley, Erik; Wachsmann-Hogiu, Sebastian; Xiong, Yizhi; Zhao, Tong; Farkas, Daniel L.

    2006-02-01

    Spectral imaging has recently been introduced in the biomedical field as a noninvasive, quantitative means of studying biological tissues. Many of its potential applications have been demonstrated (in vitro and, to a lesser degree, in vivo) with the use of stains or dyes. Successful translation to the clinical environment has been largely lagging, due to safety considerations and regulatory limitations preventing use of contrast agents in humans. We report experiments showing the feasibility of high-resolution spectral imaging of breast cancer without the use of contrast agents, thus completing the continuum of translational research, to in vivo imaging that will be directly applicable in the clinical environment. Our initial work focused on image acquisition using Fourier transform microinterferometry and subsequent segmentation of both stained and unstained breast cancer slides-derived image sets. We then applied our techniques to imaging fresh unstained ex vivo specimens of rat breast cancer and sentinel lymph nodes. We also investigated multiple methods of classification to optimize our image analyses, and preliminary results for the best algorithm tested yielded an overall sensitivity of 96%, and a specificity of 92% for cancer detection. Using spectral imaging and classification techniques, we were able to demonstrate that reliable detection of breast cancer in fixed and fresh unstained specimens of breast tissue is possible.

  15. Spectral Imaging for Intracranial Stents and Stent Lumen

    PubMed Central

    Chen, David Yen-Ting; Chen, Chi-Jen; Hsu, Hui-Ling

    2016-01-01

    Introduction Application of computed tomography for monitoring intracranial stents is limited because of stent-related artifacts. Our purpose was to evaluate the effect of gemstone spectral imaging on the intracranial stent and stent lumen. Materials and Methods In vitro, we scanned Enterprise stent phantom and a stent–cheese complex using the gemstone spectral imaging protocol. Follow-up gemstone spectral images of 15 consecutive patients with placement of Enterprise from January 2013 to September 2014 were also retrospectively reviewed. We used 70-keV, 140-keV, iodine (water), iodine (calcium), and iodine (hydroxyapatite) images to evaluate their effect on the intracranial stent and stent lumen. Two regions of interest were individually placed in stent lumen and adjacent brain tissue. Contrast-to-noise ratio was measured to determine image quality. The maximal diameter of stent markers was also measured to evaluate stent-related artifact. Two radiologists independently graded the visibility of the lumen at the maker location by using a 4-point scale. The mean of grading score, contrast/noise ratio and maximal diameter of stent markers were compared among all modes. All results were analyzed by SPSS version 20. Results In vitro, iodine (water) images decreased metallic artifact of stent makers to the greatest degree. The most areas of cheese were observed on iodine (water) images. In vivo, iodine (water) images had the smallest average diameter of stent markers (0.33 ± 0.17mm; P < .05) and showed the highest mean grading score (2.94 ± 0.94; P < .05) and contrast/noise ratio of in-stent lumen (160.03 ±37.79; P < .05) among all the modes. Conclusion Iodine (water) images can help reduce stent-related artifacts of Enterprise and enhance contrast of in-stent lumen. Spectral imaging may be considered a noninvasive modality for following-up patients with in-stent stenosis. PMID:26731534

  16. Automated spectral imaging for clinical diagnostics

    NASA Astrophysics Data System (ADS)

    Breneman, John; Heffelfinger, David M.; Pettipiece, Ken; Tsai, Chris; Eden, Peter; Greene, Richard A.; Sorensen, Karen J.; Stubblebine, Will; Witney, Frank

    1998-04-01

    Bio-Rad Laboratories supplies imaging equipment for many applications in the life sciences. As part of our effort to offer more flexibility to the investigator, we are developing a microscope-based imaging spectrometer for the automated detection and analysis of either conventionally or fluorescently labeled samples. Immediate applications will include the use of fluorescence in situ hybridization (FISH) technology. The field of cytogenetics has benefited greatly from the increased sensitivity of FISH producing simplified analysis of complex chromosomal rearrangements. FISH methods for identification lends itself to automation more easily than the current cytogenetics industry standard of G- banding, however, the methods are complementary. Several technologies have been demonstrated successfully for analyzing the signals from labeled samples, including filter exchanging and interferometry. The detection system lends itself to other fluorescent applications including the display of labeled tissue sections, DNA chips, capillary electrophoresis or any other system using color as an event marker. Enhanced displays of conventionally stained specimens will also be possible.

  17. High spectral resolution image of 'Stripe'

    NASA Technical Reports Server (NTRS)

    1997-01-01

    Another early target for the full-color capability of the Imager for Mars Pathfinder (IMP) was the rock Stripe, named for the vertical, red band on its face. Regions with different color properties include the rock face (blue), bright soil in the background (green), darker intermediate colored soil in front of the rock (red), and the vertical stripe.

    Mars Pathfinder is the second in NASA's Discovery program of low-cost spacecraft with highly focused science goals. The Jet Propulsion Laboratory, Pasadena, CA, developed and manages the Mars Pathfinder mission for NASA's Office of Space Science, Washington, D.C. The Imager for Mars Pathfinder (IMP) was developed by the University of Arizona Lunar and Planetary Laboratory under contract to JPL. Peter Smith is the Principal Investigator. JPL is an operating division of the California Institute of Technology (Caltech).

  18. Spectral imaging of mouse calvaria undergoing craniosynstosis

    NASA Astrophysics Data System (ADS)

    Crane, Nicole J.; Wang, Wei; Ignelzi, Michael A., Jr.; Morris, Michael D.

    2003-07-01

    Craniosynostosis, the premature fusion of the skull bones at the sutures, is the second most common human birth defect that affects the face and skull. The top most flat bones that comprise the skull, or calvaria, are most often affected. We previously showed that treatment of mouse calvaria with FGF2-soaked beads leads to craniosynostosis. In this study we treated mouse calvaria with FGF2-soaked beads and then used Raman imaging to demonstrate the spatial distribution of apatitic mineral and matrix in the sutures. There was no difference between FGF2 treated and control calvaria in the type of mineral produced (a lightly carbonated apatite), however we did observe increased mineral deposition in FGF2 treated calvaria. Raman imaging has great promise to detect the earliest mineral and matrix changes that occur in craniosynostosis.

  19. Some simple mechanisms of multiphoton excitation in many - level systems

    NASA Astrophysics Data System (ADS)

    Donley, E. A.; Marquardt, R.; Quack, M.; Stohner, J.; Thanopulos, I.; Wallenborn, E.-U.

    Results are reported on coherent monochromatic multiphoton excitation in many-level systems, which are representative for some of the basic mechanisms for atomic and molecular multiphoton processes. Numerical solutions are discussed that use the Floquet and quasiresonant approximations in the framework of the URIMIR program package. The excitation schemes include direct three-photon excitation, two-photon excitation with diagonal coupling, Göppert-Mayer-type two-photon processes, multiphoton excitation with off-resonant intermediates, and practically irreversible coherent excitation into dense spectral structures. Several interesting phenomena are observed, such as nonlinear line shifts and broadenings of multiphoton resonances of relevance for multiphoton spectroscopy and almost constant intermediate population inversions, potentially useful for laser design. The accurate numerical results are compared with approximate solutions from perturbation theory, and with simple analytical solutions from Rabi-type formulae.

  20. Molecular application of spectral photoacoustic imaging in pancreatic cancer pathology

    NASA Astrophysics Data System (ADS)

    Lakshman, Minalini; Hupple, Clinton; Lohse, Ines; Hedley, David; Needles, Andrew; Theodoropoulos, Catherine

    2012-12-01

    Spectral imaging is an advanced photo-acoustic (PA) mode that can discern optical absorption of contrast agent(s) in the tissue micro-environment. This advancement is made possible by precise control of optical wavelength using a tunable pulsed laser, ranging from 680-970 nm. Differential optical absorption of blood oxygenation states makes spectral imaging of hemoglobin ideal to investigate remodeling of the tumor microenvironment- a molecular change that renders resistance to standard cancer treatment. Approach: Photo-acoustic imaging was performed on the Vevo® LAZR system (VisualSonics) at 5-20 Hz. Deep abdominal imaging was accomplished with a LZ250D probe at a center frequency of 21MHz and an axial resolution of 75 μm. The tumor model was generated in an immune compromised mouse by surgical implantation of primary patient derived tumors, in the pancreas. Results: Spectral imaging for oxygen saturation at 750 nm and 850 nm characterized this tumor with a poorly oxygenated core surrounded by a well oxygenated periphery. Multispectral imaging identified a sub region in the core with a four-fold signal exclusively at 750 and 800 nm. A co-registered 2D image of this region was shown to be echogenic and calcification was suspected. Perfusion imaging with contrast enhanced ultrasound using microbubbles (Vevo MicroMarker® contrast agents, VisualSonics) identified functional vessels towards this sub region. Histology confirmed calcification and vascularization in the tumor core. Taken together, non-invasive characterization of the tumor microenvironment using photo-acoustics rendered spectral imaging a sensitive tool to monitor molecular changes representative of progression of pancreatic cancer that kills within 6 months of diagnosis.

  1. Surface science capabilities from IMP spectral imaging

    NASA Technical Reports Server (NTRS)

    Singer, Robert B.

    1994-01-01

    The Imager for Mars Pathfinder (IMP) had a single 12-position filter wheel for one of its two 'eyes'. Originally eight, and then nine, of these filters were optimized for surface science, and three narrow-band filters for atmospheric science. Because of some design revisions we will now have filter wheels on both sides. The wheels for right and left eyes are identical, 12 filter positions each, and rigidly linked to the same rotation shaft. There are now 13 surface filters, in addition to 5 for atmospheric observations. Details of all the filter positions are tabulated and approximate gaussian bandpasses for the 13 surface filters are shown.

  2. A Parametric Approach to Spectral-Spatial EPR Imaging

    PubMed Central

    Som, Subhojit; Potter, Lee C.; Ahmad, Rizwan; Kuppusamy, Periannan

    2007-01-01

    Continuous wave electron paramagnetic resonance imaging for in vivo mapping of spin distribution and spectral shape requires rapid data acquisition. A spectral-spatial imaging technique is presented that provides an order of magnitude reduction in acquisition time, compared to iterative tomographic reprojection. The proposed approach assumes that spectral shapes in the sample are well-approximated by members from a parametric family of functions. A model is developed for the spectra measured with magnetic field modulation. Parameters defining the spin distribution and spectral shapes are then determined directly from the measurements using maximum a posteriori probability estimation. The approach does not suffer approximation error from limited sweep width of the main magnetic field and explicitly incorporates the variability in signal-to-noise ratio versus strength of magnetic field gradient. The processing technique is experimentally demonstrated on a one- dimensional phantom containing a nitroxide spin label with constant g-factor. Using an L-band EPR spectrometer, spectral shapes and spin distribution are accurately recovered from two projections and a spectral window which is comparable to the maximum linewidth of the sample. PMID:17276111

  3. Spatial and spectral performance of a chromotomosynthetic hyperspectral imaging system.

    PubMed

    Bostick, Randall L; Perram, Glen P

    2012-03-01

    The spatial and spectral resolutions achievable by a prototype rotating prism chromotomosynthetic imaging (CTI) system operating in the visible spectrum are described. The instrument creates hyperspectral imagery by collecting a set of 2D images with each spectrally projected at a different rotation angle of the prism. Mathematical reconstruction techniques that have been well tested in the field of medical physics are used to reconstruct the data to produce the 3D hyperspectral image. The instrument operates with a 100 mm focusing lens in the spectral range of 400-900 nm with a field of view of 71.6 mrad and angular resolution of 0.8-1.6 μrad. The spectral resolution is 0.6 nm at the shortest wavelengths, degrading to over 10 nm at the longest wavelengths. Measurements using a point-like target show that performance is limited by chromatic aberration. The system model is slightly inaccurate due to poor estimation of detector spatial resolution, this is corrected based on results improving model performance. As with traditional dispersion technology, calibration of the transformed wavelength axis is required, though with this technology calibration improves both spectral and spatial resolution. While this prototype does not operate at high speeds, components exist which will allow for CTI systems to generate hyperspectral video imagery at rates greater than 100 Hz. PMID:22462909

  4. Spatial and spectral performance of a chromotomosynthetic hyperspectral imaging system

    NASA Astrophysics Data System (ADS)

    Bostick, Randall L.; Perram, Glen P.

    2012-03-01

    The spatial and spectral resolutions achievable by a prototype rotating prism chromotomosynthetic imaging (CTI) system operating in the visible spectrum are described. The instrument creates hyperspectral imagery by collecting a set of 2D images with each spectrally projected at a different rotation angle of the prism. Mathematical reconstruction techniques that have been well tested in the field of medical physics are used to reconstruct the data to produce the 3D hyperspectral image. The instrument operates with a 100 mm focusing lens in the spectral range of 400-900 nm with a field of view of 71.6 mrad and angular resolution of 0.8-1.6 μrad. The spectral resolution is 0.6 nm at the shortest wavelengths, degrading to over 10 nm at the longest wavelengths. Measurements using a point-like target show that performance is limited by chromatic aberration. The system model is slightly inaccurate due to poor estimation of detector spatial resolution, this is corrected based on results improving model performance. As with traditional dispersion technology, calibration of the transformed wavelength axis is required, though with this technology calibration improves both spectral and spatial resolution. While this prototype does not operate at high speeds, components exist which will allow for CTI systems to generate hyperspectral video imagery at rates greater than 100 Hz.

  5. Spectral prior image constrained compressed sensing (spectral PICCS) for photon-counting computed tomography

    NASA Astrophysics Data System (ADS)

    Yu, Zhicong; Leng, Shuai; Li, Zhoubo; McCollough, Cynthia H.

    2016-09-01

    Photon-counting computed tomography (PCCT) is an emerging imaging technique that enables multi-energy imaging with only a single scan acquisition. To enable multi-energy imaging, the detected photons corresponding to the full x-ray spectrum are divided into several subgroups of bin data that correspond to narrower energy windows. Consequently, noise in each energy bin increases compared to the full-spectrum data. This work proposes an iterative reconstruction algorithm for noise suppression in the narrower energy bins used in PCCT imaging. The algorithm is based on the framework of prior image constrained compressed sensing (PICCS) and is called spectral PICCS; it uses the full-spectrum image reconstructed using conventional filtered back-projection as the prior image. The spectral PICCS algorithm is implemented using a constrained optimization scheme with adaptive iterative step sizes such that only two tuning parameters are required in most cases. The algorithm was first evaluated using computer simulations, and then validated by both physical phantoms and in vivo swine studies using a research PCCT system. Results from both computer-simulation and experimental studies showed substantial image noise reduction in narrow energy bins (43-73%) without sacrificing CT number accuracy or spatial resolution.

  6. Spectral prior image constrained compressed sensing (spectral PICCS) for photon-counting computed tomography

    NASA Astrophysics Data System (ADS)

    Yu, Zhicong; Leng, Shuai; Li, Zhoubo; McCollough, Cynthia H.

    2016-09-01

    Photon-counting computed tomography (PCCT) is an emerging imaging technique that enables multi-energy imaging with only a single scan acquisition. To enable multi-energy imaging, the detected photons corresponding to the full x-ray spectrum are divided into several subgroups of bin data that correspond to narrower energy windows. Consequently, noise in each energy bin increases compared to the full-spectrum data. This work proposes an iterative reconstruction algorithm for noise suppression in the narrower energy bins used in PCCT imaging. The algorithm is based on the framework of prior image constrained compressed sensing (PICCS) and is called spectral PICCS; it uses the full-spectrum image reconstructed using conventional filtered back-projection as the prior image. The spectral PICCS algorithm is implemented using a constrained optimization scheme with adaptive iterative step sizes such that only two tuning parameters are required in most cases. The algorithm was first evaluated using computer simulations, and then validated by both physical phantoms and in vivo swine studies using a research PCCT system. Results from both computer-simulation and experimental studies showed substantial image noise reduction in narrow energy bins (43–73%) without sacrificing CT number accuracy or spatial resolution.

  7. Comparison of in vivo and ex vivo laser scanning microscopy and multiphoton tomography application for human and porcine skin imaging

    SciTech Connect

    Darvin, M E; Richter, H; Zhu, Y J; Meinke, M C; Knorr, F; Lademann, J; Gonchukov, S A; Koenig, K

    2014-07-31

    Two state-of-the-art microscopic optical methods, namely, confocal laser scanning microscopy in the fluorescence and reflectance regimes and multiphoton tomography in the autofluorescence and second harmonic generation regimes, are compared for porcine skin ex vivo and healthy human skin in vivo. All skin layers such as stratum corneum (SC), stratum spinosum (SS), stratum basale (SB), papillary dermis (PD) and reticular dermis (RD) as well as transition zones between these skin layers are measured noninvasively at a high resolution, using the above mentioned microscopic methods. In the case of confocal laser scanning microscopy (CLSM), measurements in the fluorescence regime were performed by using a fluorescent dye whose topical application on the surface is well suited for the investigation of superficial SC and characterisation of the skin barrier function. For investigations of deeply located skin layers, such as SS, SB and PD, the fluorescent dye must be injected into the skin, which markedly limits fluorescence measurements using CLSM. In the case of reflection CLSM measurements, the obtained results can be compared to the results of multiphoton tomography (MPT) for all skin layers excluding RD. CLSM cannot distinguish between dermal collagen and elastin measuring their superposition in the RD. By using MPT, it is possible to analyse the collagen and elastin structures separately, which is important for the investigation of anti-aging processes. The resolution of MPT is superior to CLSM. The advantages and limitations of both methods are discussed and the differences and similarities between human and porcine skin are highlighted. (laser biophotonics)

  8. Comparison of in vivo and ex vivo laser scanning microscopy and multiphoton tomography application for human and porcine skin imaging

    NASA Astrophysics Data System (ADS)

    Darvin, M. E.; Richter, H.; Zhu, Y. J.; Meinke, M. C.; Knorr, F.; Gonchukov, S. A.; Koenig, K.; Lademann, J.

    2014-07-01

    Two state-of-the-art microscopic optical methods, namely, confocal laser scanning microscopy in the fluorescence and reflectance regimes and multiphoton tomography in the autofluorescence and second harmonic generation regimes, are compared for porcine skin ex vivo and healthy human skin in vivo. All skin layers such as stratum corneum (SC), stratum spinosum (SS), stratum basale (SB), papillary dermis (PD) and reticular dermis (RD) as well as transition zones between these skin layers are measured noninvasively at a high resolution, using the above mentioned microscopic methods. In the case of confocal laser scanning microscopy (CLSM), measurements in the fluorescence regime were performed by using a fluorescent dye whose topical application on the surface is well suited for the investigation of superficial SC and characterisation of the skin barrier function. For investigations of deeply located skin layers, such as SS, SB and PD, the fluorescent dye must be injected into the skin, which markedly limits fluorescence measurements using CLSM. In the case of reflection CLSM measurements, the obtained results can be compared to the results of multiphoton tomography (MPT) for all skin layers excluding RD. CLSM cannot distinguish between dermal collagen and elastin measuring their superposition in the RD. By using MPT, it is possible to analyse the collagen and elastin structures separately, which is important for the investigation of anti-aging processes. The resolution of MPT is superior to CLSM. The advantages and limitations of both methods are discussed and the differences and similarities between human and porcine skin are highlighted.

  9. Information-Efficient Spectral Imaging Sensor With Tdi

    DOEpatents

    Rienstra, Jeffrey L.; Gentry, Stephen M.; Sweatt, William C.

    2004-01-13

    A programmable optical filter for use in multispectral and hyperspectral imaging employing variable gain time delay and integrate arrays. A telescope focuses an image of a scene onto at least one TDI array that is covered by a multispectral filter that passes separate bandwidths of light onto the rows in the TDI array. The variable gain feature of the TDI array allows individual rows of pixels to be attenuated individually. The attenuations are functions of the magnitudes of the positive and negative components of a spectral basis vector. The spectral basis vector is constructed so that its positive elements emphasize the presence of a target and its negative elements emphasize the presence of the constituents of the background of the imaged scene. This system provides for a very efficient determination of the presence of the target, as opposed to the very data intensive data manipulations that are required in conventional hyperspectral imaging systems.

  10. Content-based hyperspectral image retrieval using spectral unmixing

    NASA Astrophysics Data System (ADS)

    Plaza, Antonio J.

    2011-11-01

    The purpose of content-based image retrieval (CBIR) is to retrieve, from real data stored in a database, information that is relevant to a query. A major challenge for the development of efficient CBIR systems in the context of hyperspectral remote sensing applications is how to deal with the extremely large volumes of data produced by current Earth-observing (EO) imaging spectrometers. The data resulting from EO campaigns often comprises many Gigabytes per flight. When multiple instruments or timelines are combined, this leads to the collection of massive amounts of data coming from heterogeneous sources, and these data sets need to be effectively stored, managed, shared and retrieved. Furthermore, the growth in size and number of hyperspectral data archives demands more sophisticated search capabilities to allow users to locate and reuse data acquired in the past. In this paper we develop a new strategy to effectively retrieve hyperspectral image data sets using spectral unmixing concepts. Spectral unmixing is a very important task for hyperspectral data exploitation since the spectral signatures collected in natural environments are invariably a mixture of the pure signatures of the various materials found within the spatial extent of the ground instantaneous field view of the imaging instrument. In this work, we use the information provided by spectral unmixing (i.e. the spectral endmembers and their corresponding abundances in the scene) as effective meta-data to develop a new CBIR system that can assist users in the task of efficiently searching hyperspectral image instances in large data repositories. The proposed approach is validated using a collection of 154 hyperspectral data sets (comprising seven full flightlines) gathered by NASA using the Airborne Visible Infra-Red Imaging Spectrometer (AVIRIS) over the World Trade Center (WTC) area in New York City during the last two weeks of September, 2001, only a few days after the terrorist attacks that

  11. Spatial, Temporal and Spectral Satellite Image Fusion via Sparse Representation

    NASA Astrophysics Data System (ADS)

    Song, Huihui

    Remote sensing provides good measurements for monitoring and further analyzing the climate change, dynamics of ecosystem, and human activities in global or regional scales. Over the past two decades, the number of launched satellite sensors has been increasing with the development of aerospace technologies and the growing requirements on remote sensing data in a vast amount of application fields. However, a key technological challenge confronting these sensors is that they tradeoff between spatial resolution and other properties, including temporal resolution, spectral resolution, swath width, etc., due to the limitations of hardware technology and budget constraints. To increase the spatial resolution of data with other good properties, one possible cost-effective solution is to explore data integration methods that can fuse multi-resolution data from multiple sensors, thereby enhancing the application capabilities of available remote sensing data. In this thesis, we propose to fuse the spatial resolution with temporal resolution and spectral resolution, respectively, based on sparse representation theory. Taking the study case of Landsat ETM+ (with spatial resolution of 30m and temporal resolution of 16 days) and MODIS (with spatial resolution of 250m ~ 1km and daily temporal resolution) reflectance, we propose two spatial-temporal fusion methods to combine the fine spatial information of Landsat image and the daily temporal resolution of MODIS image. Motivated by that the images from these two sensors are comparable on corresponding bands, we propose to link their spatial information on available Landsat- MODIS image pair (captured on prior date) and then predict the Landsat image from the MODIS counterpart on prediction date. To well-learn the spatial details from the prior images, we use a redundant dictionary to extract the basic representation atoms for both Landsat and MODIS images based on sparse representation. Under the scenario of two prior Landsat

  12. SAR imaging via modern 2-D spectral estimation methods.

    PubMed

    DeGraaf, S R

    1998-01-01

    This paper discusses the use of modern 2D spectral estimation algorithms for synthetic aperture radar (SAR) imaging. The motivation for applying power spectrum estimation methods to SAR imaging is to improve resolution, remove sidelobe artifacts, and reduce speckle compared to what is possible with conventional Fourier transform SAR imaging techniques. This paper makes two principal contributions to the field of adaptive SAR imaging. First, it is a comprehensive comparison of 2D spectral estimation methods for SAR imaging. It provides a synopsis of the algorithms available, discusses their relative merits for SAR imaging, and illustrates their performance on simulated and collected SAR imagery. Some of the algorithms presented or their derivations are new, as are some of the insights into or analyses of the algorithms. Second, this work develops multichannel variants of four related algorithms, minimum variance method (MVM), reduced-rank MVM (RRMVM), adaptive sidelobe reduction (ASR) and space variant apodization (SVA) to estimate both reflectivity intensity and interferometric height from polarimetric displaced-aperture interferometric data. All of these interferometric variants are new. In the interferometric contest, adaptive spectral estimation can improve the height estimates through a combination of adaptive nulling and averaging. Examples illustrate that MVM, ASR, and SVA offer significant advantages over Fourier methods for estimating both scattering intensity and interferometric height, and allow empirical comparison of the accuracies of Fourier, MVM, ASR, and SVA interferometric height estimates.

  13. Advances in Hyperspectral and Multispectral Image Fusion and Spectral Unmixing

    NASA Astrophysics Data System (ADS)

    Lanaras, C.; Baltsavias, E.; Schindler, K.

    2015-08-01

    In this work, we jointly process high spectral and high geometric resolution images and exploit their synergies to (a) generate a fused image of high spectral and geometric resolution; and (b) improve (linear) spectral unmixing of hyperspectral endmembers at subpixel level w.r.t. the pixel size of the hyperspectral image. We assume that the two images are radiometrically corrected and geometrically co-registered. The scientific contributions of this work are (a) a simultaneous approach to image fusion and hyperspectral unmixing, (b) enforcing several physically plausible constraints during unmixing that are all well-known, but typically not used in combination, and (c) the use of efficient, state-of-the-art mathematical optimization tools to implement the processing. The results of our joint fusion and unmixing has the potential to enable more accurate and detailed semantic interpretation of objects and their properties in hyperspectral and multispectral images, with applications in environmental mapping, monitoring and change detection. In our experiments, the proposed method always improves the fusion compared to competing methods, reducing RMSE between 4% and 53%.

  14. Hyperspectral imaging of bruises in the SWIR spectral region

    NASA Astrophysics Data System (ADS)

    Randeberg, Lise L.; Hernandez-Palacios, Julio

    2012-02-01

    Optical diagnostics of bruised skin might provide important information for characterization and age determination of such injuries. Hyperspectral imaging is one of the optical techniques that have been employed for bruise characterization. This technique combines high spatial and spectral resolution and makes it possible to study both chromophore signatures and -distributions in an injury. Imaging and spectroscopy in the visible spectral range have resulted in increased knowledge about skin bruises. So far the SWIR region has not been explored for this application. The main objective of the current study was to characterize bruises in the SWIR wavelength range. Hyperspectral images in the SWIR (950-2500nm ) and VNIR (400-850nm) spectral range were collected from 3 adult volunteers with bruises of known age. Data were collected over a period of 8 days. The data were analyzed using spectroscopic techniques and statistical image analysis. Preliminary results from the pilot study indicate that SWIR hyperspectral imaging might be an important supplement to imaging in the visible part of the spectrum. The technique emphasizes local edema and gives a possibility to visualize features that cannot easily be seen in the visible part of the spectrum.

  15. Digital spectral imaging for histopathology and cytopathology

    NASA Astrophysics Data System (ADS)

    Levenson, Richard M.; Farkas, Daniel L.

    1997-05-01

    The process by which diseases, particularly neoplastic diseases, are diagnosed by pathologists using microscopic evaluation of tissue has changed little over the last several decades despite the advent of molecular medicine. Cells or tissues, stained with complex and sometimes poorly characterized organic dyes, are examined using a subjective, pattern-matching approach whose accuracy and reproducibility has increasingly been challenged. Furthermore, the ability of pathologists to deliver accurate prognoses using histological and clinical parameters remains limited, leading to both over- and under-treatment. While molecular techniques hold out great promise as diagnostic and prognostic tools, they are currently still largely investigational, and costly. A complementary approach is proposed, whereby more information is obtained by improved analysis of conventionally prepared histological and cytological samples. Using inverse Fourier transform multi- pixel spectroscopy, a new instrument has been developed which can display a complete transmittance or emission spectrum at every pixel of an image, providing much more color information than can be appreciated by eye or by conventional red-green-blue color cameras. Since spectra variations in staining behavior correlate with alterations in subcellular macromolecular composition it seems likely that they may also correlate with diagnosis and clinical behavior for a number of disease states. Examples of how this approach may prove useful in clinical practice are provided.

  16. Lossless compression of multispectral images using spectral information

    NASA Astrophysics Data System (ADS)

    Ma, Long; Shi, Zelin; Tang, Xusheng

    2009-10-01

    Multispectral images are available for different purposes due to developments in spectral imaging systems. The sizes of multispectral images are enormous. Thus transmission and storage of these volumes of data require huge time and memory resources. That is why compression algorithms must be developed. A salient property of multispectral images is that strong spectral correlation exists throughout almost all bands. This fact is successfully used to predict each band based on the previous bands. We propose to use spectral linear prediction and entropy coding with context modeling for encoding multispectral images. Linear prediction predicts the value for the next sample and computes the difference between predicted value and the original value. This difference is usually small, so it can be encoded with less its than the original value. The technique implies prediction of each image band by involving number of bands along the image spectra. Each pixel is predicted using information provided by pixels in the previous bands in the same spatial position. As done in the JPEG-LS, the proposed coder also represents the mapped residuals by using an adaptive Golomb-Rice code with context modeling. This residual coding is context adaptive, where the context used for the current sample is identified by a context quantization function of the three gradients. Then, context-dependent Golomb-Rice code and bias parameters are estimated sample by sample. The proposed scheme was compared with three algorithms applied to the lossless compression of multispectral images, namely JPEG-LS, Rice coding, and JPEG2000. Simulation tests performed on AVIRIS images have demonstrated that the proposed compression scheme is suitable for multispectral images.

  17. Retinal oxygen saturation evaluation by multi-spectral fundus imaging

    NASA Astrophysics Data System (ADS)

    Khoobehi, Bahram; Ning, Jinfeng; Puissegur, Elise; Bordeaux, Kimberly; Balasubramanian, Madhusudhanan; Beach, James

    2007-03-01

    Purpose: To develop a multi-spectral method to measure oxygen saturation of the retina in the human eye. Methods: Five Cynomolgus monkeys with normal eyes were anesthetized with intramuscular ketamine/xylazine and intravenous pentobarbital. Multi-spectral fundus imaging was performed in five monkeys with a commercial fundus camera equipped with a liquid crystal tuned filter in the illumination light path and a 16-bit digital camera. Recording parameters were controlled with software written specifically for the application. Seven images at successively longer oxygen-sensing wavelengths were recorded within 4 seconds. Individual images for each wavelength were captured in less than 100 msec of flash illumination. Slightly misaligned images of separate wavelengths due to slight eye motion were registered and corrected by translational and rotational image registration prior to analysis. Numerical values of relative oxygen saturation of retinal arteries and veins and the underlying tissue in between the artery/vein pairs were evaluated by an algorithm previously described, but which is now corrected for blood volume from averaged pixels (n > 1000). Color saturation maps were constructed by applying the algorithm at each image pixel using a Matlab script. Results: Both the numerical values of relative oxygen saturation and the saturation maps correspond to the physiological condition, that is, in a normal retina, the artery is more saturated than the tissue and the tissue is more saturated than the vein. With the multi-spectral fundus camera and proper registration of the multi-wavelength images, we were able to determine oxygen saturation in the primate retinal structures on a tolerable time scale which is applicable to human subjects. Conclusions: Seven wavelength multi-spectral imagery can be used to measure oxygen saturation in retinal artery, vein, and tissue (microcirculation). This technique is safe and can be used to monitor oxygen uptake in humans. This work

  18. Mesospheric nightglow spectral survey taken by the ISO spectral spatial imager on ATLAS 1

    NASA Technical Reports Server (NTRS)

    Owens, J. K.; Torr, D. G.; Torr, M. R.; Chang, T.; Fennelly, J. A.; Richards, P. G.; Morgan, M. F.; Baldridge, T. W.; Fellows, C. W.; Dougani, H.

    1993-01-01

    This paper reports the first comprehensive spectral survey of the mesospheric airglow between 260 and 832 nm taken by the Imaging Spectrometric Observatory on the ATLAS 1 mission. We select data taken in the spectral window between 275 and 300 nm to determine the variation with altitude of the Herzberg I bands originating from the vibrational levels v-prime = 3 to 8. These data provide the first spatially resolved spectral measurements of the system. The data are used to demonstrate that to within an uncertainty of +/- 10 percent, the vibrational distribution remains invariant with altitude. The deficit reported previously for the v-prime = 5 level is not observed although there is a suggestion of depletion in v-prime = 6. The data could be used to place tight constraints on the vibrational dependence of quenching rate coefficients, and on the abundance of atomic oxygen.

  19. Mesospheric nightglow spectral survey taken by the ISO spectral spatial imager on Atlas 1

    NASA Technical Reports Server (NTRS)

    Owens, J. K.; Torr, D. G.; Torr, M. R.; Chang, T.; Fennelly, J. A.; Richards, P. G.; Morgan, M. F.; Baldridge, T. W.; Dougani, H.; Swift, W.

    1993-01-01

    This paper reports the first comprehensive spectral survey of the mesospheric airglow between 260 and 832 nm taken by the Imaging Spectrometric Observatory (ISO) on the ATLAS I mission. We select data taken in the spectral window between 275 and 300 nm to determine the variation with altitude of the Herzberg I bands originating from the vibrational levels v' = 3 to 8. These data provide the first spatially resolved spectral measurements of the system. The data are used to demonstrate that to within an uncertainty of + 10%, the vibrational distribution remains invariant with altitude. The deficit reported previously for the v' = 5 level is not observed although there is a suggestion of depletion in v' = 6. The data could be used to place tight constraints on the vibrational dependence of quenching rate coefficients, and on the abundance of atomic oxygen.

  20. Rapid microscopy measurement of very large spectral images.

    PubMed

    Lindner, Moshe; Shotan, Zav; Garini, Yuval

    2016-05-01

    The spectral content of a sample provides important information that cannot be detected by the human eye or by using an ordinary RGB camera. The spectrum is typically a fingerprint of the chemical compound, its environmental conditions, phase and geometry. Thus measuring the spectrum at each point of a sample is important for a large range of applications from art preservation through forensics to pathological analysis of a tissue section. To date, however, there is no system that can measure the spectral image of a large sample in a reasonable time. Here we present a novel method for scanning very large spectral images of microscopy samples even if they cannot be viewed in a single field of view of the camera. The system is based on capturing information while the sample is being scanned continuously 'on the fly'. Spectral separation implements Fourier spectroscopy by using an interferometer mounted along the optical axis. High spectral resolution of ~5 nm at 500 nm could be achieved with a diffraction-limited spatial resolution. The acquisition time is fairly high and takes 6-8 minutes for a sample size of 10mm x 10mm measured under a bright-field microscope using a 20X magnification. PMID:27137565

  1. Simulating Colour Vision Deficiency from a Spectral Image.

    PubMed

    Shrestha, Raju

    2016-01-01

    People with colour vision deficiency (CVD) have difficulty seeing full colour contrast and can miss some of the features in a scene. As a part of universal design, researcher have been working on how to modify and enhance the colour of images in order to make them see the scene with good contrast. For this, it is important to know how the original colour image is seen by different individuals with CVD. This paper proposes a methodology to simulate accurate colour deficient images from a spectral image using cone sensitivity of different cases of deficiency. As the method enables generation of accurate colour deficient image, the methodology is believed to help better understand the limitations of colour vision deficiency and that in turn leads to the design and development of more effective imaging technologies for better and wider accessibility in the context of universal design. PMID:27534332

  2. A pragmatic guide to multiphoton microscope design

    PubMed Central

    Young, Michael D.; Field, Jeffrey J.; Sheetz, Kraig E.; Bartels, Randy A.; Squier, Jeff

    2016-01-01

    Multiphoton microscopy has emerged as a ubiquitous tool for studying microscopic structure and function across a broad range of disciplines. As such, the intent of this paper is to present a comprehensive resource for the construction and performance evaluation of a multiphoton microscope that will be understandable to the broad range of scientific fields that presently exploit, or wish to begin exploiting, this powerful technology. With this in mind, we have developed a guide to aid in the design of a multiphoton microscope. We discuss source selection, optical management of dispersion, image-relay systems with scan optics, objective-lens selection, single-element light-collection theory, photon-counting detection, image rendering, and finally, an illustrated guide for building an example microscope. PMID:27182429

  3. RECENT PROGRESS IN MULTIFOCAL MULTIPHOTON MICROSCOPY

    PubMed Central

    LIU, LIXIN; SHAO, YONGHONG; NIU, HANBEN

    2013-01-01

    Multifocal multiphoton microscopy (MMM) has recently become an important tool in biomedicine for performing three-dimensional fast fluorescence imaging. Using various beamsplitting techniques, MMM splits the near-infrared laser beam into multiple beamlets and produces a multifocal array on the sample for parallel multiphoton excitation and then records fluorescence signal from all foci simultaneously with an area array detector, which significantly improves the imaging speed of multiphoton microscopy and allows for high efficiency in use of the excitation light. In this paper, we discuss the features of several MMM setups using different beamsplitting devices, including a Nipkow spinning disk, a microlens array, a set of beamsplitting mirrors, or a diffractive optical element (DOE). In particular, we present our recent work on the development of an MMM using a spatial light modulator (SLM). PMID:24363782

  4. Spectral imaging and biomedicine: new devices, new approaches

    SciTech Connect

    Levenson, Richard M.; Cronin, P. J.; Harvey, N. R.

    2002-01-01

    The advent of molecular medicine and new demands on pathologists to deliver prognostic and therapy-shaping analyses has created a need for enhanced imaging tools. Spectral imaging coupled wilh microscopy is a relatively novel and largely unexplored technology that holds out promise of satisfying, at least in part, such a need. New optical methods for spectral discrimination are being combined with powerful software approaches, often originally developed in diffbrent fields, to explore and exploit a wealth of informaticm beyond the capabilities of conventional color-based imaging approaches. Some of the new devices and software tools are described and illustrated here. While the results are indeed promising, it must be stressed that this field is in its infancy, and the optimal uses of this technology in the clinical arena still await definition.

  5. Acousto-optic infrared spectral imager for Pluto fast flyby

    NASA Technical Reports Server (NTRS)

    Glenar, D. A.; Hillman, J. J.

    1993-01-01

    Acousto-optic tunable filters (AOTF's) enable the design of compact, two-dimensional imaging spectrometers with high spectral and spatial resolution and with no moving parts. Tellurium dioxide AOTF's operate from about 400 nm to nearly 5 microns, and a single device will tune continuously over one octave by changing the RF acoustic frequency applied to the device. An infrared (1.2-2.5 micron) Acousto-Optic Imaging Spectrometer (AImS) was designed that closely conforms to the surface composition mapping objectives of the Pluto Fast Flyby. It features a 75-cm focal length telescope, infrared AOTF, and 256 x 256 NICMOS-3 focal plane array for acquiring narrowband images with a spectral resolving power (lambda/delta(lambda)) exceeding 250. We summarize the instrument design features and its expected performance at the Pluto-Charon encounter.

  6. Current developments in clinical multiphoton tomography

    NASA Astrophysics Data System (ADS)

    König, Karsten; Weinigel, Martin; Breunig, Hans Georg; Gregory, Axel; Fischer, Peter; Kellner-Höfer, Marcel; Bückle, Rainer

    2010-02-01

    Two-photon microscopy has been introduced in 1990 [1]. 13 years later, CE-marked clinical multiphoton systems for 3D imaging of human skin with subcellular resolution have been launched by the JenLab company with the tomograph DermaInspectTM. In 2010, the second generation of clinical multiphoton tomographs was introduced. The novel mobile multiphoton tomograph MPTflexTM, equipped with a flexible articulated optical arm, provides an increased flexibility and accessibility especially for clinical and cosmetical examinations. The multiphoton excitation of fluorescent biomolecules like NAD(P)H, flavins, porphyrins, elastin, and melanin as well as the second harmonic generation of collagen is induced by picojoule femtosecond laser pulses from an tunable turn-key near infrared laser system. The ability for rapid highquality image acquisition, the user-friendly operation of the system, and the compact and flexible design qualifies this system to be used for melanoma detection, diagnostics of dermatological disorders, cosmetic research, and skin aging measurements as well as in situ drug monitoring and animal research. So far, more than 1,000 patients and volunteers have been investigated with the multiphoton tomographs in Europe, Asia, and Australia.

  7. High spectral resolution imager for solar induced fluorescence observation

    NASA Astrophysics Data System (ADS)

    Barducci, A.; Guzzi, D.; Lastri, C.; Marcoionni, P.; Nardino, V.; Pippi, I.; Raimondi, V.; Sandri, P.

    2011-11-01

    The use of high-resolution imagers for determination of solar-induced fluorescence of natural bodies by observing the infilling of Fraunhofer lines has been frequently adopted as a tool for vegetation characterization. The option to perform those measurements from airborne platforms was addressed in the past. In-field observations gave evidence of the main requirements for an imaging spectrometer to be used for Sun-induced fluorescence measurements such as high spectral resolution and fine radiometric accuracy needed to resolve the shape of observed Fraunhofer lines with a high level of accuracy. In this paper, some solutions for the design of a high spectral resolution push-broom imaging spectrometer for Sun-induced fluorescence measurements are analysed. The main constraints for the optical design are a spectral resolution better than 0.01 nm and a wide field of view. Due to the fine instrumental spectral resolution, bidimensional focal plane arrays characterized by high quantum efficiency, low read-out noise, and high sensitivity are requested. The development of a lightweight instrument is a benefit for aerospace implementations of this technology. First results coming from laboratory measurements and optical simulations are presented and discussed taking into account their feasibility.

  8. Hyperspectral image classification by collaboration of spatial and spectral information

    NASA Astrophysics Data System (ADS)

    Yan, Yu-zhou; Zhao, Yongqiang; Xue, Hui-feng; Kou, Xiao-dong; Liu, Yuanzheng

    2009-07-01

    The classification of hyperspectral image data has drawn much attention in recent years. Consequently, it contains not only spectral information of objects, but also spatial arrangement of objects. The most established Hyperspectral classifiers are based on the observed spectral signal, and ignore the spatial relations among observations. Information captured in neighboring locations may provide useful supplementary knowledge for analysis. To combine the spectral and spatial information in the classification process, in this paper, a Multidimensional Local Spatial Autocorrelation (MLSA) is proposed for hyperspectral image data. Based on this measure, a collaborative classification method is proposed, which integrates the spectral and spatial autocorrelation during the decision-making process. The trials of our experiment are conducted on two scenes, one from HYDICE 210-band imagery collected over an area that contains a diverse range of terrain features and the other is toy car hyperspectral image captured at Instrumentation and Sensing Laboratory (ISL) at Beltsville Agricultural Research Center. Quantitative measures of local consistency (smoothness) and global labeling, along with class maps, demonstrate the benefits of applying this method for unsupervised and supervised classification.

  9. Multiphoton imaging reveals that nanosecond pulsed electric fields collapse tumor and normal vascular perfusion in human glioblastoma xenografts

    PubMed Central

    Bardet, Sylvia M.; Carr, Lynn; Soueid, Malak; Arnaud-Cormos, Delia; Leveque, Philippe; O’Connor, Rodney P.

    2016-01-01

    Despite the biomedical advances of the last century, many cancers including glioblastoma are still resistant to existing therapies leaving patients with poor prognoses. Nanosecond pulsed electric fields (nsPEF) are a promising technology for the treatment of cancer that have thus far been evaluated in vitro and in superficial malignancies. In this paper, we develop a tumor organoid model of glioblastoma and apply intravital multiphoton microscopy to assess their response to nsPEFs. We demonstrate for the first time that a single 10 ns, high voltage electric pulse (35–45 kV/cm), collapses the perfusion of neovasculature, and also alters the diameter of capillaries and larger vessels in normal tissue. These results contribute to the fundamental understanding of nsPEF effects in complex tissue environments, and confirm the potential of nsPEFs to disrupt the microenvironment of solid tumors such as glioblastoma. PMID:27698479

  10. Wavelet based hyperspectral image restoration using spatial and spectral penalties

    NASA Astrophysics Data System (ADS)

    Rasti, Behnood; Sveinsson, Johannes R.; Ulfarsson, Magnus O.; Benediktsson, Jon A.

    2013-10-01

    In this paper a penalized least squares cost function with a new spatial-spectral penalty is proposed for hyper- spectral image restoration. The new penalty is a combination of a Group LASSO (GLASSO) and First Order Roughness Penalty (FORP) in the wavelet domain. The restoration criterion is solved using the Alternative Direction Method of Multipliers (ADMM). The results are compared with other restoration methods where the proposed method outperforms them for the simulated noisy data set based on Signal to Noise Ratio (SNR) and visually outperforms them on a real degraded data set.

  11. Online Unmixing of Multitemporal Hyperspectral Images Accounting for Spectral Variability.

    PubMed

    Thouvenin, Pierre-Antoine; Dobigeon, Nicolas; Tourneret, Jean-Yves

    2016-09-01

    Hyperspectral unmixing is aimed at identifying the reference spectral signatures composing a hyperspectral image and their relative abundance fractions in each pixel. In practice, the identified signatures may vary spectrally from an image to another due to varying acquisition conditions, thus inducing possibly significant estimation errors. Against this background, the hyperspectral unmixing of several images acquired over the same area is of considerable interest. Indeed, such an analysis enables the endmembers of the scene to be tracked and the corresponding endmember variability to be characterized. Sequential endmember estimation from a set of hyperspectral images is expected to provide improved performance when compared with methods analyzing the images independently. However, the significant size of the hyperspectral data precludes the use of batch procedures to jointly estimate the mixture parameters of a sequence of hyperspectral images. Provided that each elementary component is present in at least one image of the sequence, we propose to perform an online hyperspectral unmixing accounting for temporal endmember variability. The online hyperspectral unmixing is formulated as a two-stage stochastic program, which can be solved using a stochastic approximation. The performance of the proposed method is evaluated on synthetic and real data. Finally, a comparison with independent unmixing algorithms illustrates the interest of the proposed strategy.

  12. Online Unmixing of Multitemporal Hyperspectral Images Accounting for Spectral Variability.

    PubMed

    Thouvenin, Pierre-Antoine; Dobigeon, Nicolas; Tourneret, Jean-Yves

    2016-09-01

    Hyperspectral unmixing is aimed at identifying the reference spectral signatures composing a hyperspectral image and their relative abundance fractions in each pixel. In practice, the identified signatures may vary spectrally from an image to another due to varying acquisition conditions, thus inducing possibly significant estimation errors. Against this background, the hyperspectral unmixing of several images acquired over the same area is of considerable interest. Indeed, such an analysis enables the endmembers of the scene to be tracked and the corresponding endmember variability to be characterized. Sequential endmember estimation from a set of hyperspectral images is expected to provide improved performance when compared with methods analyzing the images independently. However, the significant size of the hyperspectral data precludes the use of batch procedures to jointly estimate the mixture parameters of a sequence of hyperspectral images. Provided that each elementary component is present in at least one image of the sequence, we propose to perform an online hyperspectral unmixing accounting for temporal endmember variability. The online hyperspectral unmixing is formulated as a two-stage stochastic program, which can be solved using a stochastic approximation. The performance of the proposed method is evaluated on synthetic and real data. Finally, a comparison with independent unmixing algorithms illustrates the interest of the proposed strategy. PMID:27305679

  13. Online Unmixing of Multitemporal Hyperspectral Images Accounting for Spectral Variability

    NASA Astrophysics Data System (ADS)

    Thouvenin, Pierre-Antoine; Dobigeon, Nicolas; Tourneret, Jean-Yves

    2016-09-01

    Hyperspectral unmixing is aimed at identifying the reference spectral signatures composing an hyperspectral image and their relative abundance fractions in each pixel. In practice, the identified signatures may vary spectrally from an image to another due to varying acquisition conditions, thus inducing possibly significant estimation errors. Against this background, hyperspectral unmixing of several images acquired over the same area is of considerable interest. Indeed, such an analysis enables the endmembers of the scene to be tracked and the corresponding endmember variability to be characterized. Sequential endmember estimation from a set of hyperspectral images is expected to provide improved performance when compared to methods analyzing the images independently. However, the significant size of hyperspectral data precludes the use of batch procedures to jointly estimate the mixture parameters of a sequence of hyperspectral images. Provided that each elementary component is present in at least one image of the sequence, we propose to perform an online hyperspectral unmixing accounting for temporal endmember variability. The online hyperspectral unmixing is formulated as a two-stage stochastic program, which can be solved using a stochastic approximation. The performance of the proposed method is evaluated on synthetic and real data. A comparison with independent unmixing algorithms finally illustrates the interest of the proposed strategy.

  14. Interpretation of AIS Images of Cuprite, Nevada Using Constraints of Spectral Mixtures

    NASA Technical Reports Server (NTRS)

    Smith, M. O.; Adams, J. B.

    1985-01-01

    A technique is outlined that tests the hypothesis Airborne Imaging Spectrometer (AIS) image spectra are produced by mixtures of surface materials. This technique allows separation of AIS images into concentration images of spectral endmembers (e.g., surface materials causing spectral variation). Using a spectral reference library it was possible to uniquely identify these spectral endmembers with respect to the reference library and to calibrate the AIS images.

  15. Reflectance and Fluorescence Spectral Recovery via Actively Lit RGB Images.

    PubMed

    Fu, Ying; Lam, Antony; Sato, Imari; Okabe, Takahiro; Sato, Yoichi

    2016-07-01

    In recent years, fluorescence analysis of scenes has received attention in computer vision. Fluorescence can provide additional information about scenes, and has been used in applications such as camera spectral sensitivity estimation, 3D reconstruction, and color relighting. In particular, hyperspectral images of reflective-fluorescent scenes provide a rich amount of data. However, due to the complex nature of fluorescence, hyperspectral imaging methods rely on specialized equipment such as hyperspectral cameras and specialized illuminants. In this paper, we propose a more practical approach to hyperspectral imaging of reflective-fluorescent scenes using only a conventional RGB camera and varied colored illuminants. The key idea of our approach is to exploit a unique property of fluorescence: the chromaticity of fluorescent emissions are invariant under different illuminants. This allows us to robustly estimate spectral reflectance and fluorescent emission chromaticity. We then show that given the spectral reflectance and fluorescent chromaticity, the fluorescence absorption and emission spectra can also be estimated. We demonstrate in results that all scene spectra can be accurately estimated from RGB images. Finally, we show that our method can be used to accurately relight scenes under novel lighting. PMID:27295456

  16. Quantitative study on appearance of microvessels in spectral endoscopic imaging

    NASA Astrophysics Data System (ADS)

    Yamaguchi, Hiroshi; Saito, Takaaki; Shiraishi, Yasushi; Arai, Fumihito; Morimoto, Yoshinori; Yuasa, Atsuko

    2015-03-01

    Increase in abnormal microvessels in the superficial mucosa is often relevant to diagnostic findings of neoplasia in digestive endoscopy; hence, observation of superficial vasculature is crucial for cancer diagnosis. To enhance the appearance of such vessels, several spectral endoscopic imaging techniques have been developed, such as narrow-band imaging and blue laser imaging. Both techniques exploit narrow-band blue light for the enhancement. The emergence of such spectral imaging techniques has increased the importance of understanding the relation of the light wavelength to the appearance of superficial vasculature, and thus a new method is desired for quantitative analysis of vessel visibility in relation to the actual structure in the tissue. Here, we developed microvessel-simulating phantoms that allowed quantitative evaluation of the appearance of 15-μm-thick vessels. We investigated the relation between the vascular contrast and light wavelength by the phantom measurements and also verified it in experiments with swine, where the endoscopically observed vascular contrast was investigated together with its real vascular depth and diameter obtained by microscopic observation of fluorescence-labeled vessels. Our study indicates that changing the spectral property even in the wavelength range of blue light may allow selective enhancement of the vascular depth for clinical use.

  17. ISIS; An Information-efficient Spectral Imaging System

    SciTech Connect

    Boye, C.A.; Descour, M.R.; Gentry, S.M.; Grotbeck, C.L.; Stallard, B.R.; Sweatt, W.C.

    1998-07-19

    A specialized hyperspectral imager has been developed that preprocesses the spectra from an image before the light reaches the detectors. This "optical computer" does not allow the flexibility of digital post-processing. However, the processing is done in real time and the system can examine = 2 x 10{sup 6} scene pixels/sec. Therefore, outdoors it could search for pollutants, vegetation types, minerals, or man-made objects. On a high- speed production line it could identify defects in sheet products like plastic wrap or film, or on painted or plastic parts. ISIS is a line scan imager. A spectrally dispersed slit image is projected on a Spatial Light Modulator. The SLM is programmed to take the inner product of the spectral intensity vector and a spectral basis vector. The SLM directs the positive and negative parts of the inner product to different linear detector arrays so the signal difference equals the inner product. We envision a system with one telescope and =4 SLMS.

  18. Changes in the redox state and endogenous fluorescence of in vivo human skin due to intrinsic and photo-aging, measured by multiphoton tomography with fluorescence lifetime imaging

    NASA Astrophysics Data System (ADS)

    Sanchez, Washington Y.; Obispo, Clara; Ryan, Elizabeth; Grice, Jeffrey E.; Roberts, Michael S.

    2013-06-01

    Ultraviolet radiation from solar exposure is a key extrinsic factor responsible for premature skin aging (i.e., photo-aging). Recent advances using in vivo multiphoton tomography (MPT) demonstrate the efficacy of this approach to assess intrinsic and extrinsic skin aging as an alternative to existing invasive techniques. In this study, we measured changes in epidermal autofluorescence, dermal collagen second harmonic generation (SHG), and the redox state of solar-exposed and solar-protected human skin by MPT with fluorescence lifetime imaging (MPT-FLIM). Twenty-four volunteers across four age categories (20 to 29, 30 to 39, 40 to 49, and 50 to 59 years old; six volunteers each) were recruited for MPT-FLIM imaging of the dorsal (solar-exposed; photo-damaged) and volar (solar-protected) forearm. We demonstrate a higher intensity of dermal collagen SHG within the volar forearm compared to dorsal solar-exposed skin. Redox imaging of each epidermal skin stratum by FLIM demonstrates an increase in fluorescence lifetime in the solar-exposed dorsal forearm that is more apparent in aged skin. The results of this study suggest the redox state of the viable epidermis is a key marker in assessing intrinsic and photo-damage skin aging, in combination with changes in autofluorescence and SHG.

  19. LED-based endoscopic light source for spectral imaging

    NASA Astrophysics Data System (ADS)

    Browning, Craig M.; Mayes, Samuel; Favreau, Peter; Rich, Thomas C.; Leavesley, Silas J.

    2016-03-01

    Colorectal cancer is the United States 3rd leading cancer in death rates.1 The current screening for colorectal cancer is an endoscopic procedure using white light endoscopy (WLE). There are multiple new methods testing to replace WLE, for example narrow band imaging and autofluorescence imaging.2 However, these methods do not meet the need for a higher specificity or sensitivity. The goal for this project is to modify the presently used endoscope light source to house 16 narrow wavelength LEDs for spectral imaging in real time while increasing sensitivity and specificity. The process to do such was to take an Olympus CLK-4 light source, replace the light and electronics with 16 LEDs and new circuitry. This allows control of the power and intensity of the LEDs. This required a larger enclosure to house a bracket system for the solid light guide (lightpipe), three new circuit boards, a power source and National Instruments hardware/software for computer control. The results were a successfully designed retrofit with all the new features. The LED testing resulted in the ability to control each wavelength's intensity. The measured intensity over the voltage range will provide the information needed to couple the camera for imaging. Overall the project was successful; the modifications to the light source added the controllable LEDs. This brings the research one step closer to the main goal of spectral imaging for early detection of colorectal cancer. Future goals will be to connect the camera and test the imaging process.

  20. Images and Spectral Performance of WFC3 Interference Filters

    NASA Technical Reports Server (NTRS)

    Quijada, Manuel A.; Boucarut, R.; Telfer, R.; Baggett, S.; Quijano, J. Kim; Allen, George; Arsenovic, Peter

    2006-01-01

    The Wide Field Camera 3 (WFC3) is a panchromatic imager that will be deployed in the Hubble Space Telescope (HST). The mission of the WFC3 is to enhance HST1s imaging capability in the ultraviolet, visible and near-infrared spectral regions. Together with a wavelength coverage spanning 2000A to 1.7 micron, the WFC3 high sensitivity, high spatial resolution, and large field-of-view provide the astronomer with an unprecedented set of tools for exploring all types of exciting astrophysical terrain and for addressing many key questions in astronomy today. The filter compliment, which includes broad, medium, and narrow band filters, naturally reflects the diversity of astronomical programs to be targeted with WFC3. The WFC3 holds 61 UVIS filters elements, 14 IR filters, and 3 dispersive elements. During ground testing, the majority of the UVIS filters were found to exhibit excellent performance consistent with or exceeding expectations; however, a subset of filters showed considerable ghost images; some with relative intensity as high as 10-15%. Replacement filters with band-defining coatings that substantially reduce these ghost images were designed and procured. A state-of-the-art characterization setup was developed to measured the intensity of ghost images, focal shift, wedge direction , transmitted uniformity and surface feature of filters that could effect uniform flat field images. We will report on this new filter characterization methods, as well as the spectral performance measurements of the in-band transmittance and blocking.

  1. Hybrid Image Fusion for Sharpness Enhancement of Multi-Spectral Lunar Images

    NASA Astrophysics Data System (ADS)

    Awumah, Anna; Mahanti, Prasun; Robinson, Mark

    2016-10-01

    Image fusion enhances the sharpness of a multi-spectral (MS) image by incorporating spatial details from a higher-resolution panchromatic (Pan) image [1,2]. Known applications of image fusion for planetary images are rare, although image fusion is well-known for its applications to Earth-based remote sensing. In a recent work [3], six different image fusion algorithms were implemented and their performances were verified with images from the Lunar Reconnaissance Orbiter (LRO) Camera. The image fusion procedure obtained a high-resolution multi-spectral (HRMS) product from the LRO Narrow Angle Camera (used as Pan) and LRO Wide Angle Camera (used as MS) images. The results showed that the Intensity-Hue-Saturation (IHS) algorithm results in a high-spatial quality product while the Wavelet-based image fusion algorithm best preserves spectral quality among all the algorithms. In this work we show the results of a hybrid IHS-Wavelet image fusion algorithm when applied to LROC MS images. The hybrid method provides the best HRMS product - both in terms of spatial resolution and preservation of spectral details. Results from hybrid image fusion can enable new science and increase the science return from existing LROC images.[1] Pohl, Cle, and John L. Van Genderen. "Review article multisensor image fusion in remote sensing: concepts, methods and applications." International journal of remote sensing 19.5 (1998): 823-854.[2] Zhang, Yun. "Understanding image fusion." Photogramm. Eng. Remote Sens 70.6 (2004): 657-661.[3] Mahanti, Prasun et al. "Enhancement of spatial resolution of the LROC Wide Angle Camera images." Archives, XXIII ISPRS Congress Archives (2016).

  2. Solar abundances with the SPICE spectral imager on Solar Orbiter

    NASA Astrophysics Data System (ADS)

    Giunta, Alessandra; Haberreiter, Margit; Peter, Hardi; Vial, Jean-Claude; Harrison, Richard; Parenti, Susanna; Innes, Davina; Schmutz, Werner; Buchlin, Eric; Chamberlin, Phillip; Thompson, William; Bocchialini, Karine; Gabriel, Alan; Morris, Nigel; Caldwell, Martin; Auchere, Frederic; Curdt, Werner; Teriaca, Luca; Hassler, Donald M.; DeForest, Craig; Hansteen, Viggo; Carlsson, Mats; Philippon, Anne; Janvier, Miho; Wimmer-Schweingruber, Robert; Griffin, Douglas; Baudin, Frederic; Davila, Joseph; Fludra, Andrzej; Waltham, Nick; Eccleston, Paul; Gottwald, Alexander; Klein, Roman; Hanley, John; Walls, Buddy; Howe, Chris; Schuehle, Udo; Gyo, Manfred; Pfiffner, Dany

    2016-07-01

    Elemental composition of the solar atmosphere and in particular abundance bias of low and high First Ionization Potential (FIP) elements are a key tracer of the source regions of the solar wind. These abundances and their spatio-temporal variations, as well as the other plasma parameters , will be derived by the SPICE (Spectral Imaging of the Coronal Environment) EUV spectral imager on the upcoming Solar Orbiter mission. SPICE is designed to provide spectroheliograms (spectral images) using a core set of emission lines arising from ions of both low-FIP and high-FIP elements. These lines are formed over a wide range of temperatures, enabling the analysis of the different layers of the solar atmosphere. SPICE will use these spectroheliograms to produce dynamic composition maps of the solar atmosphere to be compared to in-situ measurements of the solar wind composition of the same elements (i.e. O, Ne, Mg, Fe). This will provide a tool to study the connectivity between the spacecraft (the Heliosphere) and the Sun. We will discuss the SPICE capabilities for such composition measurements.

  3. Single-shot ultrafast tomographic imaging by spectral multiplexing

    NASA Astrophysics Data System (ADS)

    Matlis, N. H.; Axley, A.; Leemans, W. P.

    2012-10-01

    Computed tomography has profoundly impacted science, medicine and technology by using projection measurements scanned over multiple angles to permit cross-sectional imaging of an object. The application of computed tomography to moving or dynamically varying objects, however, has been limited by the temporal resolution of the technique, which is set by the time required to complete the scan. For objects that vary on ultrafast timescales, traditional scanning methods are not an option. Here we present a non-scanning method capable of resolving structure on femtosecond timescales by using spectral multiplexing of a single laser beam to perform tomographic imaging over a continuous range of angles simultaneously. We use this technique to demonstrate the first single-shot ultrafast computed tomography reconstructions and obtain previously inaccessible structure and position information for laser-induced plasma filaments. This development enables real-time tomographic imaging for ultrafast science, and offers a potential solution to the challenging problem of imaging through scattering surfaces.

  4. Multiphoton ionization spectra of radical products in the F(2P)+ketene system: Spectral assignments and formation reaction for CH2F, observation of CF and CH

    NASA Astrophysics Data System (ADS)

    Hudgens, Jeffrey W.; Dulcey, C. S.; Long, George R.; Bogan, Denis J.

    1987-10-01

    The reactions of F(2P)+ketene and F(2P)+ketene-d2 were studied in a flow reactor. Spectra of the radical products CH2F, CD2F, CH, CF, and atomic carbon were detected between 292-395 nm by resonance enhanced multiphoton ionization (REMPI) mass spectrometry. Fluoromethyl radicals were identified as a major product of the F+ketene elementary reaction. REMPI spectra of fluoromethyl radicals originated from two-photon preparation of 3p, 4p, and 5p Rydberg states (quantum defect ˜0.6). Absorption of a third laser photon ionized the radicals, a 2+1 REMPI mechanism. Rydberg band origins were observed in CH2F at ν00=52 863, 63 275, and 67 265 cm-1 and in CD2F at ν00=52 786, 63 195, and 67 186 cm-1. A normal mode analysis revealed the vibrational frequencies of the C-F stretch, CH2 scissors, and out-of-plane bending modes in the Rydberg states. The ground state out-of-plane bending frequency in CH2F is 260(30) cm-1 and in CD2F it is 170(30) cm-1. CH radicals were generated by the photolysis of ketene and observed at ˜311 nm by two-photon excitation through the D 2Π(v'=2) ←←X 2Πr band. The reaction mechanism that generated the CF radicals was not determined. The REMPI CF radical spectrum generated by a 2+1 photon mechanism appeared as a series of bandheads described by the constants ν00=˜52 572 cm-1, ωe =1820 cm-1, and ωexe =-18.2 cm-1.

  5. Biomass estimator for NIR image with a few additional spectral band images taken from light UAS

    NASA Astrophysics Data System (ADS)

    Pölönen, Ilkka; Salo, Heikki; Saari, Heikki; Kaivosoja, Jere; Pesonen, Liisa; Honkavaara, Eija

    2012-05-01

    A novel way to produce biomass estimation will offer possibilities for precision farming. Fertilizer prediction maps can be made based on accurate biomass estimation generated by a novel biomass estimator. By using this knowledge, a variable rate amount of fertilizers can be applied during the growing season. The innovation consists of light UAS, a high spatial resolution camera, and VTT's novel spectral camera. A few properly selected spectral wavelengths with NIR images and point clouds extracted by automatic image matching have been used in the estimation. The spectral wavelengths were chosen from green, red, and NIR channels.

  6. Differential Multiphoton Laser Scanning Microscopy

    PubMed Central

    Field, Jeffrey J.; Sheetz, Kraig E.; Chandler, Eric V.; Hoover, Erich E.; Young, Michael D.; Ding, Shi-you; Sylvester, Anne W.; Kleinfeld, David; Squier, Jeff A.

    2016-01-01

    Multifocal multiphoton microscopy (MMM) in the biological and medical sciences has become an important tool for obtaining high resolution images at video rates. While current implementations of MMM achieve very high frame rates, they are limited in their applicability to essentially those biological samples that exhibit little or no scattering. In this paper, we report on a method for MMM in which imaging detection is not necessary (single element point detection is implemented), and is therefore fully compatible for use in imaging through scattering media. Further, we demonstrate that this method leads to a new type of MMM wherein it is possible to simultaneously obtain multiple images and view differences in excitation parameters in a single shot. PMID:27390511

  7. Evaluation of the oxidative stress of psoriatic fibroblasts based on spectral two-photon fluorescence lifetime imaging

    NASA Astrophysics Data System (ADS)

    Kapsokalyvas, Dimitrios; Barygina, Victoria; Cicchi, Riccardo; Fiorillo, Claudia; Pavone, Francesco S.

    2013-02-01

    Psoriasis is an autoimmune disease of the skin characterized by hyperkeratosis, hyperproliferation of the epidermis, inflammatory cell accumulation and increased dilatation of dermal papillary blood vessels. Metabolic activity is increased in the epidermis and the dermis. Oxidative stress is high mainly due to reactive oxygen species (ROS) originating from the skin environment and cellular metabolism. We employed a custom multiphoton microscope coupled with a FLIM setup to image primary culture fibroblast cells from perilesional and lesional psoriatic skin in-vitro. Twophoton excited fluorescence images revealed the morphological differences between healthy and psoriatic fibroblasts. Based on the spectral analysis of the NADH and FAD components the oxidative stress was assessed and found to be higher in psoriatic cells. Furthermore the fluorescence lifetime properties were investigated with a TCSPC FLIM module. Mean fluorescence lifetime was found to be longer in psoriatic lesional cells. Analysis of the fast (τ1) and slow (τ2) decay lifetimes revealed a decrease of the ratio of the contribution of the fast (α1) parameter to the contribution of the slow (α2) parameter. The fluorescence in the examined part of the spectrum is attributed mainly to NADH. The decrease of the ratio (α1)/ (α2) is believed to correlate strongly with the anti-oxidant properties of NADH which can lead to the variation of its population in high ROS environment. This methodology could serve as an index of the oxidative status in cells and furthermore could be used to probe the oxidative stress of tissues in-vivo.

  8. Thermal Infrared Spectral Imager for Airborne Science Applications

    NASA Technical Reports Server (NTRS)

    Johnson, William R.; Hook, Simon J.; Mouroulis, Pantazis; Wilson, Daniel W.; Gunapala, Sarath D.; Hill, Cory J.; Mumolo, Jason M.; Eng, Bjorn T.

    2009-01-01

    An airborne thermal hyperspectral imager is under development which utilizes the compact Dyson optical configuration and quantum well infrared photo detector (QWIP) focal plane array. The Dyson configuration uses a single monolithic prism-like grating design which allows for a high throughput instrument (F/1.6) with minimal ghosting, stray-light and large swath width. The configuration has the potential to be the optimal imaging spectroscopy solution for lighter-than-air (LTA) vehicles and unmanned aerial vehicles (UAV) due to its small form factor and relatively low power requirements. The planned instrument specifications are discussed as well as design trade-offs. Calibration testing results (noise equivalent temperature difference, spectral linearity and spectral bandwidth) and laboratory emissivity plots from samples are shown using an operational testbed unit which has similar specifications as the final airborne system. Field testing of the testbed unit was performed to acquire plots of apparent emissivity for various known standard minerals (such as quartz). A comparison is made using data from the ASTER spectral library.

  9. Multiphoton microscopy and microspectroscopy for diagnostics of inflammatory and neoplastic lung

    NASA Astrophysics Data System (ADS)

    Pavlova, Ina; Hume, Kelly R.; Yazinski, Stephanie A.; Flanders, James; Southard, Teresa L.; Weiss, Robert S.; Webb, Watt W.

    2012-03-01

    Limitations of current medical procedures for detecting early lung cancers inspire the need for new diagnostic imaging modalities for the direct microscopic visualization of lung nodules. Multiphoton microscopy (MPM) provides for subcellular resolution imaging of intrinsic fluorescence from unprocessed tissue with minimal optical attenuation and photodamage. We demonstrate that MPM detects morphological and spectral features of lung tissue and differentiates between normal, inflammatory and neoplastic lung. Ex vivo MPM imaging of intrinsic two-photon excited fluorescence was performed on mouse and canine neoplastic, inflammatory and tumor-free lung sites. Results showed that MPM detected microanatomical differences between tumor-free and neoplastic lung tissue similar to standard histopathology but without the need for tissue processing. Furthermore, inflammatory sites displayed a distinct red-shifted fluorescence compared to neoplasms in both mouse and canine lung, and adenocarcinomas displayed a less pronounced fluorescence emission in the 500 to 550 nm region compared to adenomas in mouse models of lung cancer. These spectral distinctions were also confirmed by two-photon excited fluorescence microspectroscopy. We demonstrate the feasibility of applying MPM imaging of intrinsic fluorescence for the differentiation of lung neoplasms, inflammatory and tumor-free lung, which motivates the application of multiphoton endoscopy for the in situ imaging of lung nodules.

  10. An Integrated Imaging Detector of Polarization and Spectral Content

    NASA Technical Reports Server (NTRS)

    Rust, D. M.; Thompson, K. E.

    1993-01-01

    A new type of image detector has been designed to simultaneously analyze the polarization of light at all picture elements in a scene. The Integrated Dual Imaging Detector (IDID) consists of a polarizing beamsplitter bonded to a charge-coupled device (CCD), with signal-analysis circuitry and analog-to-digital converters, all integrated on a silicon chip. It should be capable of 1:10(exp 4) polarization discrimination. The IDID should simplify the design and operation of imaging polarimeters and spectroscopic imagers used, for example, in atmospheric and solar research. Innovations in the IDID include (1) two interleaved 512 x 1024-pixel imaging arrays (one for each polarization plane); (2) large dynamic range (well depth of 10(exp 6) electrons per pixel); (3) simultaneous readout of both images at 10 million pixels per second each; (4) on-chip analog signal processing to produce polarization maps in real time; (5) on-chip 10-bit A/D conversion. When used with a lithium-niobate Fabry-Perot etalon or other color filter that can encode spectral information as polarization, the IDID can collect and analyze simultaneous images at two wavelengths. Precise photometric analysis of molecular or atomic concentrations in the atmosphere is one suggested application. When used in a solar telescope, the IDID will charge the polarization, which can then be converted to maps of the vector magnetic fields on the solar surface.

  11. Modeling and image reconstruction in spectrally resolved bioluminescence tomography

    NASA Astrophysics Data System (ADS)

    Dehghani, Hamid; Pogue, Brian W.; Davis, Scott C.; Patterson, Michael S.

    2007-02-01

    Recent interest in modeling and reconstruction algorithms for Bioluminescence Tomography (BLT) has increased and led to the general consensus that non-spectrally resolved intensity-based BLT results in a non-unique problem. However, the light emitted from, for example firefly Luciferase, is widely distributed over the band of wavelengths from 500 nm to 650 nm and above, with the dominant fraction emitted from tissue being above 550 nm. This paper demonstrates the development of an algorithm used for multi-wavelength 3D spectrally resolved BLT image reconstruction in a mouse model. It is shown that using a single view data, bioluminescence sources of up to 15 mm deep can be successfully recovered given correct information about the underlying tissue absorption and scatter.

  12. In vivo simultaneous multispectral fluorescence imaging with spectral multiplexed volume holographic imaging system

    NASA Astrophysics Data System (ADS)

    Lv, Yanlu; Zhang, Jiulou; Zhang, Dong; Cai, Wenjuan; Chen, Nanguang; Luo, Jianwen

    2016-06-01

    A simultaneous multispectral fluorescence imaging system incorporating multiplexed volume holographic grating (VHG) is developed to acquire multispectral images of an object in one shot. With the multiplexed VHG, the imaging system can provide the distribution and spectral characteristics of multiple fluorophores in the scene. The implementation and performance of the simultaneous multispectral imaging system are presented. Further, the system's capability in simultaneously obtaining multispectral fluorescence measurements is demonstrated with in vivo experiments on a mouse. The demonstrated imaging system has the potential to obtain multispectral images fluorescence simultaneously.

  13. Hindlimb heating increases vascular access of large molecules to murine tibial growth plates measured by in vivo multiphoton imaging

    PubMed Central

    Efaw, Morgan L.; Williams, Rebecca M.

    2013-01-01

    Advances in understanding the molecular regulation of longitudinal growth have led to development of novel drug therapies for growth plate disorders. Despite progress, a major unmet challenge is delivering therapeutic agents to avascular-cartilage plates. Dense extracellular matrix and lack of penetrating blood vessels create a semipermeable “barrier,” which hinders molecular transport at the vascular-cartilage interface. To overcome this obstacle, we used a hindlimb heating model to manipulate bone circulation in 5-wk-old female mice (n = 22). Temperatures represented a physiological range of normal human knee joints. We used in vivo multiphoton microscopy to quantify temperature-enhanced delivery of large molecules into tibial growth plates. We tested the hypothesis that increasing hindlimb temperature from 22°C to 34°C increases vascular access of large systemic molecules, modeled using 10, 40, and 70 kDa dextrans that approximate sizes of physiological regulators. Vascular access was quantified by vessel diameter, velocity, and dextran leakage from subperichondrial plexus vessels and accumulation in growth plate cartilage. Growth plate entry of 10 kDa dextrans increased >150% at 34°C. Entry of 40 and 70 kDa dextrans increased <50%, suggesting a size-dependent temperature enhancement. Total dextran levels in the plexus increased at 34°C, but relative leakage out of vessels was not temperature dependent. Blood velocity and vessel diameter increased 118% and 31%, respectively, at 34°C. These results demonstrate that heat enhances vascular carrying capacity and bioavailability of large molecules around growth plates, suggesting that temperature could be a noninvasive strategy for modulating delivery of therapeutics to impaired growth plates of children. PMID:24371019

  14. Dual-order snapshot spectral imaging of plasmonic nanoparticles

    NASA Astrophysics Data System (ADS)

    Nusz, Gregory J.; Marinakos, Stella M.; Rangarajan, Srinath; Chilkoti, Ashutosh

    2011-07-01

    The development of truly scalable, multiplexed optical microarrays requires a detection platform capable of simultaneous detection of multiple signals in real-time. We present a technique we term dual-order snapshot spectroscopic imaging (DOSSI) and demonstrate that it can be effectively used to collect spectrally resolved images of a full field of view of sparsely located spots in real time. Resonant peaks of plasmonic gold nanoparticles were tracked as a function of their surrounding refractive index. Measurement uncertainty analysis indicated that the spectral resolution of DOSSI in the described configuration is approximately 0.95nm. Further, real-time measurements by DOSSI allowed discrimination between optically identical nanoparticles that were functionalized with two homologous small molecule ligands that bound to the same protein, albeit with different affinity, based purely on their different molecular interaction kinetics---a feat not possible with slower raster-type hyperspectral imaging systems, or other dark-field optical detection systems that solely rely on end point measurements. Kinetic measurements of plasmon bands by DOSSI can be performed with a relatively simple optical system, thereby opening up the possibility of developing low-cost detectors for arrayed plasmonic diagnostics.

  15. Singular value decomposition based regularization prior to spectral mixing improves crosstalk in dynamic imaging using spectral diffuse optical tomography.

    PubMed

    Zhan, Yuxuan; Eggebrecht, Adam T; Culver, Joseph P; Dehghani, Hamid

    2012-09-01

    The spectrally constrained diffuse optical tomography (DOT) method relies on incorporating spectral prior information directly into the image reconstruction algorithm, thereby correlating the underlying optical properties across multiple wavelengths. Although this method has been shown to provide a solution that is stable, the use of conventional Tikhonov-type regularization techniques can lead to additional crosstalk between parameters, particularly in linear, single-step dynamic imaging applications. This is due mainly to the suboptimal regularization of the spectral Jacobian matrix, which smoothes not only the image-data space, but also the spectral mapping space. In this work a novel regularization technique based on the singular value decomposition (SVD) is presented that preserves the spectral prior information while regularizing the Jacobian matrix, leading to dramatically reduced crosstalk between the recovered parameters. Using simulated data, images of changes in oxygenated and deoxygenated hemoglobin concentrations are reconstructed via the SVD-based approach and compared with images reconstructed by using non-spectral and conventional spectral methods. In a 2D, two wavelength example, it is shown that the proposed approach provides a 98% reduction in crosstalk between recovered parameters as compared with conventional spectral reconstruction algorithms, and 60% as compared with non-spectrally constrained algorithms. Using a subject specific multilayered model of the human head, a noiseless dynamic simulation of cortical activation is performed to further demonstrate such improvement in crosstalk. However, with the addition of realistic noise in the data, both non-spectral and proposed algorithms perform similarly, indicating that the use of spectrally constrained reconstruction algorithms in dynamic DOT may be limited by the contrast of the signal as well as the noise characteristics of the system.

  16. Multiphoton electromagnetically induced transparency

    NASA Astrophysics Data System (ADS)

    Wen, Lingling; Kang, Hoonsoo; Zhu, Yifu; Wu, Ying

    2003-05-01

    We show that in multi-level atomic systems coupled by multiple laser fields, all linear and nonlinear absorptions may be completely suppressed, leading to the multiphoton electromagnetically induced transparency (EIT). Under suitable conditions, multiphoton EIT may be used to realize selective steady-state population inversion in coherently pumped atomic systems and achieve efficient nonlinear light generation at low light intensities. As examples, we will present studies of multiphoton EIT in five-level and six-level atomic systems, which demonstrate steady-state population inversion from selective nonlinear excitation. We will also present studies of resonant hyper-Raman and four-wave mixing processes that are enhanced via suppression of the lower-order linear and nonlinear absorptions, and are capable of generating short-wavelength, coherent light at low pump intensities.

  17. In vivo non-invasive multiphoton tomography of human skin

    NASA Astrophysics Data System (ADS)

    König, Karsten; Riemann, Iris; Ehlers, Alexander; Le Harzic, Ronan

    2005-10-01

    High resolution non-invasive 3D imaging devices are required to detect pathogenic microorganisms such as Anthrax spores, bacteria, viruses, fungi and chemical agents entering biological tissues such as the epidermis. Due to the low light penetration depth and the biodamage potential, ultraviolet light sources can not be employed to realize intratissue imaging of bio- and chemohazards. We report on the novel near infrared laser technology multiphoton tomography and the high resolution 4D imaging tool DermaInspect for non-invasive detection of intratissue agents and their influence on cellular metabolism based on multiphoton autofluorescence imaging (MAI) and second harmonic generation (SHG). Femtosecond laser pulses in the spectral range of 750 nm to 850 nm have been used to image in vivo human skin with subcellular spatial and picosecond temporal resolution. The non-linear induced autofluorescence of both, skin tissues and microorganisms, originates mainly from naturally endogenous fluorophores/protein structures like NAD(P)H, flavins, keratin, collagen, elastin, porphyrins and melanin. Bacteria emit in the blue/green spectral range due to NAD(P)H and flavoproteins and, in certain cases, in the red spectral range due to the biosynthesis of Zn-porphyrins, coproporphyrin and protoporphyrin. Collagen and exogenous non-centrosymmetric molecules can be detected by SHG signals. The system DermaInspect consists of a wavelength-tunable compact 80/90 MHz Ti:sapphire laser, a scan module with galvo scan mirrors, piezo-driven objective, fast photon detector and time-resolved single photon counting unit. It can be used to perform optical sectioning and 3D autofluorescence lifetime imaging (τ-mapping) with 1 μm spatial resolution and 270 ps temporal resolution. The parameter fluorescence lifetime depends on the type of fluorophore and its microenvironment and can be used to distinguish bio- and chemohazards from cellular background and to gain information for pathogen

  18. Spectral-overlap approach to multiframe superresolution image reconstruction.

    PubMed

    Cohen, Edward; Picard, Richard H; Crabtree, Peter N

    2016-05-20

    Various techniques and algorithms have been developed to improve the resolution of sensor-aliased imagery captured with multiple subpixel-displaced frames on an undersampled pixelated image plane. These dealiasing algorithms are typically known as multiframe superresolution (SR), or geometric SR to emphasize the role of the focal-plane array. Multiple low-resolution (LR) aliased frames of the same scene are captured and allocated to a common high-resolution (HR) reconstruction grid, leading to the possibility of an alias-free reconstruction, as long as the HR sampling rate is above the Nyquist rate. Allocating LR-frame irradiances to HR frames requires the use of appropriate weights. Here we present a novel approach in the spectral domain to calculating exactly weights based on spatial overlap areas, which we call the spectral-overlap (SO) method. We emphasize that the SO method is not a spectral approach but rather an approach to calculating spatial weights that uses spectral decompositions to exploit the array properties of the HR and LR pixels. The method is capable of dealing with arbitrary aliasing factors and interframe motions consisting of in-plane translations and rotations. We calculate example reconstructed HR images (the inverse problem) from synthetic aliased images for integer and for fractional aliasing factors. We show the utility of the SO-generated overlap-area weights in both noniterative and iterative reconstructions with known or unknown aliasing factor. We show how the overlap weights can be used to generate the Green's function (pixel response function) for noniterative dealiasing. In addition, we show how the overlap-area weights can be used to generate synthetic aliased images (the forward problem). We compare the SO approach to the spatial-domain geometric approach of O'Rourke and find virtually identical high accuracy but with significant enhancements in speed for SO. We also compare the SO weights to interpolated weights and find that

  19. Multiphoton Microscopy for Visualizing Lipids in Tissue.

    PubMed

    Lee, Martin; Serrels, Alan

    2016-01-01

    Visualizing the appearance of fat droplets and adipocytes in tissue can be realized using a label-free imaging method known as coherent anti-Stokes Raman spectroscopy (CARS). CARS is a nonlinear optical technique that allows label-free imaging of a material with contrast based on the same vibrational signatures of molecules found in Raman spectroscopy. CARS can be combined with other single and multiphoton imaging modes such as second harmonic generation and two-photon fluorescence to image a broad variety of biological structures.Here we describe the construction of a multiphoton microscope that will enable the study of both fluorescently labeled and unlabeled tissue. This has been used to monitor the contribution of Wt1 expressing cells towards the visceral fat depots during gestation. PMID:27417963

  20. Spectral-Spatial Classification of Hyperspectral Images Using Hierarchical Optimization

    NASA Technical Reports Server (NTRS)

    Tarabalka, Yuliya; Tilton, James C.

    2011-01-01

    A new spectral-spatial method for hyperspectral data classification is proposed. For a given hyperspectral image, probabilistic pixelwise classification is first applied. Then, hierarchical step-wise optimization algorithm is performed, by iteratively merging neighboring regions with the smallest Dissimilarity Criterion (DC) and recomputing class labels for new regions. The DC is computed by comparing region mean vectors, class labels and a number of pixels in the two regions under consideration. The algorithm is converged when all the pixels get involved in the region merging procedure. Experimental results are presented on two remote sensing hyperspectral images acquired by the AVIRIS and ROSIS sensors. The proposed approach improves classification accuracies and provides maps with more homogeneous regions, when compared to previously proposed classification techniques.

  1. Imaging of blood cells based on snapshot Hyper-Spectral Imaging systems

    NASA Astrophysics Data System (ADS)

    Robison, Christopher J.; Kolanko, Christopher; Bourlai, Thirimachos; Dawson, Jeremy M.

    2015-05-01

    Snapshot Hyper-Spectral imaging systems are capable of capturing several spectral bands simultaneously, offering coregistered images of a target. With appropriate optics, these systems are potentially able to image blood cells in vivo as they flow through a vessel, eliminating the need for a blood draw and sample staining. Our group has evaluated the capability of a commercial Snapshot Hyper-Spectral imaging system, the Arrow system from Rebellion Photonics, in differentiating between white and red blood cells on unstained blood smear slides. We evaluated the imaging capabilities of this hyperspectral camera; attached to a microscope at varying objective powers and illumination intensity. Hyperspectral data consisting of 25, 443x313 hyperspectral bands with ~3nm spacing were captured over the range of 419 to 494nm. Open-source hyper-spectral data cube analysis tools, used primarily in Geographic Information Systems (GIS) applications, indicate that white blood cells features are most prominent in the 428-442nm band for blood samples viewed under 20x and 50x magnification over a varying range of illumination intensities. These images could potentially be used in subsequent automated white blood cell segmentation and counting algorithms for performing in vivo white blood cell counting.

  2. Feasibility study of a novel miniaturized spectral imaging system architecture in UAV surveillance

    NASA Astrophysics Data System (ADS)

    Liu, Shuyang; Zhou, Tao; Jia, Xiaodong; Cui, Hushan; Huang, Chengjun

    2016-01-01

    The spectral imaging technology is able to analysis the spectral and spatial geometric character of the target at the same time. To break through the limitation brought by the size, weight and cost of the traditional spectral imaging instrument, a miniaturized novel spectral imaging based on CMOS processing has been introduced in the market. This technology has enabled the possibility of applying spectral imaging in the UAV platform. In this paper, the relevant technology and the related possible applications have been presented to implement a quick, flexible and more detailed remote sensing system.

  3. Spectral photoplethysmographic imaging sensor fusion for enhanced heart rate detection

    NASA Astrophysics Data System (ADS)

    Amelard, Robert; Clausi, David A.; Wong, Alexander

    2016-03-01

    Continuous heart rate monitoring can provide important context for quantitative clinical assessment in scenarios such as long-term health monitoring and disability prevention. Photoplethysmographic imaging (PPGI) systems are particularly useful for such monitoring scenarios as contact-based devices pose problems related to comfort and mobility. Each pixel can be regarded as a virtual PPG sensor, thus enabling simultaneous measurements of multiple skin sites. Existing PPGI systems analyze temporal PPGI sensor uctuations related to hemodynamic pulsations across a region of interest to extract the blood pulse signal. However, due to spatially varying optical properties of the skin, the blood pulse signal may not be consistent across all PPGI sensors, leading to inaccurate heart rate monitoring. To increase the hemodynamic signal-to-noise ratio (SNR), we propose a novel spectral PPGI sensor fusion method for enhanced estimation of the true blood pulse signal. Motivated by the observation that PPGI sensors with high hemodynamic SNR exhibit a spectral energy peak at the heart rate frequency, an entropy-based fusion model was formulated to combine PPGI sensors based on the sensors' spectral energy distribution. The optical PPGI device comprised a near infrared (NIR) sensitive camera and an 850 nm LED. Spatially uniform irradiance was achieved by placing optical elements along the LED beam, providing consistent illumination across the skin area. Dual-mode temporally coded illumination was used to negate the temporal effect of ambient illumination. Experimental results show that the spectrally weighted PPGI method can accurately and consistently extract heart rate information where traditional region-based averaging fails.

  4. Open-box spectral clustering: applications to medical image analysis.

    PubMed

    Schultz, Thomas; Kindlmann, Gordon L

    2013-12-01

    Spectral clustering is a powerful and versatile technique, whose broad range of applications includes 3D image analysis. However, its practical use often involves a tedious and time-consuming process of tuning parameters and making application-specific choices. In the absence of training data with labeled clusters, help from a human analyst is required to decide the number of clusters, to determine whether hierarchical clustering is needed, and to define the appropriate distance measures, parameters of the underlying graph, and type of graph Laplacian. We propose to simplify this process via an open-box approach, in which an interactive system visualizes the involved mathematical quantities, suggests parameter values, and provides immediate feedback to support the required decisions. Our framework focuses on applications in 3D image analysis, and links the abstract high-dimensional feature space used in spectral clustering to the three-dimensional data space. This provides a better understanding of the technique, and helps the analyst predict how well specific parameter settings will generalize to similar tasks. In addition, our system supports filtering outliers and labeling the final clusters in such a way that user actions can be recorded and transferred to different data in which the same structures are to be found. Our system supports a wide range of inputs, including triangular meshes, regular grids, and point clouds. We use our system to develop segmentation protocols in chest CT and brain MRI that are then successfully applied to other datasets in an automated manner.

  5. Gemstone spectral imaging for measuring adult bone mineral density

    PubMed Central

    Shao, Wei-Guang; Liu, Dian-Mei

    2016-01-01

    The present study aimed to detect the bone Ca2+ content of L3 vertebrae in adults by gemstone spectral computed tomography. In total, 235 patients were selected and divided into age groups of 10 years each. The scanning data were used to detect the water-based and Ca2+-based substance levels on the L3 vertebral cancellous bone images. The results indicated that there were significant differences in vertebral Ca2+-water and water-Ca2+ densities determined by gemstone spectral imaging (GSI) between males and females in subjects aged 50–59 years, 60–69 years, 70–79 years and ≥80 years (P<0.05). The ages of male and female participants were negatively correlated with vertebral Ca2+-water density (P<0.01) and water-Ca2+ density (P<0.01). In conclusion, GSI may be used as a novel method of measuring the vertebral adult bone mineral density. PMID:27703518

  6. Imaging spectral electrical properties of variably saturated porous media

    NASA Astrophysics Data System (ADS)

    Kelter, Matthias; Huisman, Johann A.; Kemna, Andreas; Zimmermann, Egon; Vereecken, Harry

    2013-04-01

    The spatial distribution of unsaturated hydraulic conductivity in the subsurface is of importance for hydrological modeling. Conventional methods to determine unsaturated hydraulic properties in the field are invasive and typically have a poor spatial resolution. In order to overcome these drawbacks, geophysical methods have received much attention in the last decades. Recent results of electrical impedance spectroscopy (EIS) on a range of saturated and unsaturated porous media revealed promising relationships between spectral electrical and hydraulic properties. Therefore, spectral electrical impedance tomography (EIT) is a promising method to image hydraulic properties in the subsurface. While this approach is emerging for aquifer characterization, unsaturated hydraulic properties have not yet been determined by EIT. In order to do so, a laboratory setup has been developed to perform controlled infiltration, drainage and stationary flow experiments on soil columns. A lysimeter with a height of 50 cm and a diameter of 22 cm is equipped with 40 electrodes and 4 tensiometers. An irrigation device at the top controlled by a peristaltic pump is used for a constant and homogeneous infiltration. Outflow is controlled by a suction plate at the bottom where an adjustable vacuum of up to 500 hPa can be applied. In a first measurement series, spectral EIT measurements were performed on a homogeneous sand column during stepwise drainage of the saturated porous medium using predefined pressure at the bottom. First results show that with decreasing water content the low frequency phase shift of complex electrical conductivity increases. This is consistent with previously reported EIS results. Calibrated relationships between electrical and hydraulic properties were used to convert the resulting electrical into hydraulic conductivity images.

  7. Double-arm three-dimensional ion imaging apparatus for the study of ion pair channels in resonance enhanced multiphoton ionization.

    PubMed

    Poretskiy, M S; Chichinin, A I; Maul, C; Gericke, K-H

    2016-02-01

    We present a novel experimental configuration for the full quantitative characterization of the multichannel resonance enhanced multiphoton ionization (REMPI) of small molecules in cases when the ion-pair dissociation channel is important. For this purpose, a double-arm time-of-flight mass spectrometer with three-dimensional (3D) ion imaging detectors at both arms is constructed. The REMPI of HCl molecules is used to examine the constructed setup. The apparatus allows us to perform simultaneous measurements of the 3D velocity vector distributions of positive (H(+), HCl(+), and Cl(+)) and negative (Cl(-)) photoions. The characterization consists of the determination of "two-photon absorption cross sections" for the process HCl(X)+2hν → HCl*, one-photon absorption cross sections for subsequent processes HCl* + hν → HCl*, and the probability of the subsequent non-adiabatic transition HCl* → HCl(B) → H(+) + Cl(-), which leads to ionic pairs. All these data should be obtained from the analysis of the dependencies of the number of ions on the laser energy. The full characterization of the laser beam and the knowledge of the ion detection probability are necessary parts of the analysis. Detailed knowledge of losses of produced ions in the mass spectrometer before detection requires understanding and characterization of such processes like electron emission from metallic grids under ion bombardment or charge transfer between positive ions and the metal surface of the grids, like Cl(+) + (grid) → Cl(-). These important phenomena from surface science are rarely discussed in the imaging literature, and here, we try to compensate for this shortcoming. PMID:26931834

  8. Ratiometric spectral imaging for fast tumor detection and chemotherapy monitoring in vivo

    NASA Astrophysics Data System (ADS)

    Hwang, Jae Youn; Gross, Zeev; Gray, Harry B.; Medina-Kauwe, Lali K.; Farkas, Daniel L.

    2011-06-01

    We report a novel in vivo spectral imaging approach to cancer detection and chemotherapy assessment. We describe and characterize a ratiometric spectral imaging and analysis method and evaluate its performance for tumor detection and delineation by quantitatively monitoring the specific accumulation of targeted gallium corrole (HerGa) into HER2-positive (HER2 +) breast tumors. HerGa temporal accumulation in nude mice bearing HER2 + breast tumors was monitored comparatively by a. this new ratiometric imaging and analysis method; b. established (reflectance and fluorescence) spectral imaging; c. more commonly used fluorescence intensity imaging. We also tested the feasibility of HerGa imaging in vivo using the ratiometric spectral imaging method for tumor detection and delineation. Our results show that the new method not only provides better quantitative information than typical spectral imaging, but also better specificity than standard fluorescence intensity imaging, thus allowing enhanced in vivo outlining of tumors and dynamic, quantitative monitoring of targeted chemotherapy agent accumulation into them.

  9. Color camera computed tomography imaging spectrometer for improved spatial-spectral image accuracy

    NASA Technical Reports Server (NTRS)

    Wilson, Daniel W. (Inventor); Bearman, Gregory H. (Inventor); Johnson, William R. (Inventor)

    2011-01-01

    Computed tomography imaging spectrometers ("CTIS"s) having color focal plane array detectors are provided. The color FPA detector may comprise a digital color camera including a digital image sensor, such as a Foveon X3.RTM. digital image sensor or a Bayer color filter mosaic. In another embodiment, the CTIS includes a pattern imposed either directly on the object scene being imaged or at the field stop aperture. The use of a color FPA detector and the pattern improves the accuracy of the captured spatial and spectral information.

  10. Miniature Compressive Ultra-spectral Imaging System Utilizing a Single Liquid Crystal Phase Retarder

    NASA Astrophysics Data System (ADS)

    August, Isaac; Oiknine, Yaniv; Abuleil, Marwan; Abdulhalim, Ibrahim; Stern, Adrian

    2016-03-01

    Spectroscopic imaging has been proved to be an effective tool for many applications in a variety of fields, such as biology, medicine, agriculture, remote sensing and industrial process inspection. However, due to the demand for high spectral and spatial resolution it became extremely challenging to design and implement such systems in a miniaturized and cost effective manner. Using a Compressive Sensing (CS) setup based on a single variable Liquid Crystal (LC) retarder and a sensor array, we present an innovative Miniature Ultra-Spectral Imaging (MUSI) system. The LC retarder acts as a compact wide band spectral modulator. Within the framework of CS, a sequence of spectrally modulated images is used to recover ultra-spectral image cubes. Using the presented compressive MUSI system, we demonstrate the reconstruction of gigapixel spatio-spectral image cubes from spectral scanning shots numbering an order of magnitude less than would be required using conventional systems.

  11. Miniature Compressive Ultra-spectral Imaging System Utilizing a Single Liquid Crystal Phase Retarder

    PubMed Central

    August, Isaac; Oiknine, Yaniv; AbuLeil, Marwan; Abdulhalim, Ibrahim; Stern, Adrian

    2016-01-01

    Spectroscopic imaging has been proved to be an effective tool for many applications in a variety of fields, such as biology, medicine, agriculture, remote sensing and industrial process inspection. However, due to the demand for high spectral and spatial resolution it became extremely challenging to design and implement such systems in a miniaturized and cost effective manner. Using a Compressive Sensing (CS) setup based on a single variable Liquid Crystal (LC) retarder and a sensor array, we present an innovative Miniature Ultra-Spectral Imaging (MUSI) system. The LC retarder acts as a compact wide band spectral modulator. Within the framework of CS, a sequence of spectrally modulated images is used to recover ultra-spectral image cubes. Using the presented compressive MUSI system, we demonstrate the reconstruction of gigapixel spatio-spectral image cubes from spectral scanning shots numbering an order of magnitude less than would be required using conventional systems. PMID:27004447

  12. Experimental demonstration of an adaptive architecture for direct spectral imaging classification.

    PubMed

    Dunlop-Gray, Matthew; Poon, Phillip K; Golish, Dathon; Vera, Esteban; Gehm, Michael E

    2016-08-01

    Spectral imaging is a powerful tool for providing in situ material classification across a spatial scene. Typically, spectral imaging analyses are interested in classification, though often the classification is performed only after reconstruction of the spectral datacube. We present a computational spectral imaging system, the Adaptive Feature-Specific Spectral Imaging Classifier (AFSSI-C), which yields direct classification across the spatial scene without reconstruction of the source datacube. With a dual disperser architecture and a programmable spatial light modulator, the AFSSI-C measures specific projections of the spectral datacube which are generated by an adaptive Bayesian classification and feature design framework. We experimentally demonstrate multiple order-of-magnitude improvement of classification accuracy in low signal-to-noise (SNR) environments when compared to legacy spectral imaging systems.

  13. Spectral imager based on Fabry-Perot interferometer for Aalto-1 nanosatellite

    NASA Astrophysics Data System (ADS)

    Mannila, Rami; Näsilä, Antti; Viherkanto, Kai; Holmlund, Christer; Näkki, Ismo; Saari, Heikki

    2013-09-01

    The Aalto-1 is a 3U-cubesat project coordinated by Aalto University. The satellite, Aalto-1, will be mainly built by students as project assignments and thesis works. The Aalto-1 is planned to launch on 2014. VTT Technical Research Centre of Finland is developing the main Earth observation payload, a miniaturized spectral imager unit, for the satellite. The spectral imager unit contains a spectral imager, a visible RGB-camera and control electronics of the cameras. Detailed design of the spectral imager unit has been completed and assembly of the spectral imager unit will be done in the autumn 2013. The spectral imager is based on a tunable Fabry-Perot interferometer (FPI) accompanied by an RGB CMOS image sensor. The FPI consists of two highly reflective surfaces separated by a tunable air gap and it is based on a piezo-actuated structure. The piezo-actuated FPI uses three piezo-actuators and is controlled in a closed capacitive feedback loop. The spectral resolution of the imager will be 8-15 nm at full width at half maximum and it will operate in the wavelength range 500-900 nm. Imaging resolution of the spectral imager is 1024x1024 pixels and the focal length of the optics is 32 mm and F-number is 3.4. Mass of the spectral imager unit is approximately 600 grams, and dimensions are 97 mm x 97 mm x 48 mm.

  14. Imaging spectral signature satellite instrument for the real-time identification of ground scenes with a dedicated spectral signature

    NASA Astrophysics Data System (ADS)

    Kantojärvi, Uula; Saari, Heikki; Viherkanto, Kai; Herrala, Esko; Harnisch, Bernd

    2007-05-01

    With hyperspectral pushbroom imaging spectrometers on Earth observation satellites it is possible to detect and identify dedicated ground pixels by their spectral signature. Conventional time consuming on-ground processing performs this selection by processing the measured hyperspectral data cube of the image. The Imaging Spectral Signature Instrument (ISSI) concept combines an optical on-board processing of the hyperspectral data cube with a thresholding algorithm, to identify pixels with a pre-defined and programmable spectral signature, such as water, forest and minerals, in the ground swath. The Imaging Spectral Signature Instrument consists of an imaging telescope, which images an object line on the entrance slit of a first imaging spectrometer, which disperses each pixel of the object line into its spectral content and images the hyperspectral image on the spatial light modulator. This spatial light modulator will be programmed with a spatial transmission or reflection behavior, which is constant along the spatial pixels and along the spectral pixels identical to a filter vector that corresponds to the spectral signature of the searched specific feature. A second inverted spectrometer reimages the by the first spectrometer dispersed and by the spatial light modulator transmitted or reflected flux into a line of pixels. In case the spectral content of the ground scene is identical to the searched signature, the flux traversing or reflecting the spatial light modulator will be maximum. The related pixel can be identified in the final image as a high signal by a threshold discriminator. A component test setup consists of an imaging lens, two Imspector™ spectrographs, a spatial light modulator, which is a programmable transmissible liquid crystal display and a CCD sensor as a detector. A mathematical model was developed for the instrument and its performance was evaluated in order to compare different concept variations. All components were measured and

  15. An excitation wavelength-scanning spectral imaging system for preclinical imaging

    NASA Astrophysics Data System (ADS)

    Leavesley, Silas; Jiang, Yanan; Patsekin, Valery; Rajwa, Bartek; Robinson, J. Paul

    2008-02-01

    Small-animal fluorescence imaging is a rapidly growing field, driven by applications in cancer detection and pharmaceutical therapies. However, the practical use of this imaging technology is limited by image-quality issues related to autofluorescence background from animal tissues, as well as attenuation of the fluorescence signal due to scatter and absorption. To combat these problems, spectral imaging and analysis techniques are being employed to separate the fluorescence signal from background autofluorescence. To date, these technologies have focused on detecting the fluorescence emission spectrum at a fixed excitation wavelength. We present an alternative to this technique, an imaging spectrometer that detects the fluorescence excitation spectrum at a fixed emission wavelength. The advantages of this approach include increased available information for discrimination of fluorescent dyes, decreased optical radiation dose to the animal, and ability to scan a continuous wavelength range instead of discrete wavelength sampling. This excitation-scanning imager utilizes an acousto-optic tunable filter (AOTF), with supporting optics, to scan the excitation spectrum. Advanced image acquisition and analysis software has also been developed for classification and unmixing of the spectral image sets. Filtering has been implemented in a single-pass configuration with a bandwidth (full width at half maximum) of 16nm at 550nm central diffracted wavelength. We have characterized AOTF filtering over a wide range of incident light angles, much wider than has been previously reported in the literature, and we show how changes in incident light angle can be used to attenuate AOTF side lobes and alter bandwidth. A new parameter, in-band to out-of-band ratio, was defined to assess the quality of the filtered excitation light. Additional parameters were measured to allow objective characterization of the AOTF and the imager as a whole. This is necessary for comparing the

  16. Spectrally Resolved Magnetic Resonance Imaging of the XenonBiosensor

    SciTech Connect

    Hilty, Christian; Lowery, Thomas; Wemmer, David; Pines, Alexander

    2005-07-15

    Due to its ability to non-invasively record images, as well as elucidate molecular structure, nuclear magnetic resonance is the method of choice for applications as widespread as chemical analysis and medical diagnostics. Its detection threshold is, however, limited by the small polarization of nuclear spins in even the highest available magnetic fields. This limitation can, under certain circumstances, be alleviated by using hyper-polarized substances. Xenon biosensors make use of the sensitivity gain of hyperpolarized xenon to provide magnetic resonance detection capability for a specific low-concentration target. They consist of a cryptophane cage, which binds one xenon atom, and which has been connected via a linker to a targeting moiety such as a ligand or antibody. Recent work has shown the possibility of using the xenon biosensor to detect small amounts of a substance in a heterogeneous environment by NMR. Here, we demonstrate that magnetic resonance (MR) provides the capability to obtain spectrally and spatially resolved images of the distribution of immobilized biosensor, opening the possibility for using the xenon biosensor for targeted imaging.

  17. Terahertz detectors for long wavelength multi-spectral imaging.

    SciTech Connect

    Lyo, Sungkwun Kenneth; Wanke, Michael Clement; Reno, John Louis; Shaner, Eric Arthur; Grine, Albert D.

    2007-10-01

    The purpose of this work was to develop a wavelength tunable detector for Terahertz spectroscopy and imaging. Our approach was to utilize plasmons in the channel of a specially designed field-effect transistor called the grating-gate detector. Grating-gate detectors exhibit narrow-linewidth, broad spectral tunability through application of a gate bias, and no angular dependence in their photoresponse. As such, if suitable sensitivity can be attained, they are viable candidates for Terahertz multi-spectral focal plane arrays. When this work began, grating-gate gate detectors, while having many promising characteristics, had a noise-equivalent power (NEP) of only 10{sup -5} W/{radical}Hz. Over the duration of this project, we have obtained a true NEP of 10{sup -8} W/{radical}Hz and a scaled NEP of 10{sup -9}W/{radical}Hz. The ultimate goal for these detectors is to reach a NEP in the 10{sup -9{yields}-10}W/{radical}Hz range; we have not yet seen a roadblock to continued improvement.

  18. Compressive spectral polarization imaging with coded micropolarizer array

    NASA Astrophysics Data System (ADS)

    Fu, Chen; Arguello, Henry; Sadler, Brian M.; Arce, Gonzalo R.

    2015-05-01

    We present a compressive spectral polarization imager based on a prism which is rotated to different angles as the measurement shots are taken, and a colored detector with a micropolarizer array. The prism shears the scene along one spatial axis according to its wavelength components. The scene is then projected to different locations on the detector as measurement shots are taken. Composed of 0°, 45°, 90°, 135° linear micropolarizers, the pixels of the micropolarizer array matched to that of the colored detector, thus the first three Stokes parameters of the scene are compressively sensed. The four dimensional (4D) data cube is thus projected onto the two dimensional (2D) FPA. Designed patterns for the micropolarizer and the colored detector are applied so as to improve the reconstruction problem. The 4D spectral-polarization data cube is reconstructed from the 2D measurements via nonlinear optimization with sparsity constraints. Computer simulations are performed and the performance of designed patterns is compared with random patterns.

  19. Research on method of geometry and spectral calibration of pushbroom dispersive hyperspectral imager

    NASA Astrophysics Data System (ADS)

    He, Zhiping; Shu, Rong; Wang, Jianyu

    2012-11-01

    Development and application of airborne and aerospace hyperspectral imager press for high precision geometry and spectral calibration of pixels of image cube. The research of geometry and spectral calibration of pushbroom hyperspectral imager, its target is giving the coordinate of angle field of view and center wavelength of each detect unit in focal plane detector of hyperspectral imager, and achieves the high precision, full field of view, full channel geometry and spectral calibration. It is importance for imaging quantitative and deep application of hyperspectal imager. The paper takes the geometry and spectral calibration of pushbroom dispersive hyperspectral imager as case study, and research on the constitution and analysis of imaging mathematical model. Aimed especially at grating-dispersive hyperspectral imaging, the specialty of the imaging mode and dispersive method has been concretely analyzed. Based on the analysis, the theory and feasible method of geometry and spectral calibration of dispersive hyperspectral imager is set up. The key technique has been solved is As follows: 1). the imaging mathematical model and feasible method of geometry and spectral calibration for full pixels of image cube has been set up, the feasibility of the calibration method has been analyzed. 2). the engineering model and method of the geometry and spectral calibration of pushbroom dispersive hyperspectral imager has been set up and the calibration equipment has been constructed, and the calibration precision has been analyzed.

  20. Martian spectral units derived from ISM imaging spectrometer data

    NASA Technical Reports Server (NTRS)

    Murchie, S.; Mustard, J.; Saylor, R.

    1993-01-01

    Based on results of the Viking mission, the soil layer of Mars has been thought to be fairly homogeneous and to consist of a mixture of as few as two components, a 'dark gray' basaltic material and a 'bright red' altered material. However, near-infrared reflectance spectra measured recently both telescopically and from spacecraft indicate compositional heterogeneity beyond what can be explained by just two components. In particular, data from the ISM imaging spectrometer, which observed much of the equatorial region at a spatial resolution of approximately 22 km, indicate spatial differences in the presence and abundance of Fe-containing phases, hydroxylated silicates, and H2O. The ISM data was used to define, characterize, and map soil 'units' based on their spectral properties. The spatial distribution of these 'units' were compared to morphologic, visible color, and thermal inertia features recognized in Viking data.

  1. Unsupervised Learning of Cone Spectral Classes from Natural Images

    PubMed Central

    Benson, Noah C.; Manning, Jeremy R.; Brainard, David H.

    2014-01-01

    The first step in the evolution of primate trichromatic color vision was the expression of a third cone class not present in ancestral mammals. This observation motivates a fundamental question about the evolution of any sensory system: how is it possible to detect and exploit the presence of a novel sensory class? We explore this question in the context of primate color vision. We present an unsupervised learning algorithm capable of both detecting the number of spectral cone classes in a retinal mosaic and learning the class of each cone using the inter-cone correlations obtained in response to natural image input. The algorithm's ability to classify cones is in broad agreement with experimental evidence about functional color vision for a wide range of mosaic parameters, including those characterizing dichromacy, typical trichromacy, anomalous trichromacy, and possible tetrachromacy. PMID:24967877

  2. Unsupervised learning of cone spectral classes from natural images.

    PubMed

    Benson, Noah C; Manning, Jeremy R; Brainard, David H

    2014-06-01

    The first step in the evolution of primate trichromatic color vision was the expression of a third cone class not present in ancestral mammals. This observation motivates a fundamental question about the evolution of any sensory system: how is it possible to detect and exploit the presence of a novel sensory class? We explore this question in the context of primate color vision. We present an unsupervised learning algorithm capable of both detecting the number of spectral cone classes in a retinal mosaic and learning the class of each cone using the inter-cone correlations obtained in response to natural image input. The algorithm's ability to classify cones is in broad agreement with experimental evidence about functional color vision for a wide range of mosaic parameters, including those characterizing dichromacy, typical trichromacy, anomalous trichromacy, and possible tetrachromacy. PMID:24967877

  3. Unsupervised learning of cone spectral classes from natural images.

    PubMed

    Benson, Noah C; Manning, Jeremy R; Brainard, David H

    2014-06-01

    The first step in the evolution of primate trichromatic color vision was the expression of a third cone class not present in ancestral mammals. This observation motivates a fundamental question about the evolution of any sensory system: how is it possible to detect and exploit the presence of a novel sensory class? We explore this question in the context of primate color vision. We present an unsupervised learning algorithm capable of both detecting the number of spectral cone classes in a retinal mosaic and learning the class of each cone using the inter-cone correlations obtained in response to natural image input. The algorithm's ability to classify cones is in broad agreement with experimental evidence about functional color vision for a wide range of mosaic parameters, including those characterizing dichromacy, typical trichromacy, anomalous trichromacy, and possible tetrachromacy.

  4. Fully Quantified Spectral Imaging Reveals in Vivo Membrane Protein Interactions

    PubMed Central

    King, Christopher; Stoneman, Michael; Raicu, Valerica; Hristova, Kalina

    2016-01-01

    Here we introduce the Fully Quantified Spectral Imaging (FSI) method as a new tool to probe the stoichiometry and stability of protein complexes in biological membranes. The FSI method yields two dimensional membrane concentrations and FRET efficiencies in native plasma membranes. It can be used to characterize the association of membrane proteins: to differentiate between monomers, dimers, or oligomers, to produce binding (association) curves, and to measure the free energies of association in the membrane. We use the FSI method to study the lateral interactions of Vascular Endothelial Growth Factor Receptor 2 (VEGFR2), a member of the receptor tyrosine kinase (RTK) superfamily, in plasma membranes, in vivo. The knowledge gained through the use of the new method challenges the current understanding of VEGFR2 signaling. PMID:26787445

  5. Superlocalization spectral imaging microscopy of a multicolor quantum dot complex.

    PubMed

    Shi, Xingbo; Xie, Zhongqiu; Song, Yuehong; Tan, Yongjun; Yeung, Edward S; Gai, Hongwei

    2012-02-01

    The key factor of realizing super-resolution optical microscopy at the single-molecule level is to separately position two adjacent molecules. An opportunity to independently localize target molecules is provided by the intermittency (blinking) in fluorescence of a quantum dot (QD) under the condition that the blinking of each emitter can be recorded and identified. Herein we develop a spectral imaging based color nanoscopy which is capable of determining which QD is blinking in the multicolor QD complex through tracking the first-order spectrum, and thus, the distance at tens of nanometers between two QDs is measured. Three complementary oligonucleotides with lengths of 15, 30, and 45 bp are constructed as calibration rulers. QD585 and QD655 are each linked at one end. The measured average distances are in good agreement with the calculated lengths with a precision of 6 nm, and the intracellular dual-color QDs within a diffraction-limited spot are distinguished.

  6. Method for detection and imaging over a broad spectral range

    DOEpatents

    Yefremenko, Volodymyr; Gordiyenko, Eduard; Pishko, legal representative, Olga; Novosad, Valentyn; Pishko, deceased; Vitalii

    2007-09-25

    A method of controlling the coordinate sensitivity in a superconducting microbolometer employs localized light, heating or magnetic field effects to form normal or mixed state regions on a superconducting film and to control the spatial location. Electron beam lithography and wet chemical etching were applied as pattern transfer processes in epitaxial Y--Ba--Cu--O films. Two different sensor designs were tested: (i) a 3 millimeter long and 40 micrometer wide stripe and (ii) a 1.25 millimeters long, and 50 micron wide meandering-like structure. Scanning the laser beam along the stripe leads to physical displacement of the sensitive area, and, therefore, may be used as a basis for imaging over a broad spectral range. Forming the superconducting film as a meandering structure provides the equivalent of a two-dimensional detector array. Advantages of this approach are simplicity of detector fabrication, and simplicity of the read-out process requiring only two electrical terminals.

  7. Interferometric and nonlinear-optical spectral-imaging techniques for outer space and live cells

    NASA Astrophysics Data System (ADS)

    Itoh, Kazuyoshi

    2015-12-01

    Multidimensional signals such as the spectral images allow us to have deeper insights into the natures of objects. In this paper the spectral imaging techniques that are based on optical interferometry and nonlinear optics are presented. The interferometric imaging technique is based on the unified theory of Van Cittert-Zernike and Wiener-Khintchine theorems and allows us to retrieve a spectral image of an object in the far zone from the 3D spatial coherence function. The retrieval principle is explained using a very simple object. The promising applications to space interferometers for astronomy that are currently in progress will also be briefly touched on. An interesting extension of interferometric spectral imaging is a 3D and spectral imaging technique that records 4D information of objects where the 3D and spectral information is retrieved from the cross-spectral density function of optical field. The 3D imaging is realized via the numerical inverse propagation of the cross-spectral density. A few techniques suggested recently are introduced. The nonlinear optical technique that utilizes stimulated Raman scattering (SRS) for spectral imaging of biomedical targets is presented lastly. The strong signals of SRS permit us to get vibrational information of molecules in the live cell or tissue in real time. The vibrational information of unstained or unlabeled molecules is crucial especially for medical applications. The 3D information due to the optical nonlinearity is also the attractive feature of SRS spectral microscopy.

  8. First Radio Burst Imaging Observation From Mingantu Ultrawide Spectral Radioheliograph

    NASA Astrophysics Data System (ADS)

    Yan, Yihua; Chen, Linjie; Yu, Sijie; CSRH Team

    2015-08-01

    Radio imaging spectroscopy over wide range wavelength in dm/cm-bands will open new windows on solar flares and coronal mass ejections by tracing the radio emissions from accelerated electrons. The Chinese Spectral Radioheliograph (CSRH) with two arrays in 400MHz-2GHz /2-15GHz ranges with 64/532 frequency channels have been established in Mingantu Observing Station, Inner Mongolia of China, since 2013 and is in test observations now. CSRH is renamed as MUSER (Mingantu Ultrawide SpEctral Radioheliograph) after it's accomplishment We will introduce the progress and current status of CSRH. Some preliminary results of CSRH will be presented.On 11 Nov2014, the first burst event was registered by MUSER-I array at 400MHz-2GHz waveband. According to SGD event list there was a C-class flare peaked at 04:49UT in the disk center and the radio bursts around 04:22-04:24UT was attributed to this flare. However, MUSER-I image observation of the burst indicates that the radio burst peaked around 04:22UT was due to the eruption at the east limb of the Sun and connected to a CME appeared in that direction about 1 hour later. This demonstrate the importance of the spectroscopy observation of the solar radio burst.Acknowledgement: The CSRH team includes Wei Wang, Zhijun Chen, Fei Liu, Lihong Geng and Jian Zhang and CSRH project is supported by National Major Scientific Equipment R&D Project ZDYZ2009-3. The research was also supported by NSFC grants (11433006, 11221063), MOST grant (MOST2011CB811401), CAS Pilot-B Project (XDB09000000) and Marie Curie PIRSES- GA-295272-RADIOSUN.

  9. Development of a real-time flexible multiphoton microendoscope for label-free imaging in a live animal

    PubMed Central

    Ducourthial, Guillaume; Leclerc, Pierre; Mansuryan, Tigran; Fabert, Marc; Brevier, Julien; Habert, Rémi; Braud, Flavie; Batrin, Renaud; Vever-Bizet, Christine; Bourg-Heckly, Geneviève; Thiberville, Luc; Druilhe, Anne; Kudlinski, Alexandre; Louradour, Frédéric

    2015-01-01

    We present a two-photon microendoscope capable of in vivo label-free deep-tissue high-resolution fast imaging through a very long optical fiber. First, an advanced light-pulse spectro-temporal shaping device optimally precompensates for linear and nonlinear distortions occurring during propagation within the endoscopic fiber. This enables the delivery of sub-40-fs duration infrared excitation pulses at the output of 5 meters of fiber. Second, the endoscopic fiber is a custom-made double-clad polarization-maintaining photonic crystal fiber specifically designed to optimize the imaging resolution and the intrinsic luminescence backward collection. Third, a miniaturized fiber-scanner of 2.2 mm outer diameter allows simultaneous second harmonic generation (SHG) and two-photon excited autofluorescence (TPEF) imaging at 8 frames per second. This microendoscope’s transverse and axial resolutions amount respectively to 0.8 μm and 12 μm, with a field-of-view as large as 450 μm. This microendoscope’s unprecedented capabilities are validated during label-free imaging, ex vivo on various fixed human tissue samples, and in vivo on an anesthetized mouse kidney demonstrating an imaging penetration depth greater than 300 μm below the surface of the organ. The results reported in this manuscript confirm that nonlinear microendoscopy can become a valuable clinical tool for real-time in situ assessment of pathological states. PMID:26673905

  10. Development of a real-time flexible multiphoton microendoscope for label-free imaging in a live animal

    NASA Astrophysics Data System (ADS)

    Ducourthial, Guillaume; Leclerc, Pierre; Mansuryan, Tigran; Fabert, Marc; Brevier, Julien; Habert, Rémi; Braud, Flavie; Batrin, Renaud; Vever-Bizet, Christine; Bourg-Heckly, Geneviève; Thiberville, Luc; Druilhe, Anne; Kudlinski, Alexandre; Louradour, Frédéric

    2015-12-01

    We present a two-photon microendoscope capable of in vivo label-free deep-tissue high-resolution fast imaging through a very long optical fiber. First, an advanced light-pulse spectro-temporal shaping device optimally precompensates for linear and nonlinear distortions occurring during propagation within the endoscopic fiber. This enables the delivery of sub-40-fs duration infrared excitation pulses at the output of 5 meters of fiber. Second, the endoscopic fiber is a custom-made double-clad polarization-maintaining photonic crystal fiber specifically designed to optimize the imaging resolution and the intrinsic luminescence backward collection. Third, a miniaturized fiber-scanner of 2.2 mm outer diameter allows simultaneous second harmonic generation (SHG) and two-photon excited autofluorescence (TPEF) imaging at 8 frames per second. This microendoscope’s transverse and axial resolutions amount respectively to 0.8 μm and 12 μm, with a field-of-view as large as 450 μm. This microendoscope’s unprecedented capabilities are validated during label-free imaging, ex vivo on various fixed human tissue samples, and in vivo on an anesthetized mouse kidney demonstrating an imaging penetration depth greater than 300 μm below the surface of the organ. The results reported in this manuscript confirm that nonlinear microendoscopy can become a valuable clinical tool for real-time in situ assessment of pathological states.

  11. Hybrid reflecting objectives for functional multiphoton microscopy in turbid media

    PubMed Central

    Vučinić, Dejan; Bartol, Thomas M.; Sejnowski, Terrence J.

    2010-01-01

    Most multiphoton imaging of biological specimens is performed using microscope objectives optimized for high image quality under wide-field illumination. We present a class of objectives designed de novo without regard for these traditional constraints, driven exclusively by the needs of fast multiphoton imaging in turbid media: the delivery of femtosecond pulses without dispersion and the efficient collection of fluorescence. We model the performance of one such design optimized for a typical brain-imaging setup and show that it can greatly outperform objectives commonly used for this task. PMID:16880851

  12. [Full-field and automatic methodology of spectral calibration for PGP imaging spectrometer].

    PubMed

    Sun, Ci; Bayanheshig; Cui, Ji-cheng; Pan, Ming-zhong; Li, Xiao-tian; Tang, Yu-guo

    2014-08-01

    In order to analyze spectral data quantitatively which is obtained by prism-grating-prism imaging spectrometer, spectral calibration is required in order to determine spectral characteristics of PGP imaging spectrometer, such as the center wavelength of every spectral channel, spectral resolution and spectral bending. A spectral calibration system of full field based on collimated monochromatic light method is designed. Spherical mirror is used to provide collimated light, and a freely sliding and rotating folding mirror is adopted to change the angle of incident light in order to realize full field and automatic calibration of imaging spectrometer. Experiments of spectral calibration have been done for PGP imaging spectrometer to obtain parameters of spectral performance, and accuracy analysis combined with the structural features of the entire spectral calibration system have been done. Analysis results indicate that spectral calibration accuracy of the calibration system reaches 0.1 nm, and the bandwidth accuracy reaches 1.3%. The calibration system has merits of small size, better commonality, high precision and so on, and because of adopting the control of automation, the additional errors which are caused by human are avoided. The calibration system can be used for spectral calibration of other imaging spectrometers whose structures are similar to PGP.

  13. Methods of foreign fiber detecting based on PCA analyzing of infrared spectral images

    NASA Astrophysics Data System (ADS)

    Tian, LiXun; Fu, WeiSen; Liu, JieYu; Zhang, HongBo; Pan, Jin; Wang, YaPeng; Tong, Fei

    2014-02-01

    A set of near infrared high resolution spectral imaging system is set up, the infrared absorption properties of raw cotton and colorless foreign are analyzed through the system, and scheme of polypropylene fiber detection based on the near infrared spectral image is proposed; On this basis, reduce dimensions the spectral images through the principal component analysis, further improve the efficiency of colorless foreign detection. The experimental results show that the spectral images after reducing dimensions can be used to detect colorless or light color raw cotton fiber effectively.

  14. Multiphoton dissociative ionization of CS+

    NASA Astrophysics Data System (ADS)

    Rajput, Jyoti; Jochim, Bethany; Zohrabi, M.; Betsch, K. J.; Ablikim, U.; Berry, Ben; Severt, T.; Summers, A. M.; Armstrong, G. S. J.; Esry, B. D.; Carnes, K. D.; Ben-Itzhak, I.

    2015-05-01

    We have studied the dissociative photoionization of a CS+ molecular ion beam in the strong-field regime using <50 fs IR laser pulses (λ ~ 790 nm) from a 10 kHz, ~2 mJ (per pulse) Ti:Sapphire laser system. A coincidence three-dimensional momentum imaging method was used to measure all ions and neutrals formed during this multiphoton process. Two prominent channels were observed: charge-symmetric dissociation, yielding C+ + S+, and charge-asymmetric dissociation, yielding C + S2+. The differences between these two channels with reference to their relative production probability, energetics, and angular distributions is the focus of this work. This work was supported by the Chemical Sciences, Geosciences, and Biosciences Division, Office of Basic Energy Sciences, Office of Science, U.S. Department of Energy. BJ is also supported by DOE-SCGF (DE-AC05-06OR23100).

  15. Review of spectral imaging technology in biomedical engineering: achievements and challenges.

    PubMed

    Li, Qingli; He, Xiaofu; Wang, Yiting; Liu, Hongying; Xu, Dongrong; Guo, Fangmin

    2013-10-01

    Spectral imaging is a technology that integrates conventional imaging and spectroscopy to get both spatial and spectral information from an object. Although this technology was originally developed for remote sensing, it has been extended to the biomedical engineering field as a powerful analytical tool for biological and biomedical research. This review introduces the basics of spectral imaging, imaging methods, current equipment, and recent advances in biomedical applications. The performance and analytical capabilities of spectral imaging systems for biological and biomedical imaging are discussed. In particular, the current achievements and limitations of this technology in biomedical engineering are presented. The benefits and development trends of biomedical spectral imaging are highlighted to provide the reader with an insight into the current technological advances and its potential for biomedical research.

  16. First multiphoton tomography of brain in man

    NASA Astrophysics Data System (ADS)

    König, Karsten; Kantelhardt, Sven R.; Kalasauskas, Darius; Kim, Ella; Giese, Alf

    2016-03-01

    We report on the first two-photon in vivo brain tissue imaging study in man. High resolution in vivo histology by multiphoton tomography (MPT) including two-photon FLIM was performed in the operation theatre during neurosurgery to evaluate the feasibility to detect label-free tumor borders with subcellular resolution. This feasibility study demonstrates, that MPT has the potential to identify tumor borders on a cellular level in nearly real-time.

  17. [Spectral curve shape feature-based hyperspectral remote sensing image retrieval].

    PubMed

    Li, Fei; Zhou, Cheng-Hu; Chen, Rong-Guo

    2008-11-01

    With the rapid development of technology of sensors and data transmission, using all kinds of airplane sensors and satellite sensors, the authors can get different voluminous remote sensing image data of earth. Those voluminous remote sensing image data bring problems of data storage and management. It is becoming increasingly necessary to retrieve some information the authors need from those voluminous image data. Image retrieval was proposed by CHANG firstly in 1980 and can be regarded as expansion of traditional information retrieval. Oriented to the demands of efficient retrieval for voluminous remote sensing image, and considering that there are many bands in hyperspectral remote sensing image, the authors first analyzed image distance function and similarity measure in image retrieval. The most crucial issues in retrieval are spectral features extraction and similarity measure. In the present paper, the authors used classical Douglas-Peucker algorithm (hereinafter referred to DP algorithm) for curve simplification to extract shape features of spectral curve, in order to speed up hyperspectral remote sensing image retrieval. And the authors proposed a new method of spectral curve and remote sensing image retrieval, called Douglas-Peucker Spectral Retrieval algorithm (hereinafter referred to DPSR algorithm). Spectral shape features were used in image retrieval. DPSR used features of spectral curve, reduced the computation amount, realized match and retrieval efficiently, and is suitable for spectral curve retrieval in hyperspectral remote sensing image. The authors selected four ground features (grass, apple garden, grape garden and pond) in OMISI hyperspectral remote sensing image to compute similarity measure results, in order to test the effect of DPSR algorithm. Compared with traditional analysis method such as spectral angle match (SAM) and spectral information divergence (SID), DPSR can maintain high precision of results with less amount of computation

  18. Comparison of label-free and GFP multiphoton imaging of hair follicle-associated pluripotent (HAP) stem cells in mouse whiskers.

    PubMed

    Uchugonova, Aisada; Cao, Wenluo; Hoffman, Robert M; Koenig, Karsten

    2015-01-01

    Hair-follicle-associated pluripotent (HAP) stem cells can differentiate into many cell types, including neurons and heart muscle cells, and have been shown to repair peripheral nerves and the spinal cord in mice. HAP stem cells can be obtained from each individual patient for regenerative medicine which overcomes problems with immune rejection. Previously, we have demonstrated that genetically-encoded protein markers such as GFP in transgenic mice can be used to visualize HAP stem cells in vivo by multiphoton tomography. Detection and visualization of stem cells in vivo without exogenous labels such as GFP would be important for human application. In the present report, we demonstrate label-free visualization of hair follicle stem cells in mouse whiskers by multiphoton tomography due to the intrinsic fluorophores such as NAD(P)H/flavins. We compared multiphoton tomography of GFP-labeled HAP stem cells and unlabeled stem cells in isolated mouse whiskers. We show that observation of HAP stem cells by label-free multiphoton tomography is comparable to detection using GFP-labeled stem cells. The results described here have important implications for detection and isolation of human HAP stem cells for regenerative medicine.

  19. Spectral and multispectral imaging studies of lunar mantled mare deposits

    NASA Technical Reports Server (NTRS)

    Blewett, D. T.; Hawke, B. R.; Lucey, P. G.; Bell, J. F., III; Jaumann, R.; Hiesinger, H.; Neukum, G.; Spudis, P. D.

    1993-01-01

    Near-IR reflectance spectra (0.6-2.5 microns) and CCD images in the extended visible range (0.4-1.0 microns) obtained with Earth-based telescopes have been used to investigate the composition and origin of formations in the Schiller-Schickard region of the Moon. Of particular interest are the Schickard light plains, which represent an area of mantled mare basalt, or cryptomare. Here local pre-existing mare basalts were eroded and incorporated into a highlands-rich deposit by eject a from the Orientale Basin. Spectra observations of mature and immature highland and mare surfaces, as well as dark-halo crater materials provide information on the mafic mineralogy of features in the area. Analyses of the '1 micron' absorption band and spectral mixing models indicate that selected spots in the light plains contain on the order of 50 percent mare basalt. CCD image cubes can be used to map the amount of basalt in the light plains and evaluate changes with radial distance from Orientale.

  20. Retinal Imaging of Infants on Spectral Domain Optical Coherence Tomography

    PubMed Central

    Vinekar, Anand; Mangalesh, Shwetha; Jayadev, Chaitra; Maldonado, Ramiro S.; Bauer, Noel; Toth, Cynthia A.

    2015-01-01

    Spectral domain coherence tomography (SD OCT) has become an important tool in the management of pediatric retinal diseases. It is a noncontact imaging device that provides detailed assessment of the microanatomy and pathology of the infant retina with a short acquisition time allowing office examination without the requirement of anesthesia. Our understanding of the development and maturation of the infant fovea has been enhanced by SD OCT allowing an in vivo assessment that correlates with histopathology. This has helped us understand the critical correlation of foveal development with visual potential in the first year of life and beyond. In this review, we summarize the recent literature on the clinical applications of SD OCT in studying the pathoanatomy of the infant macula, its ability to detect subclinical features, and its correlation with disease and vision. Retinopathy of prematurity and macular edema have been discussed in detail. The review also summarizes the current status of SD OCT in other infant retinal conditions, imaging the optic nerve, the choroid, and the retinal nerve fibre in infants and children, and suggests future areas of research. PMID:26221606

  1. Design and fabrication of the Infrared Spectral Imaging Radiometer (ISIR)

    NASA Astrophysics Data System (ADS)

    Hoffman, James W.; Grush, Ronald C.

    1997-09-01

    The design and fabrication of the infrared spectral imaging radiometer (ISIR) is presented. The ISIR was designed in 1994 to provide calibrated images in four thermal wavelength bands without cryogenic cooling by utilizing the new, uncooled microbolometer detector technology. The complete system was fabricated at Space Instruments, Inc. (SI) in 1995 and 1996 and delivered to NASA Goddard Space Flight Center (GSFC) for flight on the space shuttle in 1997. Photographs of the flight hardware are shown. The ISIR operates in a pushbroom fashion and utilizes real time, digital time delay and integration (TDI) to improve the signal to noise ratio. From a nominal shuttle altitude of 140 nmi, the nadir pixel subtends 240 by 240 meters on the ground. The size of the radiometer is minimized by the elimination of mechanical scan mechanisms and a space radiator. The ISIR instrument utilizes a through-the- optics calibration system to periodically obtain a two-point calibration for each pixel in the detector array. A blackbody with both heating and cooling capability is used to obtain accurate calibration data for both terrestrial and cloudtop measurements. The timeline logic, TDI integration, mechanism control, calibration, and data formatting are performed in the onboard digital processor which utilizes two microprocessors and seven programmable logic devices. The output data is recorded on two, 8 mm tape recorders.

  2. Augmenting Forest Stand Parameters using Landsat TM Spectral Images

    NASA Astrophysics Data System (ADS)

    Reuveni, Y.; Dahan, E.; Anker, Y.; Sprintsin, M.

    2015-12-01

    Forest stand parameters, such as diameter at breast height (DBH), tree height (H), or volume per hectare (V), are imperative for forest resources assessment. Traditional inventory of forest stand parameters, usually based on fieldwork, is often difficult, time-consuming, and expensive, to conduct in large areas. Therefore, estimating forest stand parameters in large areas using traditional inventory approach augmented by satellites data has a significant implication for sustainable forest management and natural resources efficiency. However, obtaining suitable satellite image data for such purpose is a challenging task mainly because of insignificant knowledge between the forest stand parameters and satellite spectral response relationships. Here, we present the use of Landsat Thematic Mapper (TM) spectral responses data for augmenting forest stand parameter obtained from fieldwork at the Lahav Forest, in the Israeli Northern Negev. A new algorithm was developed in order to use all eight TM band when calculating the linear combination which correlates the most to each one of the forest stand parameters. Each linear combination is obtained first for local area inside the entire studied grid and is then fitted using a simple linear polynomial curve to the known forest stand parameter. Once the relationship between the two is characterized by a linear polynomial equation, the TM linear combination local area data is translated to the same equivalent area of the chosen forest stand parameter. At last, we interpolate the entire TM grid using a higher order polynomial fit applied to all the augmented local area combined together to attain full coverage of the desired forest stand parameter.

  3. Temporal and spectral imaging with micro-CT

    SciTech Connect

    Johnston, Samuel M.; Johnson, G. Allan; Badea, Cristian T.

    2012-08-15

    can be used to segment regions containing iodinated blood and compute measures of cardiac function. Conclusions: We believe this combined spectral and temporal imaging technique will be useful for future studies of cardiopulmonary disease in small animals.

  4. Software defined multi-spectral imaging for Arctic sensor networks

    NASA Astrophysics Data System (ADS)

    Siewert, Sam; Angoth, Vivek; Krishnamurthy, Ramnarayan; Mani, Karthikeyan; Mock, Kenrick; Singh, Surjith B.; Srivistava, Saurav; Wagner, Chris; Claus, Ryan; Vis, Matthew Demi

    2016-05-01

    Availability of off-the-shelf infrared sensors combined with high definition visible cameras has made possible the construction of a Software Defined Multi-Spectral Imager (SDMSI) combining long-wave, near-infrared and visible imaging. The SDMSI requires a real-time embedded processor to fuse images and to create real-time depth maps for opportunistic uplink in sensor networks. Researchers at Embry Riddle Aeronautical University working with University of Alaska Anchorage at the Arctic Domain Awareness Center and the University of Colorado Boulder have built several versions of a low-cost drop-in-place SDMSI to test alternatives for power efficient image fusion. The SDMSI is intended for use in field applications including marine security, search and rescue operations and environmental surveys in the Arctic region. Based on Arctic marine sensor network mission goals, the team has designed the SDMSI to include features to rank images based on saliency and to provide on camera fusion and depth mapping. A major challenge has been the design of the camera computing system to operate within a 10 to 20 Watt power budget. This paper presents a power analysis of three options: 1) multi-core, 2) field programmable gate array with multi-core, and 3) graphics processing units with multi-core. For each test, power consumed for common fusion workloads has been measured at a range of frame rates and resolutions. Detailed analyses from our power efficiency comparison for workloads specific to stereo depth mapping and sensor fusion are summarized. Preliminary mission feasibility results from testing with off-the-shelf long-wave infrared and visible cameras in Alaska and Arizona are also summarized to demonstrate the value of the SDMSI for applications such as ice tracking, ocean color, soil moisture, animal and marine vessel detection and tracking. The goal is to select the most power efficient solution for the SDMSI for use on UAVs (Unoccupied Aerial Vehicles) and other drop

  5. Temporal and spectral imaging with micro-CT

    PubMed Central

    Johnston, Samuel M.; Johnson, G. Allan; Badea, Cristian T.

    2012-01-01

    can be used to segment regions containing iodinated blood and compute measures of cardiac function. Conclusions: We believe this combined spectral and temporal imaging technique will be useful for future studies of cardiopulmonary disease in small animals. PMID:22894420

  6. Studies of atmospheric molecules by multiphoton spectroscopy

    SciTech Connect

    Johnson, P.M.

    1991-10-01

    Carbon dioxide presents a great challenge to spectroscopy because of its propensity toward dissociation in all of its excited states. Multiphoton ionization spectroscopy is usually not applicable to the study of dissociating molecules because the dissociation competes effectively with ionization, resulting in no signal. We reasoned, however, that with high enough laser fluence, ionization could compete with dissociation in the longer lived states, exposing them for study from the continuous spectral background resulting from rapidly dissociating states. We describe the various spectroscopic and photophysical effects found through the multiphoton ionization and multiphoton photoelectron spectra. A recently developed variant of threshold ionization spectroscopy, usually called ZEKE, has shown a great deal of usefulness in providing the same information as traditional photoelectron spectroscopy but with higher resolution and much better signal-to-noise when using standard laboratory lasers. Threshold ionization techniques locate the states of an ion by scanning a light source across the ionization continuum of a neutral and somehow detecting when electrons are produced with no kinetic energy. We chose to develop our capabilities in threshold ionization spectroscopy using aromatic molecules because of their importance and because their electronic structure allows a pump-probe type of excitation scheme which avoids the use of vacuum ultraviolet laser beams. Among aromatics, the azines are noted for their small S{sub 1}-T{sub 1} energy gap which give them unique and interesting photophysical properties. We have continued our work on the multiphoton spectrum of metastable nitrogen produced by an electric discharge in supersonic beam. We have been able to assign more of the lines and simulated their rotational structure but many peaks remain unassigned.

  7. Multiphoton light-sheet microscopy using wavelength mixing: fast multicolor imaging of the beating Zebrafish heart with low photobleaching

    NASA Astrophysics Data System (ADS)

    Mahou, Pierre; Vermot, Julien; Beaurepaire, Emmanuel; Supatto, Willy

    2015-03-01

    Two-photon laser scanning microscopy has become a standard to map thick and live tissues. However, its application for fast and multicolor imaging remains challenging. To address this issue, we report on the implementation of mixed wavelength excitation in a two-photon light-sheet microscope. We illustrate the potential of the technique by recording sustained multicolor two-photon movies of the beating heart in zebrafish embryos with negligible photobleaching at 28 million pixels/second. In particular, 3D reconstructions of the heart periodic motion are obtained with sufficient spatiotemporal resolution to track the fast movements of individual cells during a cardiac cycle.

  8. Time-resolved multiphoton imaging of the interaction between the PKC and the NFκB signalling pathways

    NASA Astrophysics Data System (ADS)

    Morton, Penny E.; Ng, Tony; Roberts, Sarah A.; Vojnovic, Borivoj; Ameer-Beg, Simon M.

    2003-10-01

    Using fluorescence resonance energy transfer (FRET) measured by fluorescence lifetime imaging microscopy (FLIM) we have explored the protein-protein interactions between fluorescent protein tagged fusion proteins of the activation pathways of PKC and NFkB. We observe FRET between CFP-IκB and YFP-p65 in unstimulated cells and when treated with TNFα. We also observed a reduction of the fluorescent lifetime of CFP-IκB in the absence of YFP-p65 when TNFα is present.

  9. Fast Multispectral Imaging by Spatial Pixel-Binning and Spectral Unmixing.

    PubMed

    Pan, Zhi-Wei; Shen, Hui-Liang; Li, Chunguang; Chen, Shu-Jie; Xin, John H

    2016-08-01

    Multispectral imaging system is of wide application in relevant fields for its capability in acquiring spectral information of scenes. Its limitation is that, due to the large number of spectral channels, the imaging process can be quite time-consuming when capturing high-resolution (HR) multispectral images. To resolve this limitation, this paper proposes a fast multispectral imaging framework based on the image sensor pixel-binning and spectral unmixing techniques. The framework comprises a fast imaging stage and a computational reconstruction stage. In the imaging stage, only a few spectral images are acquired in HR, while most spectral images are acquired in low resolution (LR). The LR images are captured by applying pixel binning on the image sensor, such that the exposure time can be greatly reduced. In the reconstruction stage, an optimal number of basis spectra are computed and the signal-dependent noise statistics are estimated. Then the unknown HR images are efficiently reconstructed by solving a closed-form cost function that models the spatial and spectral degradations. The effectiveness of the proposed framework is evaluated using real-scene multispectral images. Experimental results validate that, in general, the method outperforms the state of the arts in terms of reconstruction accuracy, with additional 20× or more improvement in computational efficiency.

  10. The Spectral Image Processing System (SIPS) - Interactive visualization and analysis of imaging spectrometer data

    NASA Technical Reports Server (NTRS)

    Kruse, F. A.; Lefkoff, A. B.; Boardman, J. W.; Heidebrecht, K. B.; Shapiro, A. T.; Barloon, P. J.; Goetz, A. F. H.

    1993-01-01

    The Center for the Study of Earth from Space (CSES) at the University of Colorado, Boulder, has developed a prototype interactive software system called the Spectral Image Processing System (SIPS) using IDL (the Interactive Data Language) on UNIX-based workstations. SIPS is designed to take advantage of the combination of high spectral resolution and spatial data presentation unique to imaging spectrometers. It streamlines analysis of these data by allowing scientists to rapidly interact with entire datasets. SIPS provides visualization tools for rapid exploratory analysis and numerical tools for quantitative modeling. The user interface is X-Windows-based, user friendly, and provides 'point and click' operation. SIPS is being used for multidisciplinary research concentrating on use of physically based analysis methods to enhance scientific results from imaging spectrometer data. The objective of this continuing effort is to develop operational techniques for quantitative analysis of imaging spectrometer data and to make them available to the scientific community prior to the launch of imaging spectrometer satellite systems such as the Earth Observing System (EOS) High Resolution Imaging Spectrometer (HIRIS).

  11. Spectrally programmable light engine for in vitro or in vivo molecular imaging and spectroscopy

    NASA Astrophysics Data System (ADS)

    MacKinnon, Nicholas; Stange, Ulrich; Lane, Pierre; Macaulay, Calum; Quatrevalet, Mathieu

    2005-04-01

    A spectrally and temporally programmable light engine can create any spectral profile for hyperspectral, fluorescence, or principal-component imaging or with medical photonics devices employing spectroscopy, microscopy, and endoscopy. Multispectral imaging feasibility was demonstrated by capturing nine images at wavelengths from 450 to 650 mm (25-nm FWHM) with a CCD-camera-equipped bronchoscope coupled to the light engine. Selected wavelength regions were combined to produce a color endoscopy image.

  12. Methodology for diagnosing of skin cancer on images of dermatologic spots by spectral analysis.

    PubMed

    Guerra-Rosas, Esperanza; Álvarez-Borrego, Josué

    2015-10-01

    In this paper a new methodology for the diagnosing of skin cancer on images of dermatologic spots using image processing is presented. Currently skin cancer is one of the most frequent diseases in humans. This methodology is based on Fourier spectral analysis by using filters such as the classic, inverse and k-law nonlinear. The sample images were obtained by a medical specialist and a new spectral technique is developed to obtain a quantitative measurement of the complex pattern found in cancerous skin spots. Finally a spectral index is calculated to obtain a range of spectral indices defined for skin cancer. Our results show a confidence level of 95.4%.

  13. Spectral estimation of artist oil paints using multi-filter trichromatic imaging

    NASA Astrophysics Data System (ADS)

    Imai, Francisco H.; Berns, Roy S.

    2002-06-01

    A practical and easy way to capture images of oil-paintings and estimate their spectral reflectance as a function of position was tested. For the image acquisition, a trichromatic digital camera was used in conjunction with an absorption filter producing six channels. From an a priori statistical analysis of common artist oil paints, spectral reflectance was estimated. These experiments showed that it is possible to estimate the spectral reflectance with an accuracy of average ΔE*94 of 1.7 and spectral reflectance rms error of 2.2%. Of particular interest is guidance towards the design of a universal calibration target for imaging paintings.

  14. Multi-photon imaging of tumor cell invasion in an orthotopic mouse model of oral squamous cell carcinoma.

    PubMed

    Gatesman Ammer, Amanda; Hayes, Karen E; Martin, Karen H; Zhang, Lingqing; Spirou, George A; Weed, Scott A

    2011-07-25

    Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1(nu/nu) mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

  15. Forensic inspection of document using visible and near-infrared spectral imaging

    NASA Astrophysics Data System (ADS)

    Huang, Wei; Wang, Guiqiang; Xu, Xiaojing; Yu, Tao; Yang, Zhicheng

    2010-11-01

    In this study, the potential of visible and near-infrared spectral imaging as a technique of document inspection was examined. Doubtful documents are often found in economic cases, distinguished between original and added strokes and detected blurry characters are very useful for judgment. Burned, covered and rinsed documents in which the characters can't be identified with naked eyes were experimentally studied with a visible spectral imaging technique. Meanwhile, the same color inks were detected by both visible and near-infrared imaging spectrometers. Classification of spectral images was carried out in specialist spectral imaging software packager Misystem provided by Institute of Forensic Science. The technique significantly improved the detection of many documents, especially those that might be considered of poor quality or borderline characters. The visible spectral imaging was successful in detecting the burnt Chinese characters produced using pencils. It was possible to form spectral images showing the strokes even covered by Chinese ink by means of imaging at characteristic frequencies. As inks have very different spectral from the clothes, contribution and contrast of the rinsed lines and illegible seal words on clothes were clearly enhanced. By examining the spectral images from the inks, it was possible to determine whether the same color inks were written by the different pens. The results also show that the near-infrared spectrometer is better than visible one in distinguishing the same inks. In blind testing, spectral imaging was shown to achieve an average 85.1% chance of success. The results reveal the wide applications of spectral imaging in document evidence analysis. The potential of this technique in forensic science will be more apparent along with the further and deeper studies.

  16. Feature Transformation Detection Method with Best Spectral Band Selection Process for Hyper-spectral Imaging

    NASA Astrophysics Data System (ADS)

    Chen, Hai-Wen; McGurr, Mike; Brickhouse, Mark

    2015-11-01

    We present a newly developed feature transformation (FT) detection method for hyper-spectral imagery (HSI) sensors. In essence, the FT method, by transforming the original features (spectral bands) to a different feature domain, may considerably increase the statistical separation between the target and background probability density functions, and thus may significantly improve the target detection and identification performance, as evidenced by the test results in this paper. We show that by differentiating the original spectral, one can completely separate targets from the background using a single spectral band, leading to perfect detection results. In addition, we have proposed an automated best spectral band selection process with a double-threshold scheme that can rank the available spectral bands from the best to the worst for target detection. Finally, we have also proposed an automated cross-spectrum fusion process to further improve the detection performance in lower spectral range (<1000 nm) by selecting the best spectral band pair with multivariate analysis. Promising detection performance has been achieved using a small background material signature library for concept-proving, and has then been further evaluated and verified using a real background HSI scene collected by a HYDICE sensor.

  17. Spectral analysis for evaluation of myocardial tracers for medical imaging

    SciTech Connect

    Huesman, Ronald H.; Reutter, Bryan W.; Marshall, Robert C.

    2000-10-11

    Kinetic analysis of dynamic tracer data is performed with the goal of evaluating myocardial radiotracers for cardiac nuclear medicine imaging. Data from experiments utilizing the isolated rabbit heart model are acquired by sampling the venous blood after introduction of a tracer of interest and a reference tracer. We have taken the approach that the kinetics are properly characterized by an impulse response function which describes the difference between the reference molecule (which does not leave the vasculature) and the molecule of interest which is transported across the capillary boundary and is made available to the cell. Using this formalism we can model the appearance of the tracer of interest in the venous output of the heart as a convolution of the appearance of the reference tracer with the impulse response. In this work we parameterize the impulse response function as the sum of a large number of exponential functions whose predetermined decay constants form a spectrum, and each is required only to have a nonnegative coefficient. This approach, called spectral analysis, has the advantage that it allows conventional compartmental analysis without prior knowledge of the number of compartments which the physiology may require or which the data will support.

  18. Spectral Regression Discriminant Analysis for Hyperspectral Image Classification

    NASA Astrophysics Data System (ADS)

    Pan, Y.; Wu, J.; Huang, H.; Liu, J.

    2012-08-01

    Dimensionality reduction algorithms, which aim to select a small set of efficient and discriminant features, have attracted great attention for Hyperspectral Image Classification. The manifold learning methods are popular for dimensionality reduction, such as Locally Linear Embedding, Isomap, and Laplacian Eigenmap. However, a disadvantage of many manifold learning methods is that their computations usually involve eigen-decomposition of dense matrices which is expensive in both time and memory. In this paper, we introduce a new dimensionality reduction method, called Spectral Regression Discriminant Analysis (SRDA). SRDA casts the problem of learning an embedding function into a regression framework, which avoids eigen-decomposition of dense matrices. Also, with the regression based framework, different kinds of regularizes can be naturally incorporated into our algorithm which makes it more flexible. It can make efficient use of data points to discover the intrinsic discriminant structure in the data. Experimental results on Washington DC Mall and AVIRIS Indian Pines hyperspectral data sets demonstrate the effectiveness of the proposed method.

  19. Wide-Field Imaging Interferometry Spatial-Spectral Image Synthesis Algorithms

    NASA Technical Reports Server (NTRS)

    Lyon, Richard G.; Leisawitz, David T.; Rinehart, Stephen A.; Memarsadeghi, Nargess; Sinukoff, Evan J.

    2012-01-01

    Developed is an algorithmic approach for wide field of view interferometric spatial-spectral image synthesis. The data collected from the interferometer consists of a set of double-Fourier image data cubes, one cube per baseline. These cubes are each three-dimensional consisting of arrays of two-dimensional detector counts versus delay line position. For each baseline a moving delay line allows collection of a large set of interferograms over the 2D wide field detector grid; one sampled interferogram per detector pixel per baseline. This aggregate set of interferograms, is algorithmically processed to construct a single spatial-spectral cube with angular resolution approaching the ratio of the wavelength to longest baseline. The wide field imaging is accomplished by insuring that the range of motion of the delay line encompasses the zero optical path difference fringe for each detector pixel in the desired field-of-view. Each baseline cube is incoherent relative to all other baseline cubes and thus has only phase information relative to itself. This lost phase information is recovered by having point, or otherwise known, sources within the field-of-view. The reference source phase is known and utilized as a constraint to recover the coherent phase relation between the baseline cubes and is key to the image synthesis. Described will be the mathematical formalism, with phase referencing and results will be shown using data collected from NASA/GSFC Wide-Field Imaging Interferometry Testbed (WIIT).

  20. Quantitative method to assess caries via fluorescence imaging from the perspective of autofluorescence spectral analysis

    NASA Astrophysics Data System (ADS)

    Chen, Q. G.; Zhu, H. H.; Xu, Y.; Lin, B.; Chen, H.

    2015-08-01

    A quantitative method to discriminate caries lesions for a fluorescence imaging system is proposed in this paper. The autofluorescence spectral investigation of 39 teeth samples classified by the International Caries Detection and Assessment System levels was performed at 405 nm excitation. The major differences in the different caries lesions focused on the relative spectral intensity range of 565-750 nm. The spectral parameter, defined as the ratio of wavebands at 565-750 nm to the whole spectral range, was calculated. The image component ratio R/(G + B) of color components was statistically computed by considering the spectral parameters (e.g. autofluorescence, optical filter, and spectral sensitivity) in our fluorescence color imaging system. Results showed that the spectral parameter and image component ratio presented a linear relation. Therefore, the image component ratio was graded as <0.66, 0.66-1.06, 1.06-1.62, and >1.62 to quantitatively classify sound, early decay, established decay, and severe decay tissues, respectively. Finally, the fluorescence images of caries were experimentally obtained, and the corresponding image component ratio distribution was compared with the classification result. A method to determine the numerical grades of caries using a fluorescence imaging system was proposed. This method can be applied to similar imaging systems.

  1. Short-wave infrared (SWIR) spectral imager based on Fabry-Perot interferometer for remote sensing

    NASA Astrophysics Data System (ADS)

    Mannila, Rami; Holmlund, Christer; Ojanen, Harri J.; Näsilä, Antti; Saari, Heikki

    2014-10-01

    VTT Technical Research Centre of Finland has developed a spectral imager for short-wave infrared (SWIR) wavelength range. The spectral imager is based on a tunable Fabry-Perot interferometer (FPI) accompanied by a commercial InGaAs Camera. The FPI consists of two dielectric coated mirrors separated by a tunable air gap. Tuning of the air gap tunes also transmitted wavelength and therefore FPI acts as a tunable band bass filter. The FPI is piezo-actuated and it uses three piezo-actuators in a closed capacitive feedback loop for air gap tuning. The FPI has multiple order transmission bands, which limit free spectral range. Therefore spectral imager contains two FPI in a stack, to make possible to cover spectral range of 1000 - 1700 nm. However, in the first tests imager was used with one FPI and spectral range was limited to 1100-1600 nm. The spectral resolution of the imager is approximately 15 nm (FWHM). Field of view (FOV) across the flight direction is 30 deg. Imaging resolution of the spectral imager is 256 x 320 pixels. The focal length of the optics is 12 mm and F-number is 3.2. This imager was tested in summer 2014 in an unmanned aerial vehicle (UAV) and therefore a size and a mass of the imager were critical. Total mass of the imager is approximately 1200 grams. In test campaign the spectral imager will be used for forest and agricultural imaging. In future, because results of the UAV test flights are promising, this technology can be applied to satellite applications also.

  2. [The hyperspectral camera side-scan geometric imaging in any direction considering the spectral mixing].

    PubMed

    Wang, Shu-Min; Zhang, Ai-Wu; Hu, Shao-Xing; Sun, Wei-Dong

    2014-07-01

    In order to correct the image distortion in the hyperspectral camera side-scan geometric Imaging, the image pixel geo-referenced algorithm was deduced in detail in the present paper, which is suitable to the linear push-broom camera side-scan imaging on the ground in any direction. It takes the orientation of objects in the navigation coordinates system into account. Combined with the ground sampling distance of geo-referenced image and the area of push broom imaging, the general process of geo-referenced image divided into grids is also presented. The new image rows and columns will be got through the geo-referenced image area dividing the ground sampling distance. Considering the error produced by round rule in the pixel grids generated progress, and the spectral mixing problem caused by traditional direct spectral sampling method in the process of image correction, the improved spectral sampling method based on the weighted fusion method was proposed. It takes the area proportion of adjacent pixels in the new generated pixel as coefficient and then the coefficients are normalized to avoid the spectral overflow. So the new generated pixel is combined with the geo-referenced adjacent pixels spectral. Finally the amounts of push-broom imaging experiments were taken on the ground, and the distortion images were corrected according to the algorithm proposed above. The results show that the linear image distortion correction algorithm is valid and robust. At the same time, multiple samples were selected in the corrected images to verify the spectral data. The results indicate that the improved spectral sampling method is better than the direct spectral sampling algorithm. It provides reference for the application of similar productions on the ground. PMID:25269321

  3. Spectral images browsing using principal component analysis and set partitioning in hierarchical tree

    NASA Astrophysics Data System (ADS)

    Ma, Long; Zhao, Deping

    2011-12-01

    Spectral imaging technology have been used mostly in remote sensing, but have recently been extended to new area requiring high fidelity color reproductions like telemedicine, e-commerce, etc. These spectral imaging systems are important because they offer improved color reproduction quality not only for a standard observer under a particular illuminantion, but for any other individual exhibiting normal color vision capability under another illuminantion. A possibility for browsing of the archives is needed. In this paper, the authors present a new spectral image browsing architecture. The architecture for browsing is expressed as follow: (1) The spectral domain of the spectral image is reduced with the PCA transform. As a result of the PCA transform the eigenvectors and the eigenimages are obtained. (2) We quantize the eigenimages with the original bit depth of spectral image (e.g. if spectral image is originally 8bit, then quantize eigenimage to 8bit), and use 32bit floating numbers for the eigenvectors. (3) The first eigenimage is lossless compressed by JPEG-LS, the other eigenimages were lossy compressed by wavelet based SPIHT algorithm. For experimental evalution, the following measures were used. We used PSNR as the measurement for spectral accuracy. And for the evaluation of color reproducibility, ΔE was used.here standard D65 was used as a light source. To test the proposed method, we used FOREST and CORAL spectral image databases contrain 12 and 10 spectral images, respectively. The images were acquired in the range of 403-696nm. The size of the images were 128*128, the number of bands was 40 and the resolution was 8 bits per sample. Our experiments show the proposed compression method is suitable for browsing, i.e., for visual purpose.

  4. Compressive spectral polarization imaging by a pixelized polarizer and colored patterned detector.

    PubMed

    Fu, Chen; Arguello, Henry; Sadler, Brian M; Arce, Gonzalo R

    2015-11-01

    A compressive spectral and polarization imager based on a pixelized polarizer and colored patterned detector is presented. The proposed imager captures several dispersed compressive projections with spectral and polarization coding. Stokes parameter images at several wavelengths are reconstructed directly from 2D projections. Employing a pixelized polarizer and colored patterned detector enables compressive sensing over spatial, spectral, and polarization domains, reducing the total number of measurements. Compressive sensing codes are specially designed to enhance the peak signal-to-noise ratio in the reconstructed images. Experiments validate the architecture and reconstruction algorithms.

  5. High-resolution multimodal clinical multiphoton tomography of skin

    NASA Astrophysics Data System (ADS)

    König, Karsten

    2011-03-01

    This review focuses on multimodal multiphoton tomography based on near infrared femtosecond lasers. Clinical multiphoton tomographs for 3D high-resolution in vivo imaging have been placed into the market several years ago. The second generation of this Prism-Award winning High-Tech skin imaging tool (MPTflex) was introduced in 2010. The same year, the world's first clinical CARS studies have been performed with a hybrid multimodal multiphoton tomograph. In particular, non-fluorescent lipids and water as well as mitochondrial fluorescent NAD(P)H, fluorescent elastin, keratin, and melanin as well as SHG-active collagen has been imaged with submicron resolution in patients suffering from psoriasis. Further multimodal approaches include the combination of multiphoton tomographs with low-resolution wide-field systems such as ultrasound, optoacoustical, OCT, and dermoscopy systems. Multiphoton tomographs are currently employed in Australia, Japan, the US, and in several European countries for early diagnosis of skin cancer, optimization of treatment strategies, and cosmetic research including long-term testing of sunscreen nanoparticles as well as anti-aging products.

  6. Analysis of the metabolic deterioration of ex vivo skin from ischemic necrosis through the imaging of intracellular NAD(P)H by multiphoton tomography and fluorescence lifetime imaging microscopy

    NASA Astrophysics Data System (ADS)

    Sanchez, Washington Y.; Prow, Tarl W.; Sanchez, Washington H.; Grice, Jeffrey E.; Roberts, Michael S.

    2010-07-01

    Ex vivo human skin has been used extensively for cosmeceutical and drug delivery studies, transplantable skin allografts, or skin flaps. However, it has a half-life of a few days due to ischemic necrosis. Traditional methods of assessing viability can be time-consuming and provide limited metabolic information. Using multiphoton tomography and fluorescence lifetime imaging (MPT-FLIM) we assess ischemic necrosis of ex vivo skin by NAD(P)H autofluorescence intensity and fluorescence lifetime. Ex vivo skin is stored in the presence and absence of nutrient media (Dulbecco Modified Eagle Medium) at -20, 4, and 37 °C and room temperature over a 7-day time course to establish different rates of metabolic deterioration. At higher temperatures we observe a decrease in NAD(P)H autofluorescence, higher image noise, and a significant increase in the average fluorescence lifetime (τm) from ~1000 to 2000 ps. Additionally, significant distortions in NAD(P)H fluorescence lifetime histograms correspond to the reduction in autofluorescence. Skin kept at 4 °C, with or without media, showed the least change. Our findings suggest that MPT-FLIM enables useful noninvasive optical biopsies to monitor the metabolic state and deterioration of human skin for research and clinical purposes.

  7. The application of high spectral and spatial resolution imaging spectrometers for locating downed aircraft

    NASA Technical Reports Server (NTRS)

    Gatlin, James A.; Middleton, Elizabeth M.; Irons, James R.; Robinson, Jon W.

    1991-01-01

    The utility of high-resolution imaging spectrometer data is examined as an aid in locating downed aircraft by using a unique spectral signature while not requiring the extremely high spatial resolution needed to identify an aircraft by shape. Ground spectral measurements of several airplane wings, overflight spectral measurements of aircraft scenes, and the rationale for the chosen spectral signature are presented. It is concluded that imaging spectrometers which can detect and spatially locate a narrow-band spectral signature filling only a few pixels appear to have a utility for search and rescue aircraft or satellite systems as a aid in locating small downed aircraft. This spectral feature would have to be added to the surface coatings applied to aircraft. Proposed for use as such a spectral signature is a significant negative reflectance slope, in the 520 to 580 nm interval.

  8. Generalized Multiphoton Quantum Interference

    NASA Astrophysics Data System (ADS)

    Tillmann, Max; Tan, Si-Hui; Stoeckl, Sarah E.; Sanders, Barry C.; de Guise, Hubert; Heilmann, René; Nolte, Stefan; Szameit, Alexander; Walther, Philip

    2015-10-01

    Nonclassical interference of photons lies at the heart of optical quantum information processing. Here, we exploit tunable distinguishability to reveal the full spectrum of multiphoton nonclassical interference. We investigate this in theory and experiment by controlling the delay times of three photons injected into an integrated interferometric network. We derive the entire coincidence landscape and identify transition matrix immanants as ideally suited functions to describe the generalized case of input photons with arbitrary distinguishability. We introduce a compact description by utilizing a natural basis that decouples the input state from the interferometric network, thereby providing a useful tool for even larger photon numbers.

  9. Multi-photon excitation microscopy

    PubMed Central

    Diaspro, Alberto; Bianchini, Paolo; Vicidomini, Giuseppe; Faretta, Mario; Ramoino, Paola; Usai, Cesare

    2006-01-01

    Multi-photon excitation (MPE) microscopy plays a growing role among microscopical techniques utilized for studying biological matter. In conjunction with confocal microscopy it can be considered the imaging workhorse of life science laboratories. Its roots can be found in a fundamental work written by Maria Goeppert Mayer more than 70 years ago. Nowadays, 2PE and MPE microscopes are expected to increase their impact in areas such biotechnology, neurobiology, embryology, tissue engineering, materials science where imaging can be coupled to the possibility of using the microscopes in an active way, too. As well, 2PE implementations in noninvasive optical bioscopy or laser-based treatments point out to the relevance in clinical applications. Here we report about some basic aspects related to the phenomenon, implications in three-dimensional imaging microscopy, practical aspects related to design and realization of MPE microscopes, and we only give a list of potential applications and variations on the theme in order to offer a starting point for advancing new applications and developments. PMID:16756664

  10. Multi-photon excitation microscopy.

    PubMed

    Diaspro, Alberto; Bianchini, Paolo; Vicidomini, Giuseppe; Faretta, Mario; Ramoino, Paola; Usai, Cesare

    2006-01-01

    Multi-photon excitation (MPE) microscopy plays a growing role among microscopical techniques utilized for studying biological matter. In conjunction with confocal microscopy it can be considered the imaging workhorse of life science laboratories. Its roots can be found in a fundamental work written by Maria Goeppert Mayer more than 70 years ago. Nowadays, 2PE and MPE microscopes are expected to increase their impact in areas such biotechnology, neurobiology, embryology, tissue engineering, materials science where imaging can be coupled to the possibility of using the microscopes in an active way, too. As well, 2PE implementations in noninvasive optical bioscopy or laser-based treatments point out to the relevance in clinical applications. Here we report about some basic aspects related to the phenomenon, implications in three-dimensional imaging microscopy, practical aspects related to design and realization of MPE microscopes, and we only give a list of potential applications and variations on the theme in order to offer a starting point for advancing new applications and developments.

  11. Multi-photon excitation microscopy.

    PubMed

    Diaspro, Alberto; Bianchini, Paolo; Vicidomini, Giuseppe; Faretta, Mario; Ramoino, Paola; Usai, Cesare

    2006-01-01

    Multi-photon excitation (MPE) microscopy plays a growing role among microscopical techniques utilized for studying biological matter. In conjunction with confocal microscopy it can be considered the imaging workhorse of life science laboratories. Its roots can be found in a fundamental work written by Maria Goeppert Mayer more than 70 years ago. Nowadays, 2PE and MPE microscopes are expected to increase their impact in areas such biotechnology, neurobiology, embryology, tissue engineering, materials science where imaging can be coupled to the possibility of using the microscopes in an active way, too. As well, 2PE implementations in noninvasive optical bioscopy or laser-based treatments point out to the relevance in clinical applications. Here we report about some basic aspects related to the phenomenon, implications in three-dimensional imaging microscopy, practical aspects related to design and realization of MPE microscopes, and we only give a list of potential applications and variations on the theme in order to offer a starting point for advancing new applications and developments. PMID:16756664

  12. The characteristic analysis of spectral image for cabbage leaves damaged by diamondback moth pests

    NASA Astrophysics Data System (ADS)

    Lin, Li-bo; Li, Hong-ning; Cao, Peng-fei; Qin, Feng; Yang, Shu-ming; Feng, Jie

    2015-02-01

    Cabbage growth and health diagnosis are important parts for cabbage fine planting, spectral imaging technology with the advantages of obtaining spectrum and space information of the target at the same time, which has become a research hotspot at home and abroad. The experiment measures the reflection spectrum at different stages using liquid crystal tunable filter (LCTF) and monochromatic CMOS camera composed of spectral imaging system for cabbage leaves damaged by diamondback moth pests, and analyzes its feature bands and the change of spectral parameters. The study shows that the feature bands of cabbage leaves damaged by diamondback moth pests have a tendency to blue light direction, the red edge towards blue shift, and red valley raising in spectral characteristic parameters, which have a good indication in diagnosing the extent of cabbage damaged by pests. Therefore, it has a unique advantage of monitoring the cabbage leaves damaged by diamondback moth pests by combinating feature bands and spectral characteristic parameters in spectral imaging technology.

  13. Laser-induced fluorescence imaging of subsurface tissue structures with a volume holographic spatial-spectral imaging system.

    PubMed

    Luo, Yuan; Gelsinger-Austin, Paul J; Watson, Jonathan M; Barbastathis, George; Barton, Jennifer K; Kostuk, Raymond K

    2008-09-15

    A three-dimensional imaging system incorporating multiplexed holographic gratings to visualize fluorescence tissue structures is presented. Holographic gratings formed in volume recording materials such as a phenanthrenquinone poly(methyl methacrylate) photopolymer have narrowband angular and spectral transmittance filtering properties that enable obtaining spatial-spectral information within an object. We demonstrate this imaging system's ability to obtain multiple depth-resolved fluorescence images simultaneously.

  14. Laser-induced fluorescence imaging of subsurface tissue structures with a volume holographic spatial-spectral imaging system.

    PubMed

    Luo, Yuan; Gelsinger-Austin, Paul J; Watson, Jonathan M; Barbastathis, George; Barton, Jennifer K; Kostuk, Raymond K

    2008-09-15

    A three-dimensional imaging system incorporating multiplexed holographic gratings to visualize fluorescence tissue structures is presented. Holographic gratings formed in volume recording materials such as a phenanthrenquinone poly(methyl methacrylate) photopolymer have narrowband angular and spectral transmittance filtering properties that enable obtaining spatial-spectral information within an object. We demonstrate this imaging system's ability to obtain multiple depth-resolved fluorescence images simultaneously. PMID:18794943

  15. Liquid-crystal tunable filter spectral imaging for brain tumor demarcation

    NASA Astrophysics Data System (ADS)

    Gebhart, Steven C.; Thompson, Reid C.; Mahadevan-Jansen, Anita

    2007-04-01

    Past studies have demonstrated that combined fluorescence and diffuse reflectance spectroscopy can successfully discriminate between normal, tumor core, and tumor margin tissues in the brain. To achieve efficient, real-time surgical resection guidance with optical biopsy, probe-based spectroscopy must be extended to spectral imaging to spatially demarcate the tumor margins. We describe the design and characterization of a combined fluorescence and diffuse reflectance imaging system that uses liquid-crystal tunable filter technology. Experiments were conducted to quantitatively determine the linearity, field of view, spatial and spectral resolution, and wavelength sensitivity of the imaging system. Spectral images were acquired from tissue phantoms, mouse brain in vitro, and human cortex in vivo for functional testing of the system. The spectral imaging system produces measured intensities that are linear with sample emission intensity and integration time and possesses a 1 in. (2.54 cm) field of view for a 7 in. (18 cm) object distance. The spectral resolution is linear with wavelength, and the spatial resolution is pixel-limited. The sensitivity spectra for the imaging system provide a guide for the distribution of total image integration time between wavelengths. Functional tests in vitro demonstrate the capability to spectrally discriminate between brain tissues based on exogenous fluorescence contrast or endogenous tissue composition. In vivo imaging captures adequate fluorescence and diffuse reflectance intensities within a clinically viable 2 min imaging time frame and demonstrates the importance of hemostasis to acquired signal strengths and imaging speed.

  16. Design of multi-spectral images real-time segmentation system

    NASA Astrophysics Data System (ADS)

    Zhai, Bo; Qu, Youshan; Han, Yameng; Zhou, Jiang

    2015-02-01

    In order to realize the real-time segmentation processing of multi spectral images in practice, a real-time multi-spectral images segmentation system composed of four TMS320C6455 DSPs, two Virtex-4 - V4 XC4VLX80 - FPGAs and one Virtex-2 Pro - V2 Pro20 - FPGA is designed. Through the optimization of the cooperation processing of the multi DSP and multi FPGA, the parallel multitask processing ability of the DSPs and the effective interface coordination ability of the FPGAs in the built system are used fully. In order to display the processing ability, the segmentation test experiments of 10 spectra visible images, with 1024×1024, segmented by the Multi-scale Image Segmentation Method, was done in the built multi spectral images segment system. The experiment results prove that the multi DSP and multi FPGA multi spectral images processing system designed in this paper satisfies the real-time processing requirement in engineering practice.

  17. Fibre-coupled multiphoton microscope with adaptive motion compensation.

    PubMed

    Sherlock, Ben; Warren, Sean; Stone, James; Neil, Mark; Paterson, Carl; Knight, Jonathan; French, Paul; Dunsby, Chris

    2015-05-01

    To address the challenge of sample motion during in vivo imaging, we present a fibre-coupled multiphoton microscope with active axial motion compensation. The position of the sample surface is measured using optical coherence tomography and fed back to a piezo actuator that adjusts the axial location of the objective to compensate for sample motion. We characterise the system's performance and demonstrate that it can compensate for axial sample velocities up to 700 µm/s. Finally we illustrate the impact of motion compensation when imaging multiphoton excited autofluorescence in ex vivo mouse skin.

  18. Fibre-coupled multiphoton microscope with adaptive motion compensation

    PubMed Central

    Sherlock, Ben; Warren, Sean; Stone, James; Neil, Mark; Paterson, Carl; Knight, Jonathan; French, Paul; Dunsby, Chris

    2015-01-01

    To address the challenge of sample motion during in vivo imaging, we present a fibre-coupled multiphoton microscope with active axial motion compensation. The position of the sample surface is measured using optical coherence tomography and fed back to a piezo actuator that adjusts the axial location of the objective to compensate for sample motion. We characterise the system’s performance and demonstrate that it can compensate for axial sample velocities up to 700 µm/s. Finally we illustrate the impact of motion compensation when imaging multiphoton excited autofluorescence in ex vivo mouse skin. PMID:26137387

  19. Nonlinear dual-spectral image fusion for improving cone-beam-CT-based breast cancer diagnosis

    NASA Astrophysics Data System (ADS)

    Chen, Zikuan; Ning, Ruola; Conover, David; Willison, Kathleen

    2006-03-01

    Cone-beam breast computed tomography (CB Breast CT) can easily detect micro-calcifications and distinguish fat and glandular tissues from normal breast tissue. However, it may be a challenging task for CB Breast CT to distinguish benign from malignant tumors because of the subtle difference in x-ray attenuation. Due to the use of polyenergetic x-ray source, the x-ray and tissue interaction exhibits energy-dependent attenuation behavior, a phenomenon that, to date, has not been used for breast tissue characterization. We will exploit this spectral nature by equipping our CB Breast CT with dual-spectral imaging. The dual-spectral cone-beam scanning produces two spectral image datasets, from which we propose a nonlinear dual-spectral image fusion scheme to combine them into a single dataset, thereby incorporating the spectral information. In implementation, we will perform dual-spectral image fusion through a bi-variable polynomial that can be established by applying dual-spectral imaging to a reference material (with eight different thicknesses). From the fused dataset, we can reconstruct a volume, called a reference-equivalent volume or a fusion volume. By selecting the benign tissue as a reference material, we obtain a benign-equivalent volume. Likewise, we obtain a malignant-equivalent volume as well. In the pursuit of the discrimination of benign versus malignant tissues in a breast image, we perform intra-image as well as inter-image processing. The intra-image processing is an intensity transformation imposed only to a tomographic breast image itself, while the inter-image processing is exerted on two tomographic images extracted from two volumes. The nonlinear fusion scheme possesses these properties: 1) no noise magnification; 2) no feature dimensionality problem, and 3) drastic enhancement among specific features offered by nonlinear mapping. Its disadvantage lies in the possible misinterpretation resulting from nonlinear mapping.

  20. Compact static imaging spectrometer combining spectral zooming capability with a birefringent interferometer.

    PubMed

    Li, Jie; Zhu, Jingping; Qi, Chun; Zheng, Chuanlin; Gao, Bo; Zhang, Yunyao; Hou, Xun

    2013-04-22

    A compact static birefringent imaging spectrometer (BIS) with spectral zooming capability is presented. It based on two identical Wollaston prisms and has no slit. The most significant advantage of the BIS is that we can conveniently select spectral resolution to adapt to different application requirements and greatly reduce the size of the spectral image data for capturing, saving, transferring, and processing. Also, we show this configuration blend the advantage of a grating spectrometer and a Michelson interferometer: extremely compact, robust, wide free spectral range and very high throughput. PMID:23609723

  1. Development and characterization of non-resonant multiphoton photoacoustic spectroscopy (NMPPAS) for brain tumor margining

    NASA Astrophysics Data System (ADS)

    Dahal, Sudhir

    ) and healthy tissue with over 99% accuracy. NMPPAS spectral features showed evident differences between tumor and healthy tissues, and ratiometric analysis ensured that only a few wavelengths could be used for excitation instead of using numerous wavelength excitations to create spectra. This process would significantly reduce the analysis time while maintaining the same degree of accuracy. Tissue phantoms were fabricated in order to characterize the properties of NMPPAS. Scattering particles were doped into the phantoms to simulate their light scattering properties to real tissues. This allowed for better control over shape, size, reproducibility and doping in the sample while maintaining the light-tissue interaction properties of real tissue. To make NMPPAS viable for clinical applications, the technique was characterized to determine the spatial (lateral and longitudinal) resolution, depth of penetration and its ability to image in three-dimension through layers of tissue. Both resolutions were determined to be near-cellular level resolution (50-70 microm), obtained initially with the aid of the technique of multiphoton fluorescence, and later verified using NMPPAS imaging. Additionally, the maximum depth of penetration and detection was determined to be about 1.4cm, making the technique extremely suitable to margin tumors from underlying tissues in the brain. The capability of NMPPAS to detect and image layers that lie beneath other structures and blood vessels was also investigated. Three-dimensional images were obtained for the first time using NMPPAS. The images were obtained from different depths and structures were imaged through other layers of existing structures in the sample. This verified that NMPPAS was capable of detecting and imaging structures that lie embedded within the tissues. NMPPAS images of embedded structures were also obtained with the presence of hemoglobin, which is potentially the largest source of background in blood-perfused tissues

  2. Multiphoton harvesting metal–organic frameworks

    PubMed Central

    Quah, Hong Sheng; Chen, Weiqiang; Schreyer, Martin K.; Yang, Hui; Wong, Ming Wah; Ji, Wei; Vittal, Jagadese J.

    2015-01-01

    Multiphoton upconversion is a process where two or more photons are absorbed simultaneously to excite an electron to an excited state and, subsequently, the relaxation of electron gives rise to the emission of a photon with frequency greater than those of the absorbed photons. Materials possessing such property attracted attention due to applications in biological imaging, photodynamic therapy, three-dimensional optical data storage, frequency-upconverted lasing and optical power limiting. Here we report four-photon upconversion in metal–organic frameworks containing the ligand, trans, trans-9,10-bis(4-pyridylethenyl)anthracene. The ligand has a symmetrical acceptor–π–donor–π–acceptor structure and a singlet biradical electronic ground state, which boosted its multiphoton absorption cross-sections. We demonstrate that the upconversion efficiency can be enhanced by Förster resonance energy transfer within host–guest metal–organic frameworks consisting of encapsulated high quantum yielding guest molecules. Using these strategies, metal–organic framework materials, which can exhibit frequency-upconverted photoluminescence excited by simultaneous multiphoton absorption, can be rationally designed and synthesized. PMID:26245741

  3. Multiphoton harvesting metal-organic frameworks

    NASA Astrophysics Data System (ADS)

    Quah, Hong Sheng; Chen, Weiqiang; Schreyer, Martin K.; Yang, Hui; Wong, Ming Wah; Ji, Wei; Vittal, Jagadese J.

    2015-08-01

    Multiphoton upconversion is a process where two or more photons are absorbed simultaneously to excite an electron to an excited state and, subsequently, the relaxation of electron gives rise to the emission of a photon with frequency greater than those of the absorbed photons. Materials possessing such property attracted attention due to applications in biological imaging, photodynamic therapy, three-dimensional optical data storage, frequency-upconverted lasing and optical power limiting. Here we report four-photon upconversion in metal-organic frameworks containing the ligand, trans, trans-9,10-bis(4-pyridylethenyl)anthracene. The ligand has a symmetrical acceptor-π-donor-π-acceptor structure and a singlet biradical electronic ground state, which boosted its multiphoton absorption cross-sections. We demonstrate that the upconversion efficiency can be enhanced by Förster resonance energy transfer within host-guest metal-organic frameworks consisting of encapsulated high quantum yielding guest molecules. Using these strategies, metal-organic framework materials, which can exhibit frequency-upconverted photoluminescence excited by simultaneous multiphoton absorption, can be rationally designed and synthesized.

  4. Spectral CT imaging in patients with Budd-Chiari syndrome: investigation of image quality.

    PubMed

    Su, Lei; Dong, Junqiang; Sun, Qiang; Liu, Jie; Lv, Peijie; Hu, Lili; Yan, Liangliang; Gao, Jianbo

    2014-11-01

    To assess the image quality of monochromatic imaging from spectral CT in patients with Budd-Chiari syndrome (BCS), fifty patients with BCS underwent spectral CT to generate conventional 140 kVp polychromatic images (group A) and monochromatic images, with energy levels from 40 to 80, 40 + 70, and 50 + 70 keV fusion images (group B) during the portal venous phase (PVP) and the hepatic venous phase (HVP). Two-sample t tests compared vessel-to-liver contrast-to-noise ratio (CNR) and signal-to-noise ratio (SNR) for the portal vein (PV), hepatic vein (HV), inferior vena cava. Readers' subjective evaluations of the image quality were recorded. The highest SNR values in group B were distributed at 50 keV; the highest CNR values in group B were distributed at 40 keV. The higher CNR values and SNR values were obtained though PVP of PV (SNR 18.39 ± 6.13 vs. 10.56 ± 3.31, CNR 7.81 ± 3.40 vs. 3.58 ± 1.31) and HVP of HV (3.89 ± 2.08 vs. 1.27 ± 1.55) in the group B; the lower image noise for group B was at 70 keV and 50 + 70 keV (15.54 ± 8.39 vs. 18.40 ± 4.97, P = 0.0004 and 18.97 ± 7.61 vs. 18.40 ± 4.97, P = 0.0691); the results show that the 50 + 70 keV fusion image quality was better than that in group A. Monochromatic energy levels of 40-70, 40 + 70, and 50 + 70 keV fusion image can increase vascular contrast and that will be helpful for the diagnosis of BCS, we select the 50 + 70 keV fusion image to acquire the best BCS images.

  5. Multimodal optoacoustic and multiphoton fluorescence microscopy

    NASA Astrophysics Data System (ADS)

    Sela, Gali; Razansky, Daniel; Shoham, Shy

    2013-03-01

    Multiphoton microscopy is a powerful imaging modality that enables structural and functional imaging with cellular and sub-cellular resolution, deep within biological tissues. Yet, its main contrast mechanism relies on extrinsically administered fluorescent indicators. Here we developed a system for simultaneous multimodal optoacoustic and multiphoton fluorescence 3D imaging, which attains both absorption and fluorescence-based contrast by integrating an ultrasonic transducer into a two-photon laser scanning microscope. The system is readily shown to enable acquisition of multimodal microscopic images of fluorescently labeled targets and cell cultures as well as intrinsic absorption-based images of pigmented biological tissue. During initial experiments, it was further observed that that detected optoacoustically-induced response contains low frequency signal variations, presumably due to cavitation-mediated signal generation by the high repetition rate (80MHz) near IR femtosecond laser. The multimodal system may provide complementary structural and functional information to the fluorescently labeled tissue, by superimposing optoacoustic images of intrinsic tissue chromophores, such as melanin deposits, pigmentation, and hemoglobin or other extrinsic particle or dye-based markers highly absorptive in the NIR spectrum.

  6. [Influence of human body target's spectral characteristics on visual range of low light level image intensifiers].

    PubMed

    Zhang, Jun-Ju; Yang, Wen-Bin; Xu, Hui; Liu, Lei; Tao, Yuan-Yaun

    2013-11-01

    To study the effect of different human target's spectral reflective characteristic on low light level (LLL) image intensifier's distance, based on the spectral characteristics of the night-sky radiation and the spectral reflective coefficients of common clothes, we established a equation of human body target's spectral reflective distribution, and analyzed the spectral reflective characteristics of different human targets wearing the clothes of different color and different material, and from the actual detection equation of LLL image intensifier distance, discussed the detection capability of LLL image intensifier for different human target. The study shows that the effect of different human target's spectral reflective characteristic on LLL image intensifier distance is mainly reflected in the average reflectivity rho(-) and the initial contrast of the target and the background C0. Reflective coefficient and spectral reflection intensity of cotton clothes are higher than polyester clothes, and detection capability of LLL image intensifier is stronger for the human target wearing cotton clothes. Experimental results show that the LLL image intensifiers have longer visual ranges for targets who wear cotton clothes than targets who wear same color but polyester clothes, and have longer visual ranges for targets who wear light-colored clothes than targets who wear dark-colored clothes. And in the full moon illumination conditions, LLL image intensifiers are more sensitive to the clothes' material.

  7. Real-time dispersion-compensated image reconstruction for compressive sensing spectral domain optical coherence tomography.

    PubMed

    Xu, Daguang; Huang, Yong; Kang, Jin U

    2014-09-01

    In this work, we propose a novel dispersion compensation method that enables real-time compressive sensing (CS) spectral domain optical coherence tomography (SD OCT) image reconstruction. We show that dispersion compensation can be incorporated into CS SD OCT by multiplying the dispersion-correcting terms by the undersampled spectral data before CS reconstruction. High-quality SD OCT imaging with dispersion compensation was demonstrated at a speed in excess of 70 frames per s using 40% of the spectral measurements required by the well-known Shannon/Nyquist theory. The data processing and image display were performed on a conventional workstation having three graphics processing units.

  8. Initial clinical testing of a multi-spectral imaging system built on a smartphone platform

    NASA Astrophysics Data System (ADS)

    Mink, Jonah W.; Wexler, Shraga; Bolton, Frank J.; Hummel, Charles; Kahn, Bruce S.; Levitz, David

    2016-03-01

    Multi-spectral imaging systems are often expensive and bulky. An innovative multi-spectral imaging system was fitted onto a mobile colposcope, an imaging system built around a smartphone in order to image the uterine cervix from outside the body. The multi-spectral mobile colposcope (MSMC) acquires images at different wavelengths. This paper presents the clinical testing of MSMC imaging (technical validation of the MSMC system is described elsewhere 1 ). Patients who were referred to colposcopy following abnormal screening test (Pap or HPV DNA test) according to the standard of care were enrolled. Multi-spectral image sets of the cervix were acquired, consisting of images from the various wavelengths. Image acquisition took 1-2 sec. Areas suspected for dysplasia under white light imaging were biopsied, according to the standard of care. Biopsied sites were recorded on a clockface map of the cervix. Following the procedure, MSMC data was processed from the sites of biopsied sites. To date, the initial histopathological results are still outstanding. Qualitatively, structures in the cervical images were sharper at lower wavelengths than higher wavelengths. Patients tolerated imaging well. The result suggests MSMC holds promise for cervical imaging.

  9. New confocal microscopy hyperspectral imager for NIR-emitting bioprobes: high spectral resolution for a wide spectral range (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Marcet, Stéphane; Benayas, Antonio; Quintanilla, Marta; Mangiarini, Francesca; Verhaegen, Marc; Vetrone, Fiorenzo; Blais-Ouellette, Sébastien

    2016-03-01

    Functional nanoscale materials are being extensively investigated for applications in biology and medicine and are ready to make significant contributions in the realization of exciting advancements in diverse areas of diagnostics and therapeutics. Aiming for more accurate, efficient, non-invasive and fast diagnostic tools, the use of near-infrared (NIR) light in the range of the 1st and 2nd biological window (NIR-I: 0.70-0.95 µm; NIR-II: 1.00-1.35 µm) provides deeper penetration depth into biological tissue, better image contrast, reduced phototoxicity and photobleaching. Consequently, NIR-based bioimaging became a quickly emerging field and manifold new NIR-emitting bioprobes have been reported. Since commercially available microscopes are not optimized for this kind of NPs, a new microscopy hyperspectral confocal imager has been developed to cover a broad spectral range (400 to 1700 nm) with high spectral resolution. The smallest spectral variation can be easily monitored thanks to the high spectral resolution (as low as 0.2 nm). This is possible thanks to a combination of an EMCCD and an InGaAs camera with a high resolution spectrometer. An extended number of NPs can be excited with a Ti:Sapphire laser, which provides tunable illumination within 690-1040 nm. Cells and tissues can be mapped in less than 100 ms, allowing in-vivo imaging. As a proof of concept, here we present the preliminary results of the spatial distribution of the fluorescence signal intensity from lanthanide doped nanoparticles incorporated into a system of biological interest. The temperature sub-mm gradient - analyzing the spectral features so gathered through an all-optical route is also thoroughly discussed.

  10. Picosecond spectral coherent anti-Stokes Raman scattering imaging with principal component analysis of meibomian glands

    NASA Astrophysics Data System (ADS)

    Lin, Chia-Yu; Suhalim, Jeffrey L.; Nien, Chyong Ly; Miljković, Miloš D.; Diem, Max; Jester, James V.; Potma, Eric. O.

    2011-02-01

    The lipid distribution in the mouse meibomian gland was examined with picosecond spectral anti-Stokes Raman scattering (CARS) imaging. Spectral CARS data sets were generated by imaging specific localized regions of the gland within tissue sections at consecutive Raman shifts in the CH2 stretching vibrational range. Spectral differences between the location specific CARS spectra obtained in the lipid-rich regions of the acinus and the central duct were observed, which were confirmed with a Raman microspectroscopic analysis, and attributed to meibum lipid modifications within the gland. A principal component analysis of the spectral data set reveals changes in the CARS spectrum when transitioning from the acini to the central duct. These results demonstrate the utility of picosecond spectral CARS imaging combined with multivariate analysis for assessing differences in the distribution and composition of lipids in tissues.

  11. Spectral CT imaging of vulnerable plaque with two independent biomarkers

    NASA Astrophysics Data System (ADS)

    Baturin, Pavlo; Alivov, Yahya; Molloi, Sabee

    2012-07-01

    The purpose of this paper is to investigate the feasibility of a novel four-material decomposition technique for assessing the vulnerability of plaque with two contrast materials spectral computer tomography (CT) using two independent markers: plaque's inflammation and spotty calcification. A simulation study was conducted using an energy-sensitive photon-counting detector for k-edge imaging of the coronary arteries. In addition to detecting the inflammation status, which is known as a biological marker of a plaque's vulnerability, we use spotty calcium concentration as an independent marker to test a plaque's vulnerability. We have introduced a new method for detecting and quantifying calcium concentrations in the presence of two contrast materials (iodine and gold), calcium and soft tissue background. In this method, four-material decomposition was performed on a pixel-by-pixel basis, assuming there was an arbitrary mixture of materials in the voxel. The concentrations of iodine and gold were determined by the k-edge material decomposition based on the maximum likelihood method. The calibration curves of the attenuation coefficients, with respect to the concentrations of different materials, were used to separate the calcium signal from both contrast materials and different soft tissues in the mixtures. Three different materials (muscle, blood and lipid) were independently used as soft tissue. The simulations included both ideal and more realistic energy resolving detectors to measure the polychromatic photon spectrum in single slice parallel beam geometry. The ideal detector was used together with a 3 cm diameter digital phantom to demonstrate the decomposition method while a more realistic detector and a 33 × 24 cm2 digital chest phantom were simulated to validate the vulnerability assessment technique. A 120 kVp spectrum was generated to produce photon flux sufficient for detecting contrast materials above the k-edges of iodine (33.2 keV) and gold (80.7 ke

  12. Brain tumor demarcation with liquid-crystal tunable filter spectral imaging

    NASA Astrophysics Data System (ADS)

    Gebhart, Steven C.; Mahadevan-Jansen, Anita

    2006-02-01

    Past studies have demonstrated that combined fluorescence and diffuse reflectance spectroscopy can successfully discriminate between normal, tumor core, and tumor margin tissues in the brain. To achieve efficient surgical resection guidance with optical biopsy, probe-based spectroscopy must be extended to spectral imaging to spatially demarcate the tumor margins. This paper describes the design and testing of a combined fluorescence and diffuse reflectance imaging system which uses liquid-crystal tunable filter technology. Experiments were conducted to quantitatively determine its linearity, field of view, spatial and spectral resolution, and wavelength sensitivity. For functional testing, spectral images were acquired from tissue phantoms, mouse brain in vitro, and rat brain cortex in vivo. The spectral imaging system is characterized by measured intensities which are linear with sample emission intensity and integration time, a one-inch field of view for a seven-inch object distance, spectral resolution which is linear with wavelength, spatial resolution which is pixel-limited, and sensitivity functions which provide a guide for the distribution of total image integration time between wavelengths. Functional testing demonstrated good spatial and spectral constrast between brain tissue types, the capability to acquire adequate fluorescence and diffuse reflectance intensities within a one-minute imaging timeframe, and the importance of hemostasis to acquired signal strengths and imaging speed.

  13. Multiphoton ionization of Uracil

    NASA Astrophysics Data System (ADS)

    Prieto, Eladio; Martinez, Denhi; Guerrero, Alfonso; Alvarez, Ignacio; Cisneros, Carmen

    2016-05-01

    Multiphoton ionization and dissociation of Uracil using a Reflectron time of flight spectrometer was performed along with radiation from the second harmonic of a Nd:YAG laser. Uracil is one of the four nitrogen bases that belong to RNA. The last years special interest has been concentrated on the study of the effects under UV radiation in nucleic acids1 and also in the role that this molecule could have played in the origin and development of life on our planet.2 The MPI mass spectra show that the presence and intensity of the resulting ions strongly depend on the density power. The identification of the ions in the mass spectra is presented. The results are compared with those obtained in other laboratories under different experimental conditions and some of them show partial agreement.3 The present work was supported by CONACYT-Mexico Grant 165410 and DGAPA UNAM Grant IN101215 and IN102613.

  14. Spectral super-resolution reflectance retrieval from remotely sensed imaging spectrometer data.

    PubMed

    Jia, Guorui; Hueni, Andreas; Tao, Dongxing; Geng, Ruonan; Schaepman, Michael E; Zhao, Huijie

    2016-08-22

    Existing atmospheric correction methods retrieve surface reflectance keeping the same nominal spectral response functions (SRFs) as that of the airborne/spaceborne imaging spectrometer radiance data. Since the SRFs vary dependent on sensor type and configuration, the retrieved reflectance of the same ground object varies from sensor to sensor as well. This imposes evident limitations on data validation efforts between sensors at surface reflectance level. We propose a method to retrieve super-resolution reflectance at the surface, by combining the first-principles atmospheric correction method FLAASH (fast line-of-sight atmospheric analysis of spectral hypercubes) with spectral super-resolution of imaging spectrometer radiance data. This approach is validated by comparing airborne AVIRIS (airborne visible/infrared imaging spectrometer) and spaceborne Hyperion data. The results demonstrate that the super-resolution reflectance in spectral bands with sufficiently high signal-to-noise ratio (SNR) serves as intermediate quantity to cross validate data originating from different imaging spectrometers.

  15. RELIABILITY OF CONFOCAL MICROSCOPY SPECTRAL IMAGING SYSTEMS: USE OF MULTISPECTRAL BEADS

    EPA Science Inventory

    Background: There is a need for a standardized, impartial calibration, and validation protocol on confocal spectral imaging (CSI) microscope systems. To achieve this goal, it is necessary to have testing tools to provide a reproducible way to evaluate instrument performance. ...

  16. Intelligent spectral signature bio-imaging in vivo for surgical applications

    NASA Astrophysics Data System (ADS)

    Jeong, Jihoon; Frykman, Philip K.; Gaon, Mark; Chung, Alice P.; Lindsley, Erik H.; Hwang, Jae Y.; Farkas, Daniel L.

    2007-02-01

    Multi-spectral imaging provides digital images of a scene or object at a large, usually sequential number of wavelengths, generating precise optical spectra at every pixel. We use the term "spectral signature" for a quantitative plot of optical property variations as a function of wavelengths. We present here intelligent spectral signature bio-imaging methods we developed, including automatic signature selection based on machine learning algorithms and database search-based automatic color allocations, and selected visualization schemes matching these approaches. Using this intelligent spectral signature bio-imaging method, we could discriminate normal and aganglionic colon tissue of the Hirschsprung's disease mouse model with over 95% sensitivity and specificity in various similarity measure methods and various anatomic organs such as parathyroid gland, thyroid gland and pre-tracheal fat in dissected neck of the rat in vivo.

  17. Multispectral endoscopy and microscopy imaging system using a spectrally programmable light engine

    NASA Astrophysics Data System (ADS)

    MacKinnon, N.; Stange, Ulrich; Lane, Pierre M.; MacAulay, Calum E.

    2005-03-01

    We report a spectrally and temporally programmable light engine based on a spatial light modulator that can dynamically create any narrow or broadband spectral profile for hyperspectral, fluorescence, or principal component imaging. Most hyperspectral or multispectral imaging systems use wavelength selection devices such as acousto-optic tunable filters (AOTFs), tunable grating or prism-based monochromators, or filter wheels. While these devices can select wavelengths they cannot create arbitrary spectral profiles. This simple and economical system can be controlled at high speed (up to 5000 illumination profiles per second). Digitally controlled illumination is bit additive with image data providing high dynamic range imaging with monochrome or color imaging devices. This is especially advantageous for endoscopes employing small well CCD or CMOS sensors since the dynamic range now can extend beyond the limits of the sensor itself. In this report we show multispectral images of in vivo tissue and in vitro tissue samples using endoscopes, surgical microscopes and conventional microscopes.

  18. Detection of blood oxygen level by noninvasive passive spectral imaging of skin

    NASA Astrophysics Data System (ADS)

    Gupta, Neelam; Ramella-Roman, Jessica C.

    2008-02-01

    A compact optical hyperspectral imager that can detect both spectral and polarization signatures was used for passive noninvasive imaging of human skin. This vibration-insensitive imager uses an acousto-optic tunable filter (AOTF) as a spectral selection element and an electronically tunable liquid crystal variable retarder (LCVR) as a polarization device. Such an imager is ideally suited to provide both agile spectral and polarization signatures and can be readily used for real time in vivo medical imaging applications. Operation of this imager and image acquisition is fully computer controlled. This imager covers visible to near-infrared (VNIR) region from 400 to 800 nm with a 10 nm spectral resolution at 600 nm and uses a TeO II AOTF with a 15×15 mm2 linear aperture and a 4.2° angular aperture. At each wavelength 640×480 images with two orthogonal polarization are captured and a total of 41 spectral images are collected to form an image cube. A commercial Si CCD camera was used along with off-the-shelf lenses, mirrors and irises. We carried out experiments with a human subject and controlled the blood perfusion in the individual arm and finger by using a pressure cuff and a rubber band, respectively. Images were captured by illuminating the subject with a white light lamp source and imaging it from a distance. When the hyperspectral image analysis was performed we could observe the effects of skin deoxygenation. In this paper we will described our instrument, the experimental setup, the images obtained and the analysis results.

  19. Optical imaging of hemoglobin oxygen saturation using a small number of spectral images for endoscopic application

    NASA Astrophysics Data System (ADS)

    Saito, Takaaki; Yamaguchi, Hiroshi

    2015-12-01

    Tissue hypoxia is associated with tumor and inflammatory diseases, and detection of hypoxia is potentially useful for their detailed diagnosis. An endoscope system that can optically observe hemoglobin oxygen saturation (StO2) would enable minimally invasive, real-time detection of lesion hypoxia in vivo. Currently, point measurement of tissue StO2 via endoscopy is possible using the commercial fiber-optic oximeter T-Stat, which is based on visible light spectroscopy at many wavelengths. For clinical use, however, imaging of StO2 is desirable to assess the distribution of tissue oxygenation around a lesion. Here, we describe our StO2 imaging technique based on a small number of wavelength ranges in the visible range. By assuming a homogeneous tissue, we demonstrated that tissue StO2 can be obtained independently from the scattering property and blood concentration of tissue using four spectral bands. We developed a prototype endoscope system and used it to observe tissue-simulating phantoms. The StO2 (%) values obtained using our technique agreed with those from the T-Stat within 10%. We also showed that tissue StO2 can be derived using three spectral band if the scattering property is fixed at preliminarily measured values.

  20. Comparison of Spectral and Image Morphological Analysis for Egg Early Hatching Property Detection Based on Hyperspectral Imaging

    PubMed Central

    Zhang, Wei; Pan, Leiqing; Tu, Kang; Zhang, Qiang; Liu, Ming

    2014-01-01

    The use of non-destructive methods to detect egg hatching properties could increase efficiency in commercial hatcheries by saving space, reducing costs, and ensuring hatching quality. For this purpose, a hyperspectral imaging system was built to detect embryo development and vitality using spectral and morphological information of hatching eggs. A total of 150 green shell eggs were used, and hyperspectral images were collected for every egg on day 0, 1, 2, 3 and 4 of incubation. After imaging, two analysis methods were developed to extract egg hatching characteristic. Firstly, hyperspectral images of samples were evaluated using Principal Component Analysis (PCA) and only one optimal band with 822 nm was selected for extracting spectral characteristics of hatching egg. Secondly, an image segmentation algorithm was applied to isolate the image morphologic characteristics of hatching egg. To investigate the applicability of spectral and image morphological analysis for detecting egg early hatching properties, Learning Vector Quantization neural network (LVQNN) was employed. The experimental results demonstrated that model using image morphological characteristics could achieve better accuracy and generalization than using spectral characteristic parameters, and the discrimination accuracy for eggs with embryo development were 97% at day 3, 100% at day 4. In addition, the recognition results for eggs with weak embryo development reached 81% at day 3, and 92% at day 4. This study suggested that image morphological analysis was a novel application of hyperspectral imaging technology to detect egg early hatching properties. PMID:24551130

  1. New insights and system designs for temporally focused multiphoton optogenetics

    NASA Astrophysics Data System (ADS)

    Mayblum, Tom; Schejter, Adi; Dana, Hod; Shoham, Shy

    2015-03-01

    Temporal focusing (TF) multiphoton systems constitute a powerful solution for cellular resolution optogenetic stimulation and recording in three-dimensional, scattering tissue. Here, we address two fundamental aspects in the design of such systems: first, we examine the design of TF systems with specific optical sectioning by comparatively analyzing previously published results. Next, we develop a solution for obtaining TF in a flexible three-dimensional pattern of cellmatched focal spots. Our solution employs spatio-temporal focusing (SSTF) in a unique optical system design that can be integrated before essentially any multiphoton imaging or stimulation system.

  2. On combining spectral and spatial information of hyperspectral image for camouflaged target detecting

    NASA Astrophysics Data System (ADS)

    Hua, Wenshen; Liu, Xun; Yang, Jia

    2013-12-01

    Detecting enemy's targets and being undetectable play increasingly important roles in modern warfare. Hyperspectral images can provide large spectral range and high spectral resolution, which are invaluable in discriminating between camouflaged targets and backgrounds. As supervised classification requires prior knowledge which cannot be acquired easily, unsupervised classification usually is adopted to process hyperspectral images to detect camouflaged target. But one of its drawbacks—low detecting accuracy confines its application for camouflaged target detecting. Most research on the processing of hyperspectral image tends to focus exclusively on spectral domain and ignores spatial domain. However current hyperspectral image provides high spatial resolution which contains useful information for camouflaged target detecting. A new method combining spectral and spatial information is proposed to increase the detecting accuracy using unsupervised classification. The method has two steps. In the first step, a traditional unsupervised classifier (i.e. K-MEANS, ISODATA) is adopted to classify the hyperspectral image to acquire basic classifications or clusters. During the second step, a 3×3 model and spectral angle mapping are utilized to test the spatial character of the hyperspectral image. The spatial character is defined as spatial homogeneity and calculated by spectral angle mapping. Theory analysis and experiment shows the method is reasonable and efficient. Camouflaged targets are extracted from the background and different camouflaged targets are also recognized. And the proposed algorithm outperforms K-MEANS in terms of detecting accuracy, robustness and edge's distinction. This paper demonstrates the new method is meaningful to camouflaged targets detecting.

  3. Implementation of a multi-spectral color imaging device without color filter array

    NASA Astrophysics Data System (ADS)

    Langfelder, G.; Longoni, A. F.; Zaraga, F.

    2011-01-01

    In this work the use of the Transverse Field Detector (TFD) as a device for multispectral image acquisition is proposed. The TFD is a color imaging pixel capable of color reconstruction without color filters. Its basic working principle is based on the generation of a suitable electric field configuration inside a Silicon depleted region by means of biasing voltages applied to surface contacts. With respect to previously proposed methods for performing multispectral capture, the TFD has a unique characteristic of electrically tunable spectral responses. This feature allows capturing an image with different sets of spectral responses (RGB, R'G'B', and so on) simply by tuning the device biasing voltages in multiple captures. In this way no hardware complexity (no external filter wheels or varying sources) is added with respect to a colorimetric device. The estimation of the spectral reflectance of the area imaged by a TFD pixel is based in this work on a linear combination of six eigenfunctions. It is shown that a spectral reconstruction can be obtained either (1) using two subsequent image captures that generate six TFD spectral responses or (2) using a new asymmetric biasing scheme, which allows the implementation of five spectral responses for each TFD pixel site in a single configuration, definitely allowing one-shot multispectral imaging.

  4. Biomedical applications of the information-efficient spectral imaging sensor (ISIS)

    NASA Astrophysics Data System (ADS)

    Gentry, Stephen M.; Levenson, Richard M.

    1999-04-01

    The ISIS approach to spectral imaging seeks to bridge the gap between tuned multispectral and fixed hyperspectral imaging sensor. By allowing the definition of completely general spectral filter functions, truly optimal measurement can be made for a given task. These optimal measurements significantly improve signal to noise ratio and speed, minimize data volume and data rate, while preserving classification accuracy. This paper investigates the application of the ISIS sensing approach in two sample biomedical applications: prostate and colon cancer screening. It is shown that is these applications, two to three optimal measurements are sufficient to capture the majority of classification information for critical samples constituents. In the prostate cancer example, the optimal measurements allow 8 percent relative improvement in classification accuracy of critical cell constituents over a red, green, blue (RGB) sensor. In the colon cancer example, use of optimal measurements boost the classification accuracy of critical cell constituents by 28 percent relative to the RGB sensor. In both cases, optimal measurements match the performance achieved by the entire hyperspectral data set. The paper concludes that an ISIS style spectral imager can acquire these optimal spectral images directly, allowing improved classification accuracy imager can acquire these optimal spectral images directly, allowing improved classification accuracy over an RGB sensor. Compared to a hyperspectral sensor, the ISIS approach can achieve similar classification accuracy using a significantly lower number of spectral samples, thus minimizing overall sample classification time and cost.

  5. A novel and compact spectral imaging system based on two curved prisms

    NASA Astrophysics Data System (ADS)

    Nie, Yunfeng; Bin, Xiangli; Zhou, Jinsong; Li, Yang

    2013-09-01

    As a novel detection approach which simultaneously acquires two-dimensional visual picture and one-dimensional spectral information, spectral imaging offers promising applications on biomedical imaging, conservation and identification of artworks, surveillance of food safety, and so forth. A novel moderate-resolution spectral imaging system consisting of merely two optical elements is illustrated in this paper. It can realize the function of a relay imaging system as well as a 10nm spectral resolution spectroscopy. Compared to conventional prismatic imaging spectrometers, this design is compact and concise with only two special curved prisms by utilizing two reflective surfaces. In contrast to spectral imagers based on diffractive grating, the usage of compound-prism possesses characteristics of higher energy utilization and wider free spectral range. The seidel aberration theory and dispersive principle of this special prism are analyzed at first. According to the results, the optical system of this design is simulated, and the performance evaluation including spot diagram, MTF and distortion, is presented. In the end, considering the difficulty and particularity of manufacture and alignment, an available method for fabrication and measurement is proposed.

  6. Subsurface imaging using the spectral polarization difference technique and NIR illumination

    SciTech Connect

    Alfano, R R; Demos, S G; Radousky, H B

    1999-01-26

    A subsurface imaging system is utilized to test the ability of the spectral polarization difference imaging technique for deep subsurface imaging in tissues. The illumination of the system is derived from compact class III lasers in the red and NIR spectral region and, alternatively, from a white light source and selection of the appropriate illumination wavelength using band-pass optical filters. The experimental results demonstrate detection and imaging of a high-scattering object located up to 1.5 cm underneath the surface of a host chicken tissue.

  7. Multispectral imaging of papyri: area segregation by evaluation of their spectral signature correlation

    NASA Astrophysics Data System (ADS)

    Kovalchuk, Alexander

    2009-10-01

    Multispectral imaging technology was applied to deteriorated ancient manuscripts. Monochrome images were registered at twelve wavelengths. An image processing algorithm that utilises relative spectral intensity distribution for the areas of the surface (rather than absolute values of intensity of the reflected light) was evaluated. It was found that the areas in a cubed MSI image representing ink and papyrus were separable due to their distinctive spectral signatures. The developed technique for enhancing the contrast of the inscriptions was shown to work effectively over a large range of lightness for both ink and papyrus surface.

  8. Model-based restoration of underwater spectral images captured with narrowband filters.

    PubMed

    Guo, Yilu; Song, Hong; Liu, Hongbo; Wei, He; Yang, Ping; Zhan, Shuyue; Wang, Hangzhou; Huang, Hui; Liao, Ningfang; Mu, Quanquan; Leng, Jianxing; Yang, Wenjing

    2016-06-13

    Underwater spectral imaging is a promising method for mapping, classification and health monitoring of coral reefs and seafloor inhabitants. However, the spectrum of light is distorted during the underwater imaging process due to wavelength-dependent attenuation by the water. This paper presents a model-based method that accurately restores brightness of underwater spectral images captured with narrowband filters. A model is built for narrowband underwater spectral imaging. The model structure is derived from physical principles, representing the absorption, scattering and refraction by water and the optical properties of narrowband filters, lenses and image sensors. The model coefficients are calibrated based on spectral images captured underwater and in air. With the imaging model available, energy loss due to water attenuation is restored for images captured at different underwater distances. An experimental setup is built and experiments are carried out to verify the proposed method. Underwater images captured within an underwater distance of 260 cm are restored and compared with those in air. Results show that the relative restoration error is 3.58% on average for the test images, thus proving the accuracy of the proposed method. PMID:27410329

  9. Model-based restoration of underwater spectral images captured with narrowband filters.

    PubMed

    Guo, Yilu; Song, Hong; Liu, Hongbo; Wei, He; Yang, Ping; Zhan, Shuyue; Wang, Hangzhou; Huang, Hui; Liao, Ningfang; Mu, Quanquan; Leng, Jianxing; Yang, Wenjing

    2016-06-13

    Underwater spectral imaging is a promising method for mapping, classification and health monitoring of coral reefs and seafloor inhabitants. However, the spectrum of light is distorted during the underwater imaging process due to wavelength-dependent attenuation by the water. This paper presents a model-based method that accurately restores brightness of underwater spectral images captured with narrowband filters. A model is built for narrowband underwater spectral imaging. The model structure is derived from physical principles, representing the absorption, scattering and refraction by water and the optical properties of narrowband filters, lenses and image sensors. The model coefficients are calibrated based on spectral images captured underwater and in air. With the imaging model available, energy loss due to water attenuation is restored for images captured at different underwater distances. An experimental setup is built and experiments are carried out to verify the proposed method. Underwater images captured within an underwater distance of 260 cm are restored and compared with those in air. Results show that the relative restoration error is 3.58% on average for the test images, thus proving the accuracy of the proposed method.

  10. Spectral feature variations in x-ray diffraction imaging systems

    NASA Astrophysics Data System (ADS)

    Wolter, Scott D.; Greenberg, Joel A.

    2016-05-01

    Materials with different atomic or molecular structures give rise to unique scatter spectra when measured by X-ray diffraction. The details of these spectra, though, can vary based on both intrinsic (e.g., degree of crystallinity or doping) and extrinsic (e.g., pressure or temperature) conditions. While this sensitivity is useful for detailed characterizations of the material properties, these dependences make it difficult to perform more general classification tasks, such as explosives threat detection in aviation security. A number of challenges, therefore, currently exist for reliable substance detection including the similarity in spectral features among some categories of materials combined with spectral feature variations from materials processing and environmental factors. These factors complicate the creation of a material dictionary and the implementation of conventional classification and detection algorithms. Herein, we report on two prominent factors that lead to variations in spectral features: crystalline texture and temperature variations. Spectral feature comparisons between materials categories will be described for solid metallic sheet, aqueous liquids, polymer sheet, and metallic, organic, and inorganic powder specimens. While liquids are largely immune to texture effects, they are susceptible to temperature changes that can modify their density or produce phase changes. We will describe in situ temperature-dependent measurement of aqueous-based commercial goods in the temperature range of -20°C to 35°C.

  11. Stimulated Raman hyperspectral imaging based on spectral filtering of broadband fiber laser pulses.

    PubMed

    Ozeki, Yasuyuki; Umemura, Wataru; Sumimura, Kazuhiko; Nishizawa, Norihiko; Fukui, Kiichi; Itoh, Kazuyoshi

    2012-02-01

    We demonstrate a technique of hyperspectral imaging in stimulated Raman scattering (SRS) microscopy using a tunable optical filter, whose transmission wavelength can be varied quickly by a galvanometer mirror. Experimentally, broadband Yb fiber laser pulses are synchronized with picosecond Ti:sapphire pulses, and then spectrally filtered out by the filter. After amplification by fiber amplifiers, we obtain narrowband pulses with a spectral width of <3.3 cm(-1) and a wavelength tunability of >225 cm(-1). By using these pulses, we accomplish SRS imaging of polymer beads with spectral information.

  12. Stimulated Raman hyperspectral imaging based on spectral filtering of broadband fiber laser pulses.

    PubMed

    Ozeki, Yasuyuki; Umemura, Wataru; Sumimura, Kazuhiko; Nishizawa, Norihiko; Fukui, Kiichi; Itoh, Kazuyoshi

    2012-02-01

    We demonstrate a technique of hyperspectral imaging in stimulated Raman scattering (SRS) microscopy using a tunable optical filter, whose transmission wavelength can be varied quickly by a galvanometer mirror. Experimentally, broadband Yb fiber laser pulses are synchronized with picosecond Ti:sapphire pulses, and then spectrally filtered out by the filter. After amplification by fiber amplifiers, we obtain narrowband pulses with a spectral width of <3.3 cm(-1) and a wavelength tunability of >225 cm(-1). By using these pulses, we accomplish SRS imaging of polymer beads with spectral information. PMID:22297376

  13. Spectral compression algorithms for the analysis of very large multivariate images

    DOEpatents

    Keenan, Michael R.

    2007-10-16

    A method for spectrally compressing data sets enables the efficient analysis of very large multivariate images. The spectral compression algorithm uses a factored representation of the data that can be obtained from Principal Components Analysis or other factorization technique. Furthermore, a block algorithm can be used for performing common operations more efficiently. An image analysis can be performed on the factored representation of the data, using only the most significant factors. The spectral compression algorithm can be combined with a spatial compression algorithm to provide further computational efficiencies.

  14. Visible spectral imager for occultation and nightglow (VISION) for the PICASSO Mission

    NASA Astrophysics Data System (ADS)

    Saari, Heikki; Näsilä, Antti; Holmlund, Christer; Mannila, Rami; Näkki, Ismo; Ojanen, Harri J.; Fussen, Didier; Pieroux, Didier; Demoulin, Philippe; Dekemper, Emmanuel; Vanhellemont, Filip

    2015-10-01

    PICASSO - A PICo-satellite for Atmospheric and Space Science Observations is an ESA project led by the Belgian Institute for Space Aeronomy, in collaboration with VTT, Clyde Space Ltd. (UK), and the Centre Spatial de Liège (BE). VTT Technical Research Centre of Finland Ltd. will deliver the Visible Spectral Imager for Occultation and Nightglow (VISION) for the PICASSO mission. The VISION targets primarily the observation of the Earth's atmospheric limb during orbital Sun occultation. By assessing the radiation absorption in the Chappuis band for different tangent altitudes, the vertical profile of the ozone is retrieved. A secondary objective is to measure the deformation of the solar disk so that stratospheric and mesospheric temperature profiles are retrieved by inversion of the refractive raytracing problem. Finally, occasional full spectral observations of polar auroras are also foreseen. The VISION design realized with commercial of the shelf (CoTS) parts is described. The VISION instrument is small, lightweight (~500 g), Piezo-actuated Fabry-Perot Interferometer (PFPI) tunable spectral imager operating in the visible and near-infrared (430 - 800 nm). The spectral resolution over the whole wavelength range will be better than 10 nm @ FWHM. VISION has is 2.5° x 2.5° total field of view and it delivers maximum 2048 x 2048 pixel spectral images. The sun image size is around 0.5° i.e. ~500 pixels. To enable fast spectral data image acquisition VISION can be operated with programmable image sizes. VTT has previously developed PFPI tunable filter based AaSI Spectral Imager for the Aalto-1 Finnish CubeSat. In VISION the requirements of the spectral resolution and stability are tighter than in AaSI. Therefore the optimization of the of the PFPI gap control loop for the operating temperature range and vacuum conditions has to be improved. VISION optical, mechanical and electrical design is described.

  15. In-vivo full depth of eye imaging spectral domain optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Dai, Cuixia; Zhou, Chuanqing; Jiao, Shuliang; Xi, Peng; Ren, Qiushi

    2011-09-01

    It is necessary to apply the spectral-domain optical coherence tomography (SD-OCT) to image the whole eye segment for practically iatrical application, but the imaging depth of SD-OCT is limited by the spectral resolution of the spectrometer. By now, no result about this research has been reported. In our study, a new dual channel dual focus OCT system is adopted to image the whole eye segment. The cornea and the crystalline lens are simultaneously imaged by using full range complex spectral-domain OCT in one channel, the retina is detected by the other. The new system was successfully tested in imaging of the volunteer' eye in vivo. The preliminary results presented in this paper demonstrated the feasibility of this approach.

  16. High-speed imaging system based on spectral optical coherence tomography

    NASA Astrophysics Data System (ADS)

    Chen, Xiaodong; Li, Wanhui; Wang, Yi; Yu, Daoyin

    2005-01-01

    Optical coherence tomography (OCT) is an emerging technology for producing high-resolution cross-sectional imaging based on the principles of low-coherence interferometry. Tissue structure can be imaged on the micron scale in real time. The principle of OCT is analogous to that of ultrasound imaging, except that it measures the intensity of reflected infrared (IR) light rather than sound waves. A new spectral OCT(SOCT) technique is introduced. SOCT and its application to measure depth resolved spectral absorption is described. The crucial parameters of this method like transversal, depth, and spectral resolution and their relations are discussed. In combination with SOCT technique and endoscopes, SOCT enables high-resolution imaging in lumens of, for example, the esophagus and stomach. SOCT can be used to perform 'optical biopsy', or to guide surgery, by providing images in situ and in real time.

  17. Remote spectral imaging system (RSIS) based on an acousto-optic tunable filter (AOTF)

    SciTech Connect

    Moreau, F.; Hueber, D.M.; Vo-Dinh, T.

    1996-12-31

    This paper describes a new remote spectral imaging system (RSIS) based on an acousto-optic tunable filter (AOTF) capable of remote sensing using an imaging fiberoptic probe (IFP). A two-dimensional charge coupled device (CCD) was used as a detector. The AOTF was used as a wavelength selector. Unlike a tunable grating or prism based monochromator, the tunable filter has no moving parts, and it can be rapidly tuned to any wavelength in its operating range. The large aperture of the AOTF and its high spatial resolution allowed the optical image from an IFP to be recorded by a CCD. These characteristics, combined with their small size, make AOTF`s important new alternatives to conventional monochromators, especially for spectral multisensing and imaging. A prototype RSIS system, using both IFP and AOTF, was developed and its feasibility for spectral imaging was demonstrated. 40 refs., 8 figs.

  18. Use of Sonicated Albumin (Infoson) to Enhance Arterial Spectral and Color Doppler Imaging

    SciTech Connect

    Abildgaard, Andreas; Egge, Tor S.; Klow, Nils-Einar; Jakobsen, Jarl A.

    1996-04-15

    Purpose: To examine the effect of an ultrasound contrast medium (UCM), Infoson, on Doppler examination of stenotic arteries. Methods: Stenoses were created in the common carotid artery of six piglets, and examined with spectral Doppler and color Doppler imaging during UCM infusion in the left ventricle. Results: UCM caused a mean increase in recorded maximal systolic and end-diastolic velocities of 5% and 6%, respectively, while blood flow remained constant. Increased spectral intensity with UCM was accompanied by spectral broadening. Reduction of spectral intensity by adjustment of Doppler gain counteracted the velocity effects and the spectral broadening. With color Doppler, UCM caused dose-dependent color artifacts outside the artery. Flow in narrow stenoses could be visualized with UCM. Conclusion: The effects of UCM on velocity measurements were slight, and were related to changes in spectral intensity. With color Doppler, UCM may facilitate flow detection, but color artifacts may interfere.

  19. Pigment Identification of Paintings Based on Kubelka-Munk Theory and Spectral Images

    NASA Astrophysics Data System (ADS)

    Moghareh Abed, Farhad

    The preservation of cultural heritage and treatment thereof are delicate responsibilities that demand the best possible technologies and extreme care to avoid any irreversible loss. It necessitates a deep understanding of constituent materials, along with the analytical methods and cutting-edge technologies. Considering this direction, the goal of this dissertation is to promote the conservation procedures by providing an applicable workflow for spectral-based pigment identification. The proposed pipeline is a novel and practical aid for museum conservators for many aspects, such as inpainting, treatment and archiving of artwork. Spectral-based pigment identification algorithms rely on accurate spectral data, a subtractive mixing model and an effective unmixing algorithm. In this dissertation, the spectral data were obtained using a spectral image acquisition system as a feasible and non-destructive technique. A liquid-crystal tunable filter (LCTF) and a CCD camera were used for spectral measurement of the painting. The spectral accuracy and precision of the LCTF-based spectral acquisition system were assessed and enhanced. Of the common factors affecting the acquisition performance, capturing geometry, LCTF angular dependencies and spectral characterization algorithm were new contributions to the traditional workflow. The complexity of subtractive mixtures limits the effective application of linear unmixing algorithms for pigment identification. Accordingly, a new linear modification of single-constant Kubelka-Munk theory was derived to enable the use of available linear spectral unmixing algorithms for paint mixtures. A selection of geometric and iterative-based unmixing algorithms was applied to the LCTF spectral images in the subtractive mixing space using the defined subtractive linear model. Final sets of primary pigments were improved employing a pre-existing database of common pigments as a tool for practical inpainting procedures. The pigment maps, showing

  20. Multiphoton microscopy of cleared mouse organs

    NASA Astrophysics Data System (ADS)

    Parra, Sonia G.; Chia, Thomas H.; Zinter, Joseph P.; Levene, Michael J.

    2010-05-01

    Typical imaging depths with multiphoton microscopy (MPM) are limited to less than 300 μm in many tissues due to light scattering. Optical clearing significantly reduces light scattering by replacing water in the organ tissue with a fluid having a similar index of refraction to that of proteins. We demonstrate MPM of intact, fixed, cleared mouse organs with penetration depths and fields of view in excess of 2 mm. MPM enables the creation of large 3-D data sets with flexibility in pixel format and ready access to intrinsic fluorescence and second-harmonic generation. We present high-resolution images and 3-D image stacks of the brain, small intestine, large intestine, kidney, lung, and testicle with image sizes as large as 4096×4096 pixels.

  1. Point spread function engineering with multiphoton SPIFI

    NASA Astrophysics Data System (ADS)

    Wernsing, Keith A.; Field, Jeffrey J.; Domingue, Scott R.; Allende-Motz, Alyssa M.; DeLuca, Keith F.; Levi, Dean H.; DeLuca, Jennifer G.; Young, Michael D.; Squier, Jeff A.; Bartels, Randy A.

    2016-03-01

    MultiPhoton SPatIal Frequency modulated Imaging (MP-SPIFI) has recently demonstrated the ability to simultaneously obtain super-resolved images in both coherent and incoherent scattering processes -- namely, second harmonic generation and two-photon fluorescence, respectively.1 In our previous analysis, we considered image formation produced by the zero and first diffracted orders from the SPIFI modulator. However, the modulator is a binary amplitude mask, and therefore produces multiple diffracted orders. In this work, we extend our analysis to image formation in the presence of higher diffracted orders. We find that tuning the mask duty cycle offers a measure of control over the shape of super-resolved point spread functions in an MP-SPIFI microscope.

  2. Methodology for diagnosing of skin cancer on images of dermatologic spots by spectral analysis

    PubMed Central

    Guerra-Rosas, Esperanza; Álvarez-Borrego, Josué

    2015-01-01

    In this paper a new methodology for the diagnosing of skin cancer on images of dermatologic spots using image processing is presented. Currently skin cancer is one of the most frequent diseases in humans. This methodology is based on Fourier spectral analysis by using filters such as the classic, inverse and k-law nonlinear. The sample images were obtained by a medical specialist and a new spectral technique is developed to obtain a quantitative measurement of the complex pattern found in cancerous skin spots. Finally a spectral index is calculated to obtain a range of spectral indices defined for skin cancer. Our results show a confidence level of 95.4%. PMID:26504638

  3. Principal Components Analysis of Martian NIR Image Cubes to Retrieve Surface Spectral Endmembers

    NASA Astrophysics Data System (ADS)

    Klassen, David R.

    2016-07-01

    Presented here is a discussion of the complete principal components analysis (PCA) performed on all photometric NASA Infrared Telescope Facility (IRTF) NSFCAM spectral image sets from 1995–2001 and Mars Reconnaissance Orbiter (MRO) Compact Reconnaissance Imaging Spectrometer for Mars (CRISM) spectral image sets from 2006–2008, detailing the similarities and differences and overall interpretation of the PC dimensional spaces. The purpose of the analysis is to use the PCA to recover surface spectral endmembers to be used in a full radiative transfer modeling program to recover ice cloud optical depths (and thus water content) over diurnal, seasonal, and interannual timescales. The PCA results show considerable consistency across all seasons, and can be optimized to increase the consistency through both spectral and geographic restrictions on the data.

  4. Principal Components Analysis of Martian NIR Image Cubes to Retrieve Surface Spectral Endmembers

    NASA Astrophysics Data System (ADS)

    Klassen, David R.

    2016-07-01

    Presented here is a discussion of the complete principal components analysis (PCA) performed on all photometric NASA Infrared Telescope Facility (IRTF) NSFCAM spectral image sets from 1995-2001 and Mars Reconnaissance Orbiter (MRO) Compact Reconnaissance Imaging Spectrometer for Mars (CRISM) spectral image sets from 2006-2008, detailing the similarities and differences and overall interpretation of the PC dimensional spaces. The purpose of the analysis is to use the PCA to recover surface spectral endmembers to be used in a full radiative transfer modeling program to recover ice cloud optical depths (and thus water content) over diurnal, seasonal, and interannual timescales. The PCA results show considerable consistency across all seasons, and can be optimized to increase the consistency through both spectral and geographic restrictions on the data.

  5. Biomedical Applications of the Information-efficient Spectral Imaging Sensor (ISIS)

    SciTech Connect

    Gentry, S.M.; Levenson, R.

    1999-01-21

    The Information-efficient Spectral Imaging Sensor (ISIS) approach to spectral imaging seeks to bridge the gap between tuned multispectral and fixed hyperspectral imaging sensors. By allowing the definition of completely general spectral filter functions, truly optimal measurements can be made for a given task. These optimal measurements significantly improve signal-to-noise ratio (SNR) and speed, minimize data volume and data rate, while preserving classification accuracy. The following paper investigates the application of the ISIS sensing approach in two sample biomedical applications: prostate and colon cancer screening. It is shown that in these applications, two to three optimal measurements are sufficient to capture the majority of classification information for critical sample constituents. In the prostate cancer example, the optimal measurements allow 8% relative improvement in classification accuracy of critical cell constituents over a red, green, blue (RGB) sensor. In the colon cancer example, use of optimal measurements boost the classification accuracy of critical cell constituents by 28% relative to the RGB sensor. In both cases, optimal measurements match the performance achieved by the entire hyperspectral data set. The paper concludes that an ISIS style spectral imager can acquire these optimal spectral images directly, allowing improved classification accuracy over an RGB sensor. Compared to a hyperspectral sensor, the ISIS approach can achieve similar classification accuracy using a significantly lower number of spectral samples, thus minimizing overall sample classification time and cost.

  6. The Spectral Image Processing System (SIPS): Software for integrated analysis of AVIRIS data

    NASA Technical Reports Server (NTRS)

    Kruse, F. A.; Lefkoff, A. B.; Boardman, J. W.; Heidebrecht, K. B.; Shapiro, A. T.; Barloon, P. J.; Goetz, A. F. H.

    1992-01-01

    The Spectral Image Processing System (SIPS) is a software package developed by the Center for the Study of Earth from Space (CSES) at the University of Colorado, Boulder, in response to a perceived need to provide integrated tools for analysis of imaging spectrometer data both spectrally and spatially. SIPS was specifically designed to deal with data from the Airborne Visible/Infrared Imaging Spectrometer (AVIRIS) and the High Resolution Imaging Spectrometer (HIRIS), but was tested with other datasets including the Geophysical and Environmental Research Imaging Spectrometer (GERIS), GEOSCAN images, and Landsat TM. SIPS was developed using the 'Interactive Data Language' (IDL). It takes advantage of high speed disk access and fast processors running under the UNIX operating system to provide rapid analysis of entire imaging spectrometer datasets. SIPS allows analysis of single or multiple imaging spectrometer data segments at full spatial and spectral resolution. It also allows visualization and interactive analysis of image cubes derived from quantitative analysis procedures such as absorption band characterization and spectral unmixing. SIPS consists of three modules: SIPS Utilities, SIPS_View, and SIPS Analysis. SIPS version 1.1 is described below.

  7. Imaging tamoxifen retinopathy using spectral-domain optical coherence tomography

    PubMed Central

    Caramoy, Albert; Scholz, Paula; Fauser, Sascha; Kirchhof, Bernd

    2011-01-01

    A case of tamoxifen retinopathy examined with spectral-domain optical coherence tomography (SD-OCT) is presented. The typical refractile deposits are located between ganglion cell layer and inner plexiform layer in SD-OCT. A defect on the outer retinal layer with disruption of the photoreceptor layer with sharp edges is seen. The still attached posterior hyaloids gives evidence of other pathomechanism involved in the outer retinal defect than that of macular hole, as suggested in the literature.

  8. Present and future status of flexible spectral imaging color enhancement and blue laser imaging technology.

    PubMed

    Osawa, Hiroyuki; Yamamoto, Hironori

    2014-01-01

    The usefulness of flexible spectral imaging color enhancement (FICE) has been reported for evaluating the esophagus, stomach, and small and large intestine. Higher contrast is shown between cancer and the surrounding mucosa in the esophagus and stomach and may facilitate the detection of gastric cancers missed by white light imaging alone. The surface patterns of gastric mucosa are clearly visualized in non-malignant areas but are irregular and blurred in malignant areas, leading to clear demarcation. Capsule endoscopy with FICE detects angiodysplasia and erosions of the small intestine. The surface and vascular pattern with FICE is useful for the differential diagnosis of colorectal polyps. However, FICE remains somewhat poor at visualizing mucosal microvasculature on a tumor surface. Narrow-band imaging (NBI) is dark in observing whole gastric mucosa and poor at visualizing mucosal microstructure. Blue laser imaging (BLI) has the potential to resolve these limitations. Narrow-band laser light combined with white light shows irregular microvessels on both differentiated and undifferentiated gastric cancer similar to those using NBI. In addition, irregular surface patterns including minute white zones are clearly seen on the uneven surface of differentiated lesions, resulting in exclusion of undifferentiated lesions. Using both distant and close-up views, a high contrast between green intestinal metaplasia and brown gastric cancer may lead to early detection of gastric cancers and determination of a demarcation line. BLI produces high-contrast images in esophageal cancer with clear vision of intrapapillary capillary loops and also predicts the histopathological diagnosis and depth of invasion in colorectal neoplasms.

  9. MARS spectral molecular imaging of lamb tissue: data collection and image analysis

    NASA Astrophysics Data System (ADS)

    Aamir, R.; Chernoglazov, A.; Bateman, C. J.; Butler, A. P. H.; Butler, P. H.; Anderson, N. G.; Bell, S. T.; Panta, R. K.; Healy, J. L.; Mohr, J. L.; Rajendran, K.; Walsh, M. F.; de Ruiter, N.; Gieseg, S. P.; Woodfield, T.; Renaud, P. F.; Brooke, L.; Abdul-Majid, S.; Clyne, M.; Glendenning, R.; Bones, P. J.; Billinghurst, M.; Bartneck, C.; Mandalika, H.; Grasset, R.; Schleich, N.; Scott, N.; Nik, S. J.; Opie, A.; Janmale, T.; Tang, D. N.; Kim, D.; Doesburg, R. M.; Zainon, R.; Ronaldson, J. P.; Cook, N. J.; Smithies, D. J.; Hodge, K.

    2014-02-01

    Spectral molecular imaging is a new imaging technique able to discriminate and quantify different components of tissue simultaneously at high spatial and high energy resolution. Our MARS scanner is an x-ray based small animal CT system designed to be used in the diagnostic energy range (20-140 keV). In this paper, we demonstrate the use of the MARS scanner, equipped with the Medipix3RX spectroscopic photon-processing detector, to discriminate fat, calcium, and water in tissue. We present data collected from a sample of lamb meat including bone as an illustrative example of human tissue imaging. The data is analyzed using our 3D Algebraic Reconstruction Algorithm (MARS-ART) and by material decomposition based on a constrained linear least squares algorithm. The results presented here clearly show the quantification of lipid-like, water-like and bone-like components of tissue. However, it is also clear to us that better algorithms could extract more information of clinical interest from our data. Because we are one of the first to present data from multi-energy photon-processing small animal CT systems, we make the raw, partial and fully processed data available with the intention that others can analyze it using their familiar routines. The raw, partially processed and fully processed data of lamb tissue along with the phantom calibration data can be found at http://hdl.handle.net/10092/8531.

  10. Multi-spectral remote sensing image retrieval based on semantic extraction

    NASA Astrophysics Data System (ADS)

    Liu, Tingting; Li, Pingxiang; Zhang, Liangpei; Chen, Xu

    2008-12-01

    In this paper, a multi-level image representation model is developed and used for multi-spectral remote sensing image retrieval in order to narrow the gap between the low-level feature and high-level semantic. This model consists of an image segmentation part, a feature extraction part and semantic extraction part. The first two parts aim at the extraction of primitive region feature of an image. In these two steps, an improved JSEG algorithm is used to segment the image stored in the database, then spectral feature and texture feature are extracted for each region. In semantic extraction part, the semantic information hidden in different regions of different images is extracted by Bayesian method and expectation maximization (EM) method. At last, positive example and negative example concept is used in image retrieval instead of relevant feedback. Experiment shows that this method not only improves the accuracy of the result but also decreases the complexity of retrieval.

  11. Multi-spectral imaging with infrared sensitive organic light emitting diode.

    PubMed

    Kim, Do Young; Lai, Tzung-Han; Lee, Jae Woong; Manders, Jesse R; So, Franky

    2014-01-01

    Commercially available near-infrared (IR) imagers are fabricated by integrating expensive epitaxial grown III-V compound semiconductor sensors with Si-based readout integrated circuits (ROIC) by indium bump bonding which significantly increases the fabrication costs of these image sensors. Furthermore, these typical III-V compound semiconductors are not sensitive to the visible region and thus cannot be used for multi-spectral (visible to near-IR) sensing. Here, a low cost infrared (IR) imaging camera is demonstrated with a commercially available digital single-lens reflex (DSLR) camera and an IR sensitive organic light emitting diode (IR-OLED). With an IR-OLED, IR images at a wavelength of 1.2 µm are directly converted to visible images which are then recorded in a Si-CMOS DSLR camera. This multi-spectral imaging system is capable of capturing images at wavelengths in the near-infrared as well as visible regions. PMID:25091589

  12. Multi-spectral imaging with infrared sensitive organic light emitting diode

    NASA Astrophysics Data System (ADS)

    Kim, Do Young; Lai, Tzung-Han; Lee, Jae Woong; Manders, Jesse R.; So, Franky

    2014-08-01

    Commercially available near-infrared (IR) imagers are fabricated by integrating expensive epitaxial grown III-V compound semiconductor sensors with Si-based readout integrated circuits (ROIC) by indium bump bonding which significantly increases the fabrication costs of these image sensors. Furthermore, these typical III-V compound semiconductors are not sensitive to the visible region and thus cannot be used for multi-spectral (visible to near-IR) sensing. Here, a low cost infrared (IR) imaging camera is demonstrated with a commercially available digital single-lens reflex (DSLR) camera and an IR sensitive organic light emitting diode (IR-OLED). With an IR-OLED, IR images at a wavelength of 1.2 µm are directly converted to visible images which are then recorded in a Si-CMOS DSLR camera. This multi-spectral imaging system is capable of capturing images at wavelengths in the near-infrared as well as visible regions.

  13. HIGH THROUGHPUT SPECTRAL IMAGING SYSTEM FOR WHOLESOMENESS INSPECTION OF CHICKEN

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An online line-scan imaging system containing an electron-multiplying charge-coupled device detector and line-scan spectrograph was used for identifying wholesome and unwholesome freshly slaughtered chicken carcasses on high-speed commercial chicken processing lines. Hyperspectral images were acqui...

  14. Inference of dense spectral reflectance images from sparse reflectance measurement using non-linear regression modeling

    NASA Astrophysics Data System (ADS)

    Deglint, Jason; Kazemzadeh, Farnoud; Wong, Alexander; Clausi, David A.

    2015-09-01

    One method to acquire multispectral images is to sequentially capture a series of images where each image contains information from a different bandwidth of light. Another method is to use a series of beamsplitters and dichroic filters to guide different bandwidths of light onto different cameras. However, these methods are very time consuming and expensive and perform poorly in dynamic scenes or when observing transient phenomena. An alternative strategy to capturing multispectral data is to infer this data using sparse spectral reflectance measurements captured using an imaging device with overlapping bandpass filters, such as a consumer digital camera using a Bayer filter pattern. Currently the only method of inferring dense reflectance spectra is the Wiener adaptive filter, which makes Gaussian assumptions about the data. However, these assumptions may not always hold true for all data. We propose a new technique to infer dense reflectance spectra from sparse spectral measurements through the use of a non-linear regression model. The non-linear regression model used in this technique is the random forest model, which is an ensemble of decision trees and trained via the spectral characterization of the optical imaging system and spectral data pair generation. This model is then evaluated by spectrally characterizing different patches on the Macbeth color chart, as well as by reconstructing inferred multispectral images. Results show that the proposed technique can produce inferred dense reflectance spectra that correlate well with the true dense reflectance spectra, which illustrates the merits of the technique.

  15. Enhancement of infrared spectral images for maximizing chemical information by minimizing baseline interferences.

    PubMed

    Bu, Dongsheng; Huffman, Scott W; Seelenbinder, John A; Brown, Chris W

    2005-05-01

    The popularity of spectral images in many areas of analysis has greatly increased during the last decade due to the development of charge-coupled device (CCD) and infrared sensitive cameras. Large amounts of spatial information can be obtained in short periods of time. The general goal in analytical chemistry is to convert spectral images into chemical images, which show the spatial locations of various chemical components. Self-modeling multivariate curve resolution methods can be used to extract pure component spectra from the mixture spectra in images and produce chemical images. However, there is a difficulty in processing infrared spectral images due to large pixel-to-pixel baseline variations. Herein, a method for minimizing baseline interferences using fast Fourier transform (FFT) filtering in both the spectral and spatial domains is discussed. The methodology is demonstrated on a microscopic sample of butter contaminated with non-pathogenic E. coli and on a cross-sectional sample of rabbit aorta containing plaque. The processing to reduce baseline effects improved the spatial resolution without compromising the spectral resolution.

  16. Principles and satellite applications of the information-efficient spectral imaging sensor

    SciTech Connect

    Stallard, B.R.; Gentry, S.M.; Sweatt, W.C.; Motomatsu, S.E.; Boye, C.A.

    1997-06-01

    Remote sensing by satellite is increasingly important to the national government for treaty verification, battlefield monitoring, and other activities. In addition, civilian oriented applications are increasing in areas such as geology, meteorology, ecology, forestry, and agriculture. Spectral imaging sensors, an important subclass of satellite-borne sensors, have been shown to provide information far superior to that of conventional panchromatic images in many of these applications. However, spectral imaging adds at least two challenges to the already difficult task of viewing the earth from a distance of hundreds of kilometers. First,