Detecting Bots on Russian Political Twitter.
Stukal, Denis; Sanovich, Sergey; Bonneau, Richard; Tucker, Joshua A
2017-12-01
Automated and semiautomated Twitter accounts, bots, have recently gained significant public attention due to their potential interference in the political realm. In this study, we develop a methodology for detecting bots on Twitter using an ensemble of classifiers and apply it to study bot activity within political discussions in the Russian Twittersphere. We focus on the interval from February 2014 to December 2015, an especially consequential period in Russian politics. Among accounts actively Tweeting about Russian politics, we find that on the majority of days, the proportion of Tweets produced by bots exceeds 50%. We reveal bot characteristics that distinguish them from humans in this corpus, and find that the software platform used for Tweeting is among the best predictors of bots. Finally, we find suggestive evidence that one prominent activity that bots were involved in on Russian political Twitter is the spread of news stories and promotion of media who produce them.
Multimodal game bot detection using user behavioral characteristics.
Kang, Ah Reum; Jeong, Seong Hoon; Mohaisen, Aziz; Kim, Huy Kang
2016-01-01
As the online service industry has continued to grow, illegal activities in the online world have drastically increased and become more diverse. Most illegal activities occur continuously because cyber assets, such as game items and cyber money in online games, can be monetized into real currency. The aim of this study is to detect game bots in a massively multiplayer online role playing game (MMORPG). We observed the behavioral characteristics of game bots and found that they execute repetitive tasks associated with gold farming and real money trading. We propose a game bot detection method based on user behavioral characteristics. The method of this paper was applied to real data provided by a major MMORPG company. Detection accuracy rate increased to 96.06 % on the banned account list.
AntBot: Anti-pollution peer-to-peer botnets
DOE Office of Scientific and Technical Information (OSTI.GOV)
Yan, Guanhua; Eidenbenz, Stephan; Ha, Duc T
2009-01-01
Botnets, which are responsible for many email sparnming and DDoS (Distributed Denial of Service) attacks in the current Internet, have emerged as one of most severe cyber-threats in recent years. To evade detection and improve resistance against countermeasures, botnets have evolved from the first generation that relies on IRC chat channels to deliver commands to the current generation that uses highly resilient P2P (Peer-to-Peer) protocols to spread their C&C (Command and Control) information. It is, however, revealed that P2P botnets, although relieved from the single point of failure that IRC botnets suffer, can be easily disrupted using pollution-based mitigation schemesmore » [15]. In this paper, we play the devil's advocate and propose a new type of hypothetical botnets called AntBot, which aim to propagate their C&C information to individual bots even though there exists an adversary that persistently pollutes keys used by seized bots to search the command information. The key idea of AntBot is a tree-like structure that bots use to deliver the command so that captured bots reveal only limited information. To evaluate effectiveness of AntBot against pollution-based mitigation in a virtual environment, we develop a distributed P2P botnet simulator. Using extensive experiments, we demonstrate that AntBot operates resiliently against pollution-based mitigation. We further present a few potential defense schemes that could effectively disrupt AntBot operations.« less
Social Bots: Human-Like by Means of Human Control?
Grimme, Christian; Preuss, Mike; Adam, Lena; Trautmann, Heike
2017-12-01
Social bots are currently regarded an influential but also somewhat mysterious factor in public discourse and opinion making. They are considered to be capable of massively distributing propaganda in social and online media, and their application is even suspected to be partly responsible for recent election results. Astonishingly, the term social bot is not well defined and different scientific disciplines use divergent definitions. This work starts with a balanced definition attempt, before providing an overview of how social bots actually work (taking the example of Twitter) and what their current technical limitations are. Despite recent research progress in Deep Learning and Big Data, there are many activities bots cannot handle well. We then discuss how bot capabilities can be extended and controlled by integrating humans into the process and reason that this is currently the most promising way to realize meaningful interactions with other humans. This finally leads to the conclusion that hybridization is a challenge for current detection mechanisms and has to be handled with more sophisticated approaches to identify political propaganda distributed with social bots.
Even good bots fight: The case of Wikipedia.
Tsvetkova, Milena; García-Gavilanes, Ruth; Floridi, Luciano; Yasseri, Taha
2017-01-01
In recent years, there has been a huge increase in the number of bots online, varying from Web crawlers for search engines, to chatbots for online customer service, spambots on social media, and content-editing bots in online collaboration communities. The online world has turned into an ecosystem of bots. However, our knowledge of how these automated agents are interacting with each other is rather poor. Bots are predictable automatons that do not have the capacity for emotions, meaning-making, creativity, and sociality and it is hence natural to expect interactions between bots to be relatively predictable and uneventful. In this article, we analyze the interactions between bots that edit articles on Wikipedia. We track the extent to which bots undid each other's edits over the period 2001-2010, model how pairs of bots interact over time, and identify different types of interaction trajectories. We find that, although Wikipedia bots are intended to support the encyclopedia, they often undo each other's edits and these sterile "fights" may sometimes continue for years. Unlike humans on Wikipedia, bots' interactions tend to occur over longer periods of time and to be more reciprocated. Yet, just like humans, bots in different cultural environments may behave differently. Our research suggests that even relatively "dumb" bots may give rise to complex interactions, and this carries important implications for Artificial Intelligence research. Understanding what affects bot-bot interactions is crucial for managing social media well, providing adequate cyber-security, and designing well functioning autonomous vehicles.
Even good bots fight: The case of Wikipedia
Tsvetkova, Milena; García-Gavilanes, Ruth; Floridi, Luciano; Yasseri, Taha
2017-01-01
In recent years, there has been a huge increase in the number of bots online, varying from Web crawlers for search engines, to chatbots for online customer service, spambots on social media, and content-editing bots in online collaboration communities. The online world has turned into an ecosystem of bots. However, our knowledge of how these automated agents are interacting with each other is rather poor. Bots are predictable automatons that do not have the capacity for emotions, meaning-making, creativity, and sociality and it is hence natural to expect interactions between bots to be relatively predictable and uneventful. In this article, we analyze the interactions between bots that edit articles on Wikipedia. We track the extent to which bots undid each other’s edits over the period 2001–2010, model how pairs of bots interact over time, and identify different types of interaction trajectories. We find that, although Wikipedia bots are intended to support the encyclopedia, they often undo each other’s edits and these sterile “fights” may sometimes continue for years. Unlike humans on Wikipedia, bots’ interactions tend to occur over longer periods of time and to be more reciprocated. Yet, just like humans, bots in different cultural environments may behave differently. Our research suggests that even relatively “dumb” bots may give rise to complex interactions, and this carries important implications for Artificial Intelligence research. Understanding what affects bot-bot interactions is crucial for managing social media well, providing adequate cyber-security, and designing well functioning autonomous vehicles. PMID:28231323
Identifying MMORPG Bots: A Traffic Analysis Approach
NASA Astrophysics Data System (ADS)
Chen, Kuan-Ta; Jiang, Jhih-Wei; Huang, Polly; Chu, Hao-Hua; Lei, Chin-Laung; Chen, Wen-Chin
2008-12-01
Massively multiplayer online role playing games (MMORPGs) have become extremely popular among network gamers. Despite their success, one of MMORPG's greatest challenges is the increasing use of game bots, that is, autoplaying game clients. The use of game bots is considered unsportsmanlike and is therefore forbidden. To keep games in order, game police, played by actual human players, often patrol game zones and question suspicious players. This practice, however, is labor-intensive and ineffective. To address this problem, we analyze the traffic generated by human players versus game bots and propose general solutions to identify game bots. Taking Ragnarok Online as our subject, we study the traffic generated by human players and game bots. We find that their traffic is distinguishable by 1) the regularity in the release time of client commands, 2) the trend and magnitude of traffic burstiness in multiple time scales, and 3) the sensitivity to different network conditions. Based on these findings, we propose four strategies and two ensemble schemes to identify bots. Finally, we discuss the robustness of the proposed methods against countermeasures of bot developers, and consider a number of possible ways to manage the increasingly serious bot problem.
RatBot: anti-enumeration peer-to-peer botnets
DOE Office of Scientific and Technical Information (OSTI.GOV)
Yan, Guanhua; Eidenbenz, Stephan; Chen, Songqing
2010-01-01
Botnets have emerged as one of the most severe cyber threats in recent years. To obtain high resilience against a single point of failure, the new generation of botnets have adopted the peer-to-peer (P2P) structure. One critical question regarding these P2P botnets is: how big are they indeed? To address this question, researchers have proposed both actively crawling and passively monitoring methods to enumerate existing P2P botnets. In this work, we go further to explore the potential strategies that botnets may have to obfuscate their true sizes. Towards this end, this paper introduces RatBot, a P2P botnet that applies somemore » statistical techniques to defeat existing P2P botnet enumeration methods. The key ideas of RatBot are two-fold: (1) there exist a fraction of bots that are indistinguishable from their fake identities, which are spoofing IP addresses they use to hide themselves; (2) we use a heavy-tailed distribution to generate the number of fake identities for each of these bots so that the sum of observed fake identities converges only slowly and thus has high variation. We use large-scale high-fidelity simulation to quantify the estimation errors under diverse settings, and the results show that a naive enumeration technique can overestimate the sizes of P2P botnets by one order of magnitude. We believe that our work reveals new challenges of accurately estimating the sizes of P2P botnets, and hope that it will raise the awareness of security practitioners with these challenges. We further suggest a few countermeasures that can potentially defeat RatBot's anti-enumeration scheme.« less
Horn, Lars-Christian; Angermann, Karolin; Hentschel, Bettina; Einenkel, Jens; Höhn, Anne Kathrin
2017-04-01
Borderline ovarian tumors (BOT) arise from cystadenomas and represent a transition step within the development of low-grade ovarian carcinomas (Type I tumors). That pathway mirrors the adenoma-to-carcinoma sequence known for colorectal cancer. It has been suggested that papillary tubal hyperplasia (PTH) and salpingoliths may be associated with the development of BOT. To evaluate the frequency of the presence of benign cystadenoma and its transition to BOT in a given patient as well as the presence of PTH and salpingoliths we re-valuated in 74 consecutive cases of BOT with different histologic types. The majority of cases represented serous-BOT (60.8%), followed by mucinous BOT (25.7%), other histologic types were rare. 86.5% showed an adenoma-BOT sequence, which was seen in all mucinous BOT but was missed in 15.6% of serous BOT. Two cases had salpingoliths without associated PTH. PTH was seen in four out of the 74 (5.4%) BOT and occurred only in cases with serous histology. The vast majority of BOT represent a transition from benign cystadenoma to BOT in cases with mucinous and serous histology. Salpingoliths are rarely seen in association with BOT and occurred exclusively in BOT with serous histology. PTH may represent a distinct lesion but is rarely seen in association with BOT, especially in those with non-serous histology. Further studies are needed to evaluate the frequency and pathogenetic association of PTH with BOT. Copyright © 2017 Elsevier GmbH. All rights reserved.
Product Searching with Shopping Bots.
ERIC Educational Resources Information Center
Rowley, Jennifer
2000-01-01
Using trial searches for three best-selling books, this study examined the search facilities offered by shopping bots, which support consumers with the product search and identification stage in e-shopping. Findings indicate that effectiveness of bots not only depends upon search facilities but also upon product coverage, and other added value…
Tactical assessment in a squad of intelligent bots
NASA Astrophysics Data System (ADS)
Gołuński, Marcel; Wasiewicz, Piotr
2010-09-01
In this paper we explore the problem of communication and coordination in a team of intelligent game bots (aka embodied agents). It presents a tactical decision making system controlling the behavior of an autonomous bot followed by the concept of a team tactical decision making system controlling the team of intelligent bots. The algorithms to be introduced have been implemented in the Java language by means of Pogamut 2 framework, interfacing the bot logic with Unreal Tournament 2004 virtual environment.
TickBot: a novel robotic device for controlling tick populations in the natural environment.
Gaff, Holly D; White, Alexis; Leas, Kyle; Kelman, Pamela; Squire, James C; Livingston, David L; Sullivan, Gerald A; Baker, Elizabeth W; Sonenshine, Daniel E
2015-03-01
A semi-autonomous 4-wheeled robot (TickBot) was fitted with a denim cloth treated with an acaricide (permethrin™) and tested for its ability to control ticks in a tick-infested natural environment in Portsmouth, Virginia. The robot's sensors detect a magnetic field signal from a guide wire encased in 80m polyethylene tubing, enabling the robot to follow the trails, open areas and other terrain where the tubing was located. To attract ticks to the treated area, CO2 was distributed through the same tubing, fitted with evenly spaced pores and flow control valves, which permitted uniform CO2 distribution. Tests were done to determine the optimum frequency for TickBot to traverse the wire-guided treatment site as well as the duration of operation that could be accomplished on a single battery charge. Prior to treatment, dragging was done to determine the natural abundance of ticks in the test site. Controls were done without CO2 and without permethrin. TickBot proved highly effective in reducing the overall tick densities to nearly zero with the treatment that included both carbon dioxide pretreatment and the permethrin treated cloth. Following a 60min traverse of the treatment areas, adult tick numbers, almost entirely Amblyomma americanum, was reduced to zero within 1h and remained at or near zero for 24h. Treatments without CO2 also showed reduction of ticks to near zero within 1h, but the populations were no different than the control sections at 4h. This study demonstrates the efficacy of TickBot as a tick control device to significantly reduce the risk of tick bites and disease transmission to humans and companion animals visiting a previously tick-infested natural environment. Continued deployment of TickBot for additional days or weeks can assure a relatively tick-safe environment for enjoyment by the public. Copyright © 2014 Elsevier GmbH. All rights reserved.
Design and Development of Mopping Robot-'HotBot'
NASA Astrophysics Data System (ADS)
Khan, M. R.; Huq, N. M. L.; Billah, M. M.; Ahmmad, S. M.
2013-12-01
To have a healthy, comfortable, and fresh civilized life we need to do some unhealthy households. Cleaning the dirty floor with a mop is one of the most disgusting and scary house hold jobs. Mopping robots are a solution of such problem. However, these robots are still on the way to be smart enough. Many points limit their efficiency, i.e. cleaning sticky dirt, having dry floor after cleaning, monitoring, cost etc. 'HotBot' is a mopping robot that can clean dirty floor with nice efficiency leaving no sticky dirt. Hot water can be used for heavy stains or normal water for usual situation and economy. It needs neither to be monitored during mopping nor to wipe the floor after it. 'HotBot' has sensors to detect obstacles and a control mechanism to avoid those. Moreover, it cleans sequentially and equipped with several accident-protection-systems. Moreover, it is also cost effective compared to the robots available so far.
Karlsen, Nikoline Marie Schou; Karlsen, Mona Aarenstrup; Høgdall, Estrid; Nedergaard, Lotte; Christensen, Ib Jarle; Høgdall, Claus
2016-07-01
The aim of the study was to evaluate the rate of relapse as well as disease-free, overall, and disease-specific survival in women with borderline ovarian tumour (BOT). Furthermore, the study aims to identify the clinical parameters correlated to relapse. National clinical data of women diagnosed with BOT from January 2005 to January 2013 constituted the basis for our study population. The prognostic influence of clinical variables was evaluated using univariate and multivariate analyses. A total of 1143 women were eligible for analysis, with 87.9% in FIGO stage I and 12.1% in FIGO stages II-IV. Relapse of BOT was detected in 3.7%, hereof 40.5% with malignant transformation. The five-year disease-free survival was 97.6% in FIGO stage I and 87.3% in FIGO stages II-IV. Younger age, laparoscopic surgical approach, fertility sparing surgery, FIGO stages II-IV, bilateral tumour presence, serous histology, implants and microinvasion of the tumour were significantly associated with relapse in univariate analyses. The overall five-year survival rate was 92.2% in FIGO stage I and 89.0% in FIGO stages II-IV. Out of 77 deaths in total, only seven women died from BOT. A general favourable prognosis in women with BOT was confirmed in our study. Our findings indicate that systematic, long-term follow-up does not seem necessary in women treated for FIGO stage IA BOT with no residual disease or microinvasion. Copyright © 2016 Elsevier Inc. All rights reserved.
Energetic cost of bot fly parasitism in free-ranging eastern chipmunks.
Careau, Vincent; Thomas, Donald W; Humphries, Murray M
2010-02-01
The energy and nutrient demands of parasites on their hosts are frequently invoked as an explanation for negative impacts of parasitism on host survival and reproductive success. Although cuterebrid bot flies are among the physically largest and most-studied insect parasites of mammals, the only study conducted on metabolic consequences of bot fly parasitism revealed a surprisingly small effect of bot flies on host metabolism. Here we test the prediction that bot fly parasitism increases the resting metabolic rate (RMR) of free-ranging eastern chipmunks (Tamias striatus), particularly in juveniles who have not previously encountered parasites and have to allocate energy to growth. We found no effect of bot fly parasitism on adults. In juveniles, however, we found that RMR strongly increased with the number of bot fly larvae hosted. For a subset of 12 juveniles during a year where parasite prevalence was particularly high, we also compared the RMR before versus during the peak of bot fly prevalence, allowing each individual to act as its own control. Each bot fly larva resulted in a approximately 7.6% increase in the RMR of its host while reducing juvenile growth rates. Finally, bot fly parasitism at the juvenile stage was positively correlated with adult stage RMR, suggesting persistent effects of bot flies on RMR. This study is the first to show an important effect of bot fly parasitism on the metabolism and growth of a wild mammal. Our work highlights the importance of studying cost of parasitism over multiple years in natural settings, as negative effects on hosts are more likely to emerge in periods of high energetic demand (e.g. growing juveniles) and/or in harsh environmental conditions (e.g. low food availability).
Can human-like Bots control collective mood: agent-based simulations of online chats
NASA Astrophysics Data System (ADS)
Tadić, Bosiljka; Šuvakov, Milovan
2013-10-01
Using an agent-based modeling approach, in this paper, we study self-organized dynamics of interacting agents in the presence of chat Bots. Different Bots with tunable ‘human-like’ attributes, which exchange emotional messages with agents, are considered, and the collective emotional behavior of agents is quantitatively analyzed. In particular, using detrended fractal analysis we determine persistent fluctuations and temporal correlations in time series of agent activity and statistics of avalanches carrying emotional messages of agents when Bots favoring positive/negative affects are active. We determine the impact of Bots and identify parameters that can modulate that impact. Our analysis suggests that, by these measures, the emotional Bots induce collective emotion among interacting agents by suitably altering the fractal characteristics of the underlying stochastic process. Positive emotion Bots are slightly more effective than negative emotion Bots. Moreover, Bots which periodically alternate between positive and negative emotion can enhance fluctuations in the system, leading to avalanches of agent messages that are reminiscent of self-organized critical states.
Fransen, Job; D'Hondt, Eva; Bourgois, Jan; Vaeyens, Roel; Philippaerts, Renaat M; Lenoir, Matthieu
2014-06-01
This study investigated convergent and discriminant validity between two motor competence assessment instruments in 2485 Flemish children: the Bruininks-Oseretsky Test of Motor Proficiency 2 Short Form (BOT-2 Short Form) and the KörperKoördinationsTest für Kinder (KTK). A Pearson correlation assessed the relationship between BOT-2 Short Form total, gross and fine motor composite scores and KTK Motor Quotient in three age cohorts (6-7, 8-9, 10-11 years). Crosstabs were used to measure agreement in classification in children scoring below percentile 5 and 15 and above percentile 85 and 95. Moderately strong positive (r=0.44-0.64) associations between BOT-2 total and gross motor composite scores and KTK Motor Quotient and weak positive correlations between BOT-2 Short Form fine motor composite and KTK Motor Quotient scores (r=0.25-0.37) were found. Levels of agreement were fair to moderate. Therefore, some proof of convergent and discriminant validity between BOT-2 Short Form and KTK was established in this study, underlining the notion that the evaluation of motor competence should not be based upon a single assessment instrument. Copyright © 2014 Elsevier Ltd. All rights reserved.
The botánica as a culturally appropriate health care option for Latinos.
Gomez-Beloz, A; Chavez, N
2001-10-01
The purpose of this study was to make a preliminary assessment of the botánica as a culturally appropriate health care option for Latinos in the United States. Closed-ended interviews were carried out using a questionnaire with 26 customers of a botánica located in a primarily urban Mexican American community in the United States. Sample gender was predominantly female with an average age of 38 years and income in the range of $15,000-$29,999 per year. Most participants were married (16), had children (17), and most (22) had formal education. Products purchased at the botánica included herbs, religious amulets, and other products. Although botánicas are commonly believed to provide remedies only for folk illnesses, the participants in this study reported seeking remedies primarily for somatic illnesses. The respondents used the services of conventional health care providers interchangeably with the botánica, making the botánica an important health care resource for Latinos in the United States. Further research should be carried out to better understand the role of botánicas in health care delivery.
Schäfer, Fabian; Evert, Stefan; Heinrich, Philipp
2017-12-01
In this article, we present results on the identification and behavioral analysis of social bots in a sample of 542,584 Tweets, collected before and after Japan's 2014 general election. Typical forms of bot activity include massive Retweeting and repeated posting of (nearly) the same message, sometimes used in combination. We focus on the second method and present (1) a case study on several patterns of bot activity, (2) methodological considerations on the automatic identification of such patterns and the prerequisite near-duplicate detection, and (3) we give qualitative insights into the purposes behind the usage of social/political bots. We argue that it was in the latency of the semi-public sphere of social media-and not in the visible or manifest public sphere (official campaign platform, mass media)-where Shinzō Abe's hidden nationalist agenda interlocked and overlapped with the one propagated by organizations such as Nippon Kaigi and Internet right-wingers (netto uyo) during the election campaign, the latter potentially forming an enormous online support army of Abe's agenda.
Schäfer, Fabian; Evert, Stefan; Heinrich, Philipp
2017-01-01
Abstract In this article, we present results on the identification and behavioral analysis of social bots in a sample of 542,584 Tweets, collected before and after Japan's 2014 general election. Typical forms of bot activity include massive Retweeting and repeated posting of (nearly) the same message, sometimes used in combination. We focus on the second method and present (1) a case study on several patterns of bot activity, (2) methodological considerations on the automatic identification of such patterns and the prerequisite near-duplicate detection, and (3) we give qualitative insights into the purposes behind the usage of social/political bots. We argue that it was in the latency of the semi-public sphere of social media—and not in the visible or manifest public sphere (official campaign platform, mass media)—where Shinzō Abe's hidden nationalist agenda interlocked and overlapped with the one propagated by organizations such as Nippon Kaigi and Internet right-wingers (netto uyo) during the election campaign, the latter potentially forming an enormous online support army of Abe's agenda. PMID:29182493
Braicu, Elena Ioana; Van Gorp, Toon; Nassir, Mani; Richter, Rolf; Chekerov, Radoslav; Gasimli, Khayal; Timmerman, Dirk; Vergote, Ignace; Sehouli, Jalid
2014-01-01
Borderline tumors of the ovary (BOT) are a distinct entity of ovarian tumors, characterized by lack of stromal invasion. Recent studies postulated that the presence of invasive implants, incomplete staging, fertility sparing surgery and residual tumor after surgery are major prognostic factors for BOT. There are no biomarkers that can predict BOT or the presence of invasive implants. The aim of our study was to assess the value of CA125 and HE4 alone, or within ROMA score for detecting BOT, and for predicting the presence of invasive implants. Retrospective, monocentric study on 167 women diagnosed with BOT or benign ovarian masses. Serum HE4, CA125 levels and ROMA were assessed preoperatively. Due to low number of BOT with invasive implants, we performed an unmatched analysis (consecutive patients) and a matched analysis (according to age and histology) to compare BOT with invasive implants, BOT without invasive implants and benign disease. There were no significant differences in the HE4 and CA125 expressions in the three groups of patients (p = 0.984 and p = 0.141, respectively). The ROC analysis showed that CA125 alone is superior to ROMA and HE4 in discriminating patients with BOT with invasive implants from patients with benign diseases and BOT without invasive implants. A newly established score, ROMABOT, did not perform better than ROMA. The analysis of the matched groups revealed similar results as the analysis of all samples. Both HE4 and CA125 are not reliable biomarkers for the diagnosis of BOT or for predicting the presence of invasive implants.
E-Cigarette Surveillance With Social Media Data: Social Bots, Emerging Topics, and Trends
Ferrara, Emilio; Uppu, Sree Priyanka; Cruz, Tess Boley; Unger, Jennifer B
2017-01-01
Background As e-cigarette use rapidly increases in popularity, data from online social systems (Twitter, Instagram, Google Web Search) can be used to capture and describe the social and environmental context in which individuals use, perceive, and are marketed this tobacco product. Social media data may serve as a massive focus group where people organically discuss e-cigarettes unprimed by a researcher, without instrument bias, captured in near real time and at low costs. Objective This study documents e-cigarette–related discussions on Twitter, describing themes of conversations and locations where Twitter users often discuss e-cigarettes, to identify priority areas for e-cigarette education campaigns. Additionally, this study demonstrates the importance of distinguishing between social bots and human users when attempting to understand public health–related behaviors and attitudes. Methods E-cigarette–related posts on Twitter (N=6,185,153) were collected from December 24, 2016, to April 21, 2017. Techniques drawn from network science were used to determine discussions of e-cigarettes by describing which hashtags co-occur (concept clusters) in a Twitter network. Posts and metadata were used to describe where geographically e-cigarette–related discussions in the United States occurred. Machine learning models were used to distinguish between Twitter posts reflecting attitudes and behaviors of genuine human users from those of social bots. Odds ratios were computed from 2x2 contingency tables to detect if hashtags varied by source (social bot vs human user) using the Fisher exact test to determine statistical significance. Results Clusters found in the corpus of hashtags from human users included behaviors (eg, #vaping), vaping identity (eg, #vapelife), and vaping community (eg, #vapenation). Additional clusters included products (eg, #eliquids), dual tobacco use (eg, #hookah), and polysubstance use (eg, #marijuana). Clusters found in the corpus of hashtags
E-Cigarette Surveillance With Social Media Data: Social Bots, Emerging Topics, and Trends.
Allem, Jon-Patrick; Ferrara, Emilio; Uppu, Sree Priyanka; Cruz, Tess Boley; Unger, Jennifer B
2017-12-20
As e-cigarette use rapidly increases in popularity, data from online social systems (Twitter, Instagram, Google Web Search) can be used to capture and describe the social and environmental context in which individuals use, perceive, and are marketed this tobacco product. Social media data may serve as a massive focus group where people organically discuss e-cigarettes unprimed by a researcher, without instrument bias, captured in near real time and at low costs. This study documents e-cigarette-related discussions on Twitter, describing themes of conversations and locations where Twitter users often discuss e-cigarettes, to identify priority areas for e-cigarette education campaigns. Additionally, this study demonstrates the importance of distinguishing between social bots and human users when attempting to understand public health-related behaviors and attitudes. E-cigarette-related posts on Twitter (N=6,185,153) were collected from December 24, 2016, to April 21, 2017. Techniques drawn from network science were used to determine discussions of e-cigarettes by describing which hashtags co-occur (concept clusters) in a Twitter network. Posts and metadata were used to describe where geographically e-cigarette-related discussions in the United States occurred. Machine learning models were used to distinguish between Twitter posts reflecting attitudes and behaviors of genuine human users from those of social bots. Odds ratios were computed from 2x2 contingency tables to detect if hashtags varied by source (social bot vs human user) using the Fisher exact test to determine statistical significance. Clusters found in the corpus of hashtags from human users included behaviors (eg, #vaping), vaping identity (eg, #vapelife), and vaping community (eg, #vapenation). Additional clusters included products (eg, #eliquids), dual tobacco use (eg, #hookah), and polysubstance use (eg, #marijuana). Clusters found in the corpus of hashtags from social bots included health (eg
Hanson-Smith, Victor; Johnson, Alexander
2016-07-01
The method of phylogenetic ancestral sequence reconstruction is a powerful approach for studying evolutionary relationships among protein sequence, structure, and function. In particular, this approach allows investigators to (1) reconstruct and "resurrect" (that is, synthesize in vivo or in vitro) extinct proteins to study how they differ from modern proteins, (2) identify key amino acid changes that, over evolutionary timescales, have altered the function of the protein, and (3) order historical events in the evolution of protein function. Widespread use of this approach has been slow among molecular biologists, in part because the methods require significant computational expertise. Here we present PhyloBot, a web-based software tool that makes ancestral sequence reconstruction easy. Designed for non-experts, it integrates all the necessary software into a single user interface. Additionally, PhyloBot provides interactive tools to explore evolutionary trajectories between ancestors, enabling the rapid generation of hypotheses that can be tested using genetic or biochemical approaches. Early versions of this software were used in previous studies to discover genetic mechanisms underlying the functions of diverse protein families, including V-ATPase ion pumps, DNA-binding transcription regulators, and serine/threonine protein kinases. PhyloBot runs in a web browser, and is available at the following URL: http://www.phylobot.com. The software is implemented in Python using the Django web framework, and runs on elastic cloud computing resources from Amazon Web Services. Users can create and submit jobs on our free server (at the URL listed above), or use our open-source code to launch their own PhyloBot server.
Hanson-Smith, Victor; Johnson, Alexander
2016-01-01
The method of phylogenetic ancestral sequence reconstruction is a powerful approach for studying evolutionary relationships among protein sequence, structure, and function. In particular, this approach allows investigators to (1) reconstruct and “resurrect” (that is, synthesize in vivo or in vitro) extinct proteins to study how they differ from modern proteins, (2) identify key amino acid changes that, over evolutionary timescales, have altered the function of the protein, and (3) order historical events in the evolution of protein function. Widespread use of this approach has been slow among molecular biologists, in part because the methods require significant computational expertise. Here we present PhyloBot, a web-based software tool that makes ancestral sequence reconstruction easy. Designed for non-experts, it integrates all the necessary software into a single user interface. Additionally, PhyloBot provides interactive tools to explore evolutionary trajectories between ancestors, enabling the rapid generation of hypotheses that can be tested using genetic or biochemical approaches. Early versions of this software were used in previous studies to discover genetic mechanisms underlying the functions of diverse protein families, including V-ATPase ion pumps, DNA-binding transcription regulators, and serine/threonine protein kinases. PhyloBot runs in a web browser, and is available at the following URL: http://www.phylobot.com. The software is implemented in Python using the Django web framework, and runs on elastic cloud computing resources from Amazon Web Services. Users can create and submit jobs on our free server (at the URL listed above), or use our open-source code to launch their own PhyloBot server. PMID:27472806
Bot, Cyborg and Automated Turing Test
NASA Astrophysics Data System (ADS)
Yan, Jeff
Ross Anderson: Bot tending might be an attractive activity for children, because children could receive the challenges on their mobile phones, to which they are almost physiologically attached these days, and they’re perhaps used to relatively smaller amounts of pocket money.
Bot fly parasitism of the red-backed vole: host survival, infection risk, and population growth.
Lemaître, Jérôme; Fortin, Daniel; Montiglio, Pierre-Olivier; Darveau, Marcel
2009-03-01
Parasites can play an important role in the dynamics of host populations, but empirical evidence remains sparse. We investigated the role of bot fly (Cuterebra spp.) parasitism in red-backed voles (Myodes gapperi) by first assessing the impacts of the parasite on the probability of vole survival under stressful conditions as well as on the reproductive activity of females. We then identified the main factors driving both the individual risk of infection and the abundance of bot flies inside red-backed voles. Finally, we evaluated the impacts of bot fly prevalence on the growth rate of vole populations between mid-July and mid-August. Thirty-six populations of red-backed voles were sampled in the boreal forest of Québec, Canada. The presence and the abundance of parasites in voles, two host life history traits (sex and body condition), three indices of habitat complexity (tree basal area, sapling basal area, coarse woody debris volume), and vole abundance were considered in models evaluating the effects of bot flies on host populations. We found that the probability of survival of red-backed voles in live traps decreased with bot fly infection. Both the individual risk of infection and the abundance of bot flies in red-backed voles were driven mainly by vole abundance rather than by the two host life history traits or the three variables of habitat complexity. Parasitism had population consequences: bot fly prevalence was linked to a decrease in short-term growth rate of vole populations over the summer. We found that bot flies have the potential to reduce survival of red-backed voles, an effect that may apply to large portions of populations.
The Battling 'Bots of Bloomsburg High
ERIC Educational Resources Information Center
Gorman, Lynn
2007-01-01
In this article, the author describes how students in Kirk Marshall's industrial technology class at Bloomsburg Area High School, Pennsylvania, designed and manufactured battling robots (BattleBots) and their participation in an annual national robotics competition. According to Marshall, designing and building a complex robot would be virtually…
The automatic neutron guide optimizer guide_bot
NASA Astrophysics Data System (ADS)
Bertelsen, Mads
2017-09-01
The guide optimization software guide_bot is introduced, the main purpose of which is to reduce the time spent programming when performing numerical optimization of neutron guides. A limited amount of information on the overall guide geometry and a figure of merit describing the desired beam is used to generate the code necessary to solve the problem. A generated McStas instrument file performs the Monte Carlo ray-tracing, which is controlled by iFit optimization scripts. The resulting optimal guide is thoroughly characterized, both in terms of brilliance transfer from an idealized source and on a more realistic source such as the ESS Butterfly moderator. Basic MATLAB knowledge is required from the user, but no experience with McStas or iFit is necessary. This paper briefly describes how guide_bot is used and some important aspects of the code. A short validation against earlier work is performed which shows the expected agreement. In addition a scan over the vertical divergence requirement, where individual guide optimizations are performed for each corresponding figure of merit, provides valuable data on the consequences of this parameter. The guide_bot software package is best suited for the start of an instrument design project as it excels at comparing a large amount of different guide alternatives for a specific set of instrument requirements, but is still applicable in later stages as constraints can be used to optimize more specific guides.
An investigation of douching practices in the botánicas of the Bronx.
Anderson, Matthew R; McKee, Diane; Yukes, Jolene; Alvarez, Adelyn; Karasz, Alison
2008-01-01
Douching is a common practice in women and has been associated with adverse health outcomes. In order to explore douching products and practices we conducted qualitative interviews in ten botánicas (stores that provide healing and spiritual services to immigrant communities) located in New York City. We interviewed 15 people, 14 of whom were botánica owners and employees and ten of whom were women. We found that douching was not easily separated from the more holistic concerns of botánica customers involving health, well-being and spirituality. These issues included abortion, infertility, menopause, the prevention and treatment of infections, sexuality, cleanliness, hygiene and relationship issues. The vagina was seen as a sensitive, even vulnerable part of the body, not clearly distinguished from other female organs. A variety of products were used in the vagina in the form of creams, douches, suppositories, baths and herbal steaming of the urogenital area. Alum, an astringent, was used for the purposes of vaginal tightening to enhance sexual pleasure for the partner, to make the vagina 'younger', or to hide evidence of infidelity. Botánicas are part of a complex healing system with conceptual models different from those of allopathic medicine. These models may not be unique to the botánicas.
Vercruysse, Jean-Philippe; van Dinther, Joost J S; De Foer, Bert; Casselman, Jan; Somers, Thomas; Zarowski, Andrzej; Cremers, Cor C; Offeciers, Erwin
2016-07-01
To present the long-term surgical outcome of the bony mastoid and epitympanic obliteration technique with canal wall reconstruction (CWR-BOT) in adults with an unstable cavity after previous canal wall-down surgery for extensive cholesteatoma. Retrospective study. Therapeutic. Tertiary referral center. Fifty consecutive adult patients undergoing a CWR-BOT between 1998 and 2009. (A) Recurrence and residual rates of cholesteatoma, (B) postoperative hygienic status of the ear, including postoperative aspect of the tympanic membrane and external ear canal integrity (EAC), (C) functional outcome, and (D) long-term safety issues. (A) The percentage of ears remaining safe without recurrent or residual disease after CWR-BOT was 96% after a mean follow-up time of 101.8 months. Recurrent cholesteatoma occurred in 2% (n = 1) and a residual cholesteatoma was detected in 2% (n = 1) of the patients. (B) A safe dry, and trouble-free graft and selfcleaning EAC was achieved in 94%. (C) The postoperative hearing results showed a gain of 1.7 dB on pure-tone average air-conduction. (D) Nonecho planar diffusion-weighted imaging (non-EP DW magnetic resonance imaging) documented the residual (n = 1) and recurrent cholesteatoma (n = 1). The 1- and 5-year imaging follow-up revealed no other recurrent or residual disease. The CWR-BOT is a safe and very effective option for treatment of problematic unstable canal wall-down mastoid cavities, resulting in dry trouble-free ears.
Integration Telegram Bot on E-Complaint Applications in College
NASA Astrophysics Data System (ADS)
Rosid, M. A.; Rachmadany, A.; Multazam, M. T.; Nandiyanto, A. B. D.; Abdullah, A. G.; Widiaty, I.
2018-01-01
Internet of Things (IoT) has influenced human life where IoT internet connectivity extending from human-to-humans to human-to-machine or machine-to-machine. With this research field, it will be created a technology and concepts that allow humans to communicate with machines for a specific purpose. This research aimed to integrate between application service of the telegram sender with application of e-complaint at a college. With this application, users do not need to visit the Url of the E-compliant application; but, they can be accessed simply by submitting a complaint via Telegram, and then the complaint will be forwarded to the E-complaint Application. From the test results, e-complaint integration with Telegram Bot has been run in accordance with the design. Telegram Bot is made able to provide convenience to the user in this academician to submit a complaint, besides the telegram bot provides the user interaction with the usual interface used by people everyday on their smartphones. Thus, with this system, the complained work unit can immediately make improvements since all the complaints process can be delivered rapidly.
Experiences with a Barista Robot, FusionBot
NASA Astrophysics Data System (ADS)
Limbu, Dilip Kumar; Tan, Yeow Kee; Wong, Chern Yuen; Jiang, Ridong; Wu, Hengxin; Li, Liyuan; Kah, Eng Hoe; Yu, Xinguo; Li, Dong; Li, Haizhou
In this paper, we describe the implemented service robot, called FusionBot. The goal of this research is to explore and demonstrate the utility of an interactive service robot in a smart home environment, thereby improving the quality of human life. The robot has four main features: 1) speech recognition, 2) object recognition, 3) object grabbing and fetching and 4) communication with a smart coffee machine. Its software architecture employs a multimodal dialogue system that integrates different components, including spoken dialog system, vision understanding, navigation and smart device gateway. In the experiments conducted during the TechFest 2008 event, the FusionBot successfully demonstrated that it could autonomously serve coffee to visitors on their request. Preliminary survey results indicate that the robot has potential to not only aid in the general robotics but also contribute towards the long term goal of intelligent service robotics in smart home environment.
Trillsch, Fabian; Ruetzel, Jan David; Herwig, Uwe; Doerste, Ulrike; Woelber, Linn; Grimm, Donata; Choschzick, Matthias; Jaenicke, Fritz; Mahner, Sven
2013-07-09
Surgery is the cornerstone for clinical management of patients with borderline ovarian tumors (BOT). As these patients have an excellent overall prognosis, perioperative morbidity is the critical point for decision making when the treatment strategy is developed and the primary surgical approach is defined. Clinical and surgical parameters of patients undergoing surgery for primary BOT at our institutions between 1993 and 2008 were analyzed with regard to perioperative morbidity depending on the surgical approach (laparotomy vs. laparoscopy). A total of 105 patients were analyzed (44 with primary laparoscopy [42%], 61 with primary laparotomy [58%]). Complete surgical staging was achieved in 33 patients at primary surgical approach (31.4%) frequently leading to formal indication of re-staging procedures. Tumor rupture was significantly more frequent during laparoscopy compared to laparotomy (29.5% vs. 13.1%, p = 0.038) but no other intraoperative complications were seen in laparoscopic surgery in contrast to 7 of 61 laparotomies (0% vs. 11.5%, p = 0.020). Postoperative complication rates were similar in both groups (19.7% vs. 18.2%, p = 0.848). Irrespective of the surgical approach, surgical management of BOT has acceptable rates of perioperative complications and morbidity. Choice of initial surgical approach can therefore be made independent of complication-concerns. As the recently published large retrospective AGO ROBOT study observed similar oncologic outcome for both approaches, laparoscopy can be considered for staging of patients with BOT if this appears feasible. An algorithm for the surgical management of BOT patients has been developed.
The experiences of undergraduate nursing students with bots in Second LifeRTM
NASA Astrophysics Data System (ADS)
Rose, Lesele H.
As technology continues to transform education from the status quo of traditional lecture-style instruction to an interactive engaging learning experience, students' experiences within the learning environment continues to change as well. This dissertation addressed the need for continuing research in advancing implementation of technology in higher education. The purpose of this phenomenological study was to discover more about the experiences of undergraduate nursing students using standardized geriatric evaluation tools when interacting with scripted geriatric patient bots tools in a simulated instructional intake setting. Data was collected through a Demographics questionnaire, an Experiential questionnaire, and a Reflection questionnaire. Triangulation of data collection occurred through an automatically created log of the interactions with the two bots, and by an automatically recorded log of the participants' movements while in the simulated geriatric intake interview. The data analysis consisted of an iterative review of the questionnaires and the participants' logs in an effort to identify common themes, recurring comments, and issues which would benefit from further exploration. Findings revealed that the interactions with the bots were perceived as a valuable experience for the participants from the perspective of interacting with the Geriatric Evaluation Tools in the role of an intake nurse. Further research is indicated to explore instructional interactions with bots in effectively mastering the use of established Geriatric Evaluation Tools.
Lee, Hyun Gyu; Cho, Nam-Chul; Jeong, Ae Jin; Li, Yu-Chen; Rhie, Sung-Ja; Choi, Jung Sook; Lee, Kwang-Ho; Kim, Youngsoo; Kim, Yong-Nyun; Kim, Myoung-Hwan; Pae, Ae Nim; Ye, Sang-Kyu; Kim, Byung-Hak
2016-01-01
T-cell-mediated immune responses play an important role in body protection. However, aberrantly activated immune responses are responsible for inflammatory and autoimmune diseases. The regulation of pathologic immune responses may be a potential therapeutic strategy for the treatment of these diseases. Despite that multiple pharmacologic properties of benzoxathiole derivatives have been defined, the molecular mechanisms underlying these properties remain to be clarified. Here, we demonstrated the benzoxathiole derivative 2-cyclohexylimino-6-methyl-6,7-dihydro-5H-benzo[1,3]oxathiol-4-one (BOT-4-one) regulated immune responses and ameliorated experimentally induced inflammatory skin diseases both in vitro and in vivo. BOT-4-one inhibited the differentiation of CD4(+) T-cell subsets by regulating the expression and production of T-cell lineage-specific master transcription factors and cytokines and activating the signal transducer and activator of transcription proteins. In addition, BOT-4-one inhibited TCR-mediated Akt and NF-κB signaling. Topical application of BOT-4-one ameliorated experimentally induced inflammatory skin diseases in mice models such as 2,4,6-trinitrochlorobenzene-induced contact and atopic dermatitis and IL-23-induced psoriasis-like skin inflammation. Our study demonstrated that BOT-4-one ameliorates inflammatory skin diseases by suppressing the pathogenic CD4(+) T cell differentiation and overall immune responses. Copyright © 2015 The Authors. Published by Elsevier Inc. All rights reserved.
NASA Astrophysics Data System (ADS)
Sarkar, Jayanta; Puska, Antti; Hassel, Juha; Hakonen, Pertti
2014-03-01
Bloch oscillating transistor (BOT) is mesoscopic current amplier based on a combination of a Josephson junction or a squid connected with a large resistor and a NIS junction. We have studied the dynamics of BOT near the bifurcation threshold. This is an important feature for an amplifier as this can be utilized to improve its performance characteristics. We have measured the I - V characteristics of the BOT with different base currents (IB) over a wide range of Josephson coupling energies (EJ) . The current gain (β) is found to be increasing with increasing IB and eventually diverging. We have found a record large β = 50 in our experiment. In order to determine the common mode rejection ratio (CMRR) of a differential pair BOT we have used two BOTs fabricated on the same chip. The common mode port is connected to the bases of the two BOTs and fed with varying voltages; simultaneously emitter currents of the two BOTs are recorded. In our experiment we found a 20dB of CMRR.
Martin-Eauclaire, Marie-France; Salvatierra, Juan; Bosmans, Frank; Bougis, Pierre E
2016-09-01
We report the detailed chemical, immunological and pharmacological characterization of the α-toxin Bot IX from the Moroccan scorpion Buthus occitanus tunetanus venom. Bot IX, which consists of 70 amino acids, is a highly atypical toxin. It carries a unique N-terminal sequence extension and is highly lethal in mice. Voltage clamp recordings on oocytes expressing rat Nav1.2 or insect BgNav1 reveal that, similar to other α-like toxins, Bot IX inhibits fast inactivation of both variants. Moreover, Bot IX belongs to the same structural/immunological group as the α-like toxin Bot I. Remarkably, radioiodinated Bot IX competes efficiently with the classical α-toxin AaH II from Androctonus australis, and displays one of the highest affinities for Nav channels. © 2016 Federation of European Biochemical Societies.
Discriminating bot accounts based solely on temporal features of microblog behavior
NASA Astrophysics Data System (ADS)
Pan, Junshan; Liu, Ying; Liu, Xiang; Hu, Hanping
2016-05-01
As the largest microblog service in China, Sina Weibo has attracted numerous automated applications (known as bots) due to its popularity and open architecture. We classify the active users from Sina Weibo into human, bot-based and hybrid groups based solely on the study of temporal features of their posting behavior. The anomalous burstiness parameter and time-interval entropy value are exploited to characterize automation. We also reveal different behavior patterns among the three types of users regarding their reposting ratio, daily rhythm and active days. Our findings may help Sina Weibo manage a better community and should be considered for dynamic models of microblog behaviors.
Lee, Hyun Gyu; Cho, Nam-Chul; Jeong, Ae Jin; Li, Yu-Chen; Rhie, Sung-Ja; Choi, Jung Sook; Lee, Kwang-Ho; Kim, Youngsoo; Kim, Yong-Nyun; Kim, Myoung-Hwan; Pae, Ae Nim; Ye, Sang-Kyu; Kim, Byung-Hak
2015-09-30
T cell-mediated immune responses play an important role in body protection. However, aberrantly activated immune responses are responsible for inflammatory and autoimmune diseases. The regulation of pathological immune responses may be a potential therapeutic strategy for the treatment of these diseases. Despite multiple pharmacological properties of benzoxathiole derivatives have been defined, the molecular mechanisms underlying these properties remain to be clarified. Here, we demonstrated the benzoxathiole derivative 2-cyclohexylimino-6-methyl-6,7-dihydro-5H-benzo[1,3]oxathiol-4-one (BOT-4-one) regulated immune responses and ameliorated experimentally induced inflammatory skin diseases both in vitro and in vivo. BOT-4-one inhibited the differentiation of CD4 + T-cell subsets by regulating the expression and production of T cell lineage-specific master transcription factors and cytokines and activating the signal transducer and activator of transcription (STAT) proteins. In addition, BOT-4-one inhibited T-cell receptor (TCR)-mediated Akt and nuclear factor-kappaB (NF-κB) signaling. Topical application of BOT-4-one ameliorated experimentally induced inflammatory skin diseases in mice models such as TNCB-induced contact and atopic dermatitis and IL-23-induced psoriasis-like skin inflammation. Our study demonstrated that BOT-4-one ameliorates inflammatory skin diseases by suppressing the pathogenic CD4 + T cell differentiation and the overall immune responses.Journal of Investigative Dermatology accepted article preview online, 30 September 2015. doi:10.1038/jid.2015.384.
Suzuki, Daisuke; Umezono, Tomoya; Miyauchi, Masaaki; Kimura, Moritsugu; Yamamoto, Naoyuki; Tanaka, Eitaro; Kuriyama, Yusuke; Sato, Hiroki; Miyatake, Han; Kondo, Masumi; Toyoda, Masao; Fukagawa, Masafumi
2012-07-20
To determine the clinical usefulness of basal-supported oral therapy (BOT) using insulin glargine in Japanese patients with type 2 diabetes. We compared HbA1c levels, body weight, and insulin doses before the introduction of BOT and in the final month of the observation period in 122 patients with type 2 diabetes who received BOT with insulin glargine between October 2007 and July 2009. To exclude the possible effects of seasonal changes in glycemic control, 57 of the 122 patients were followed-up for one year and examined for changes in HbA1c levels, body weight, and insulin dose. Examination of all cases (n=122) showed a significant decrease in HbA1c (before BOT: 8.7±1.8, after: 7.1±1.1%), but no significant change in body weight (before: 63.1±16.1, after: 63.8±17.0 kg). The mean observation period was 10.5±6.4 months. Insulin doses were significantly increased during the study. HbA1c levels improved significantly in patients on non-insulin-secreting drugs (biguanide, α-glucosidase inhibitor and thiazolidine derivatives) than those on insulin-secreting drugs (SU agents and glinides). BOT with insulin glargine is a useful strategy that can achieve good glycemic control in clinical practice without causing serious hypoglycemia. The introduction of BOT before exhaustion of pancreatic β cells may increase its effectiveness.
Passive tracking scheme for a single stationary observer
NASA Astrophysics Data System (ADS)
Chan, Y. T.; Rea, Terry
2001-08-01
While there are many techniques for Bearings-Only Tracking (BOT) in the ocean environment, they do not apply directly to the land situation. Generally, for tactical reasons, the land observer platform is stationary; but, it has two sensors, visual and infrared, for measuring bearings and a laser range finder (LRF) for measuring range. There is a requirement to develop a new BOT data fusion scheme that fuses the two sets of bearing readings, and together with a single LRF measurement, produces a unique track. This paper first develops a parameterized solution for the target speeds, prior to the occurrence of the LRF measurement, when the problem is unobservable. At, and after, the LRF measurement, a BOT formulated as a least squares (LS) estimator then produces a unique LS estimate of the target states. Bearing readings from the other sensor serve as instrumental variables in a data fusion setting to eliminate the bias in the BOT estimator. The result is recursive, unbiased and decentralized data fusion scheme. Results from two simulation experiments have corroborated the theoretical development and show that the scheme is optimal.
NASA Astrophysics Data System (ADS)
Zeb Gul, Jahan; Yang, Bong-Su; Yang, Young Jin; Chang, Dong Eui; Choi, Kyung Hyun
2016-11-01
Soft bots have the expedient ability of adopting intricate postures and fitting in complex shapes compared to mechanical robots. This paper presents a unique in situ UV curing three-dimensional (3D) printed multi-material tri-legged soft bot with spider mimicked multi-step dynamic forward gait using commercial bio metal filament (BMF) as an actuator. The printed soft bot can produce controllable forward motion in response to external signals. The fundamental properties of BMF, including output force, contractions at different frequencies, initial loading rate, and displacement-rate are verified. The tri-pedal soft bot CAD model is designed inspired by spider’s legged structure and its locomotion is assessed by simulating strain and displacement using finite element analysis. A customized rotational multi-head 3D printing system assisted with multiple wavelength’s curing lasers is used for in situ fabrication of tri-pedal soft-bot using two flexible materials (epoxy and polyurethane) in three layered steps. The size of tri-pedal soft-bot is 80 mm in diameter and each pedal’s width and depth is 5 mm × 5 mm respectively. The maximum forward speed achieved is 2.7 mm s-1 @ 5 Hz with input voltage of 3 V and 250 mA on a smooth surface. The fabricated tri-pedal soft bot proved its power efficiency and controllable locomotion at three input signal frequencies (1, 2, 5 Hz).
Single-cell copy number variation detection
2011-01-01
Detection of chromosomal aberrations from a single cell by array comparative genomic hybridization (single-cell array CGH), instead of from a population of cells, is an emerging technique. However, such detection is challenging because of the genome artifacts and the DNA amplification process inherent to the single cell approach. Current normalization algorithms result in inaccurate aberration detection for single-cell data. We propose a normalization method based on channel, genome composition and recurrent genome artifact corrections. We demonstrate that the proposed channel clone normalization significantly improves the copy number variation detection in both simulated and real single-cell array CGH data. PMID:21854607
OzBot and haptics: remote surveillance to physical presence
NASA Astrophysics Data System (ADS)
Mullins, James; Fielding, Mick; Nahavandi, Saeid
2009-05-01
This paper reports on robotic and haptic technologies and capabilities developed for the law enforcement and defence community within Australia by the Centre for Intelligent Systems Research (CISR). The OzBot series of small and medium surveillance robots have been designed in Australia and evaluated by law enforcement and defence personnel to determine suitability and ruggedness in a variety of environments. Using custom developed digital electronics and featuring expandable data busses including RS485, I2C, RS232, video and Ethernet, the robots can be directly connected to many off the shelf payloads such as gas sensors, x-ray sources and camera systems including thermal and night vision. Differentiating the OzBot platform from its peers is its ability to be integrated directly with haptic technology or the 'haptic bubble' developed by CISR. Haptic interfaces allow an operator to physically 'feel' remote environments through position-force control and experience realistic force feedback. By adding the capability to remotely grasp an object, feel its weight, texture and other physical properties in real-time from the remote ground control unit, an operator's situational awareness is greatly improved through Haptic augmentation in an environment where remote-system feedback is often limited.
Shim, Do-Wan; Shin, Woo-Young; Yu, Sang-Hyeun; Kim, Byung-Hak; Ye, Sang-Kyu; Koppula, Sushruta; Won, Hyung-Sik; Kang, Tae-Bong; Lee, Kwang-Ho
2017-11-08
The ATPase activity of NLRP3 has pivotal role in inflammasome activation and is recognized as a good target for the development of the NLRP3 inflammasome-specific inhibitor. However, signals in the vicinity of the ATPase activity of NLRP3 have not been fully elucidated. Here, we demonstrate NLRP3 inflammasome-specific action of a benzoxathiole derivative, BOT-4-one. BOT-4-one exhibited an inhibition of NLRP3 inflammasome activation, which was attributable to its alkylating capability to NLRP3. In particular, the NLRP3 alkylation by BOT-4-one led to an impaired ATPase activity of NLRP3, thereby obstructing the assembly of the NLRP3 inflammasome. Additionally, we found that NLRP3 alkylators, including BOT-4-one, enhance the ubiquitination level of NLRP3, which might also contribute to the inhibition of NLRP3 inflammasome activation. Finally, BOT-4-one appeared to be superior to other known NLRP3 alkylators in inhibiting the functionality of the NLRP3 inflammasome and its resulting anti-inflammatory activity was confirmed in vivo using a monosodium urate-induced peritonitis mouse model. Collectively, the results suggest that NLRP3 alkylators function by inhibiting ATPase activity and increasing the ubiquitination level of NLRP3, and BOT-4-one could be the type of NLRP3 inhibitor that may be potentially useful for the novel development of a therapeutic agent in controlling NLRP3 inflammasome-related diseases.
NASA Technical Reports Server (NTRS)
Hanold, Gregg T.; Hanold, David T.
2010-01-01
This paper presents a new Route Generation Algorithm that accurately and realistically represents human route planning and navigation for Military Operations in Urban Terrain (MOUT). The accuracy of this algorithm in representing human behavior is measured using the Unreal Tournament(Trademark) 2004 (UT2004) Game Engine to provide the simulation environment in which the differences between the routes taken by the human player and those of a Synthetic Agent (BOT) executing the A-star algorithm and the new Route Generation Algorithm can be compared. The new Route Generation Algorithm computes the BOT route based on partial or incomplete knowledge received from the UT2004 game engine during game play. To allow BOT navigation to occur continuously throughout the game play with incomplete knowledge of the terrain, a spatial network model of the UT2004 MOUT terrain is captured and stored in an Oracle 11 9 Spatial Data Object (SOO). The SOO allows a partial data query to be executed to generate continuous route updates based on the terrain knowledge, and stored dynamic BOT, Player and environmental parameters returned by the query. The partial data query permits the dynamic adjustment of the planned routes by the Route Generation Algorithm based on the current state of the environment during a simulation. The dynamic nature of this algorithm more accurately allows the BOT to mimic the routes taken by the human executing under the same conditions thereby improving the realism of the BOT in a MOUT simulation environment.
Horn, Lars-Christian; Höhn, Anne K; Einenkel, Jens; Siebolts, Udo
2014-11-01
Molecular studies have shown that the most prevalent mutations in serous ovarian borderline tumors (s-BOT) are BRAF and/or KRAS alterations. About one third of s-BOT represent peritoneal implants and/or lymph node involvement. These extraovarian deposits may be monoclonal or polyclonal in origin. To test both the hypotheses, mutational analyses using pyrosequencing for BRAF codon 600 and KRAS codon 12/13 and 61 of microdissected tissue was performed in 15 s-BOT and their invasive and noninvasive peritoneal implants. Two to 6 implants from different peritoneal sites were examined in 13 cases. Lymph node deposits were available for the analysis in 3 cases. Six s-BOT showed mutation in exon 2 codon 12 of the KRAS proto-oncogen. Five additional cases showed BRAF p.V600E mutation representing an overall mutation rate of 73.3%. Multiple (2-6) peritoneal implants were analyzed after microdissection in 13 of 15 cases. All showed identical mutational results when compared with the ovarian site of the disease. All lymph node deposits, including those with multiple deposits in different nodes, showed identical results, suggesting high intratumoral mutational homogeneity. The evidence presented in this study and the majority of data reported in the literature support the hypothesis that s-BOT with their peritoneal implants and lymph node deposits show identical mutational status of BRAF and KRAS suggesting a monoclonal rather than a polyclonal disease regarding these both tested genetic loci. In addition, a high intratumoral genetic homogeneity can be suggested. In conclusion, the results of the present study support the monoclonal origin of s-BOT and their peritoneal implants and lymph node deposits.
Beam On Target (BOT) Produces Gamma Ray Burst (GRB) Fireballs and Afterglows
NASA Astrophysics Data System (ADS)
Greyber, H. D.
1997-12-01
Unlike the myriads of ad hoc models that have been offered to explain GRB, the BOT process is simply the very common process used worldwide in accelerator laboratories to produce gamma rays. The Strong Magnetic Field (SMF) model postulates an extremely intense, highly relativistic current ring formed during the original gravitational collapse of a distant galaxy when the plasma cloud was permeated by a primordial magnetic field. GRB occur when solid matter (asteroid, white dwarf, neutron star, planet) falls rapidly through the Storage Ring beam producing a very strongly collimated electromagnetic shower, and a huge amount of matter from the target, in the form of a giant, hot, expanding plasma cloud, or ``Fireball,'' is blown off. BOT satisfies all the ``severe constraints imposed on the source of this burst --'' concluded by the CGRO team (Sommer et al, Astrophys. J. 422 L63 (1994)) for the huge intense burst GRB930131, whereas neutron star merger models are ``difficult to reconcile.'' BOT expects the lowest energy gamma photons to arrive very slightly later than higher energy photons due to the time for the shower to penetrate the target. The millisecond spikes in bursts are due to the slender filaments of current that make up the Storage Ring beam. Delayed photons can be explained by a broken target ``rock.'' See H. Greyber in the book ``Compton Gamma Ray Observatory,'' AIP Conf. Proc. 280, 569 (1993).
Olah, George; Vigo, Gabriela; Ortiz, Lizzie; Rozsa, Lajos; Brightsmith, Donald J
2013-09-01
Bot fly larvae (Philornis genus) are obligate subcutaneous blood-feeding parasites of Neotropical birds including psittacines. We analyze twelve years of data on scarlet macaw (Ara macao) nestlings in natural and artificial nests in the lowland forests of southeastern Peru and report prevalence and intensity of Philornis parasitism. Bot fly prevalence was 28.9% while mean intensity was 5.0 larvae per infected chick. Prevalence in natural nests (11%, N=90 nestlings) was lower than in wooden nest-boxes (39%, N=57) and PVC boxes (39%, N=109). We describe a new technique of removing Philornis larvae using a reverse syringe design snake bite extractor. We compare this new technique to two other methods for removing bots from macaw chicks and find the new method the most suitable. Copyright © 2013 Elsevier B.V. All rights reserved.
The White-hat Bot: A Novel Botnet Defense Strategy
2012-06-14
etc. I will briefly discuss one common exploit here. One fraudulent activity 4 perpetuated by botnets involves ad services such as Google’s AdSense ...which pays website owners revenue for posting the AdSense banner on their web site (Google, 2012). The AdSense banner displays messages from...botmaster creates a bot that is programmed to visit the botmaster’s own websites to click on the advertisements displayed in the AdSense banners. Since
Five Years of the RoBOT "Rocks Beneath Our Toes" High School Outreach Program
NASA Astrophysics Data System (ADS)
Baxter, E. F.
2011-12-01
The "Rocks Beneath Our Toes" or RoBOT Program began in 2006 as part of an NSF CAREER award through the Geochemistry and Petrology Program. The educational outreach program engages Boston area high school students in a hands on study of rocks and minerals collected in their communities. The goal is to provide high school students a unique window into modern scientific methods of geochemistry and mineralogy and create a higher level of interest and awareness of geoscience amongst Massachusetts secondary school students who are less often exposed to earth science coursework. Beginning with a joint field trip to sampling sites identified by participants, high school students work with Boston University undergraduates enrolled in Mineralogy to analyze their samples in thin section. During the field trip, each BU undergraduate is paired with a high school student. The assignment of student pairings (started in year 2) dramatically increased student interactions and enjoyment. The program culminates with a visit by the high school group to tour BU's lab facilities and work with the undergraduates using the petrographic microscopes to explore their rock. At this visit, BU undergraduates present their semester's work in one-on-one powerpoint presentations from which discussion and microscope work follow. Thus far, >50 high school students, >40 undergraduates, and 7 high school educators were involved in the program. This included participants from three different suburban Boston area high schools and with students enrolled in the BU "Upward Bound" program: an existing program designed to enhance educational opportunities for Boston inner city high school students. Participant reviews indicate great success in achieving the program's goals. Notably, both BU undergraduates and high school students rated the opportunities for interaction with eachother among the best aspects of RoBOT. On a scale of 1 to 10, BU undergraduates rated the following four categories highest
NASA Astrophysics Data System (ADS)
Trauth, N.; Schmidt, C.; Munz, M.
2016-12-01
Heat as a natural tracer to quantify water fluxes between groundwater and surface water has evolved to a standard hydrological method. Typically, time series of temperatures in the surface water and in the sediment are observed and are subsequently evaluated by a vertical 1D representation of heat transport by advection and dispersion. Several analytical solutions as well as their implementation into user-friendly software exist in order to estimate water fluxes from the observed temperatures. Analytical solutions can be easily implemented but assumptions on the boundary conditions have to be made a priori, e.g. sinusoidal upper temperature boundary. Numerical models offer more flexibility and can handle temperature data which is characterized by irregular variations such as storm-event induced temperature changes and thus cannot readily be incorporated in analytical solutions. This also reduced the effort of data preprocessing such as the extraction of the diurnal temperature variation. We developed a software to estimate water FLUXes Based On Temperatures- FLUX-BOT. FLUX-BOT is a numerical code written in MATLAB which is intended to calculate vertical water fluxes in saturated sediments, based on the inversion of measured temperature time series observed at multiple depths. It applies a cell-centered Crank-Nicolson implicit finite difference scheme to solve the one-dimensional heat advection-conduction equation. Besides its core inverse numerical routines, FLUX-BOT includes functions visualizing the results and functions for performing uncertainty analysis. We provide applications of FLUX-BOT to generic as well as to measured temperature data to demonstrate its performance.
Kim, Byung Hak; Min, Yun Sook; Choi, Jung Sook; Baeg, Gyeong-Hun; Kim, Youngsoo; Shin, Jong Wook; Kim, Tae-Yoon
2011-01-01
Persistently activated JAK/STAT3 signaling pathway plays a pivotal role in various human cancers including major carcinomas and hematologic tumors, and is implicated in cancer cell survival and proliferation. Therefore, inhibition of JAK/STAT3 signaling may be a clinical application in cancer therapy. Here, we report that 2-cyclohexylimino-6-methyl-6,7-dihydro-5H-benzo [1,3]oxathiol-4-one (BOT-4-one), a small molecule inhibitor of JAK/STAT3 signaling, induces apoptosis through inhibition of STAT3 activation. BOT-4-one suppressed cytokine (upd)-induced tyrosine phosphorylation and transcriptional activity of STAT92E, the sole Drosophila STAT homolog. Consequently, BOT-4-one significantly inhibited STAT3 tyrosine phosphorylation and expression of STAT3 downstream target gene SOCS3 in various human cancer cell lines, and its effect was more potent in JAK3-activated Hodgkin's lymphoma cell line than in JAK2-activated breast cancer and prostate cancer cell lines. In addition, BOT-4-one-treated Hodgkin's lymphoma cells showed decreased cell survival and proliferation by inducing apoptosis through down-regulation of STAT3 downstream target anti-apoptotic gene expression. These results suggest that BOT-4-one is a novel small molecule inhibitor of JAK3/STAT3 signaling and may have therapeutic potential in the treatment of human cancers harboring aberrant JAK3/STAT3 signaling, specifically Hodgkin's lymphoma. PMID:21499010
Kim, Byung Hak; Min, Yun Sook; Choi, Jung Sook; Baeg, Gyeong Hun; Kim, Young Soo; Shin, Jong Wook; Kim, Tae Yoon; Ye, Sang Kyu
2011-05-31
Persistently activated JAK/STAT3 signaling pathway plays a pivotal role in various human cancers including major carcinomas and hematologic tumors, and is implicated in cancer cell survival and proliferation. Therefore, inhibition of JAK/STAT3 signaling may be a clinical application in cancer therapy. Here, we report that 2-cyclohexylimino-6-methyl-6,7-dihydro-5H-benzo [1,3]oxathiol-4-one (BOT-4-one), a small molecule inhibitor of JAK/STAT3 signaling, induces apoptosis through inhibition of STAT3 activation. BOT-4-one suppressed cytokine (upd)-induced tyrosine phosphorylation and transcriptional activity of STAT92E, the sole Drosophila STAT homolog. Consequently, BOT-4-one significantly inhibited STAT3 tyrosine phosphorylation and expression of STAT3 downstream target gene SOCS3 in various human cancer cell lines, and its effect was more potent in JAK3-activated Hodgkin's lymphoma cell line than in JAK2-activated breast cancer and prostate cancer cell lines. In addition, BOT-4-one-treated Hodgkin's lymphoma cells showed decreased cell survival and proliferation by inducing apoptosis through down-regulation of STAT3 downstream target anti-apoptotic gene expression. These results suggest that BOT-4-one is a novel small molecule inhibitor of JAK3/STAT3 signaling and may have therapeutic potential in the treatment of human cancers harboring aberrant JAK3/STAT3 signaling, specifically Hodgkin's lymphoma.
Combining single-molecule manipulation and single-molecule detection.
Cordova, Juan Carlos; Das, Dibyendu Kumar; Manning, Harris W; Lang, Matthew J
2014-10-01
Single molecule force manipulation combined with fluorescence techniques offers much promise in revealing mechanistic details of biomolecular machinery. Here, we review force-fluorescence microscopy, which combines the best features of manipulation and detection techniques. Three of the mainstay manipulation methods (optical traps, magnetic traps and atomic force microscopy) are discussed with respect to milestones in combination developments, in addition to highlight recent contributions to the field. An overview of additional strategies is discussed, including fluorescence based force sensors for force measurement in vivo. Armed with recent exciting demonstrations of this technology, the field of combined single-molecule manipulation and single-molecule detection is poised to provide unprecedented views of molecular machinery. Copyright © 2014 Elsevier Ltd. All rights reserved.
Trillsch, F; Mahner, S; Vettorazzi, E; Woelber, L; Reuss, A; Baumann, K; Keyver-Paik, M-D; Canzler, U; Wollschlaeger, K; Forner, D; Pfisterer, J; Schroeder, W; Muenstedt, K; Richter, B; Fotopoulou, C; Schmalfeldt, B; Burges, A; Ewald-Riegler, N; de Gregorio, N; Hilpert, F; Fehm, T; Meier, W; Hillemanns, P; Hanker, L; Hasenburg, A; Strauss, H-G; Hellriegel, M; Wimberger, P; Kommoss, S; Kommoss, F; Hauptmann, S; du Bois, A
2015-01-01
Background: Incomplete surgical staging is a negative prognostic factor for patients with borderline ovarian tumours (BOT). However, little is known about the prognostic impact of each individual staging procedure. Methods: Clinical parameters of 950 patients with BOT (confirmed by central reference pathology) treated between 1998 and 2008 at 24 German AGO centres were analysed. In 559 patients with serous BOT and adequate ovarian surgery, further recommended staging procedures (omentectomy, peritoneal biopsies, cytology) were evaluated applying Cox regression models with respect to progression-free survival (PFS). Results: For patients with one missing staging procedure, the hazard ratio (HR) for recurrence was 1.25 (95%-CI 0.66–2.39; P=0.497). This risk increased with each additional procedure skipped reaching statistical significance in case of two (HR 1.95; 95%-CI 1.06–3.58; P=0.031) and three missing steps (HR 2.37; 95%-CI 1.22–4.64; P=0.011). The most crucial procedure was omentectomy which retained a statistically significant impact on PFS in multiple analysis (HR 1.91; 95%-CI 1.15–3.19; P=0.013) adjusting for previously established prognostic factors as FIGO stage, tumour residuals, and fertility preservation. Conclusion: Individual surgical staging procedures contribute to the prognosis for patients with serous BOT. In this analysis, recurrence risk increased with each skipped surgical step. This should be considered when re-staging procedures following incomplete primary surgery are discussed. PMID:25562434
Trillsch, F; Mahner, S; Vettorazzi, E; Woelber, L; Reuss, A; Baumann, K; Keyver-Paik, M-D; Canzler, U; Wollschlaeger, K; Forner, D; Pfisterer, J; Schroeder, W; Muenstedt, K; Richter, B; Fotopoulou, C; Schmalfeldt, B; Burges, A; Ewald-Riegler, N; de Gregorio, N; Hilpert, F; Fehm, T; Meier, W; Hillemanns, P; Hanker, L; Hasenburg, A; Strauss, H-G; Hellriegel, M; Wimberger, P; Kommoss, S; Kommoss, F; Hauptmann, S; du Bois, A
2015-02-17
Incomplete surgical staging is a negative prognostic factor for patients with borderline ovarian tumours (BOT). However, little is known about the prognostic impact of each individual staging procedure. Clinical parameters of 950 patients with BOT (confirmed by central reference pathology) treated between 1998 and 2008 at 24 German AGO centres were analysed. In 559 patients with serous BOT and adequate ovarian surgery, further recommended staging procedures (omentectomy, peritoneal biopsies, cytology) were evaluated applying Cox regression models with respect to progression-free survival (PFS). For patients with one missing staging procedure, the hazard ratio (HR) for recurrence was 1.25 (95%-CI 0.66-2.39; P=0.497). This risk increased with each additional procedure skipped reaching statistical significance in case of two (HR 1.95; 95%-CI 1.06-3.58; P=0.031) and three missing steps (HR 2.37; 95%-CI 1.22-4.64; P=0.011). The most crucial procedure was omentectomy which retained a statistically significant impact on PFS in multiple analysis (HR 1.91; 95%-CI 1.15-3.19; P=0.013) adjusting for previously established prognostic factors as FIGO stage, tumour residuals, and fertility preservation. Individual surgical staging procedures contribute to the prognosis for patients with serous BOT. In this analysis, recurrence risk increased with each skipped surgical step. This should be considered when re-staging procedures following incomplete primary surgery are discussed.
Nakib, Imene; Martin-Eauclaire, Marie-France; Laraba-Djebari, Fatima
2016-10-01
Bot III neurotoxin is the most lethal α neurotoxin purified from Buthus occitanus tunetanus scorpion venom. This toxin binds to the voltage-gated sodium channel of excitable cells and blocks its inactivation, inducing an increased release of neurotransmitters (acetylcholine and catecholamines). This study aims to elucidate the involvement of cholinergic and adrenergic receptors in pathogenesis and inflammatory response triggered by this toxin. Injection of Bot III to animals induces an increase of peroxidase activities, an imbalance of oxidative status, tissue damages in lung parenchyma, and myocardium correlated with metabolic disorders. The pretreatment with nicotine (nicotinic receptor agonist) or atropine (muscarinic receptor antagonist) protected the animals from almost all disorders caused by Bot III toxin, especially the immunological alterations. Bisoprolol administration (selective β1 adrenergic receptor antagonist) was also efficient in the protection of animals, mainly on tissue damage. Propranolol (non-selective adrenergic receptor antagonist) showed less effect. These results suggest that both cholinergic and adrenergic receptors are activated in the cardiopulmonary manifestations induced by Bot III. Indeed, the muscarinic receptor appears to be more involved than the nicotinic one, and the β1 adrenergic receptor seems to dominate the β2 receptor. These results showed also that the activation of nicotinic receptor leads to a significant protection of animals against Bot III toxin effect. These findings supply a supplementary data leading to better understanding of the mechanism triggered by scorpionic neurotoxins and suggest the use of drugs targeting these receptors, especially the nicotinic one in order to counteract the inflammatory response observed in scorpion envenomation.
Electrical detection of single viruses
NASA Astrophysics Data System (ADS)
Patolsky, Fernando; Zheng, Gengfeng; Hayden, Oliver; Lakadamyali, Melike; Zhuang, Xiaowei; Lieber, Charles M.
2004-09-01
We report direct, real-time electrical detection of single virus particles with high selectivity by using nanowire field effect transistors. Measurements made with nanowire arrays modified with antibodies for influenza A showed discrete conductance changes characteristic of binding and unbinding in the presence of influenza A but not paramyxovirus or adenovirus. Simultaneous electrical and optical measurements using fluorescently labeled influenza A were used to demonstrate conclusively that the conductance changes correspond to binding/unbinding of single viruses at the surface of nanowire devices. pH-dependent studies further show that the detection mechanism is caused by a field effect, and that the nanowire devices can be used to determine rapidly isoelectric points and variations in receptor-virus binding kinetics for different conditions. Lastly, studies of nanowire devices modified with antibodies specific for either influenza or adenovirus show that multiple viruses can be selectively detected in parallel. The possibility of large-scale integration of these nanowire devices suggests potential for simultaneous detection of a large number of distinct viral threats at the single virus level.
Robotic upper limb rehabilitation after acute stroke by NeReBot: evaluation of treatment costs.
Stefano, Masiero; Patrizia, Poli; Mario, Armani; Ferlini, Gregorio; Rizzello, Roberto; Rosati, Giulio
2014-01-01
Stroke is the first cause of disability. Several robotic devices have been developed for stroke rehabilitation. Robot therapy by NeReBot is demonstrated to be an effective tool for the treatment of poststroke paretic upper limbs, able to improve the activities of daily living of stroke survivors when used both as additional treatment and in partial substitution of conventional rehabilitation therapy in the acute and subacute phases poststroke. This study presents the evaluation of the costs related to delivering such therapy, in comparison with conventional rehabilitation treatment. By comparing several NeReBot treatment protocols, made of different combinations of robotic and nonrobotic exercises, we show that robotic technology can be a valuable and economically sustainable aid in the management of poststroke patient rehabilitation.
Leitner, Natascha; Schwarzmann, Laurin; Zittra, Carina; Palmieri, Nicola; Eigner, Barbara; Otranto, Domenico; Glawischnig, Walter; Fuehrer, Hans-Peter
2016-11-01
Nasopharyngeal myiases are caused by larvae of bot flies (Diptera: Oestridae), which have evolved a high specificity for their hosts. Bot flies (n = 916) were collected from 137 (57.6 %) out of 238 red deer (Cervus elaphus) hunted in Vorarlberg and Tyrol (Western Austria). After being stored in 75 % ethanol, larvae were identified to species level and developmental stage using morphological and morphometric keys. Larvae were also molecularly characterized by polymerase chain reaction (PCR) amplification and partial sequencing of the mitochondrial cytochrome oxidase subunit I gene. Morphological and molecular analysis allowed identification of larvae as Cephenemyia auribarbis and Pharyngomyia picta. Genetic variations were also examined within the specimens collected in both geographical locations.
Remote detection of single emitters via optical waveguides
NASA Astrophysics Data System (ADS)
Then, Patrick; Razinskas, Gary; Feichtner, Thorsten; Haas, Philippe; Wild, Andreas; Bellini, Nicola; Osellame, Roberto; Cerullo, Giulio; Hecht, Bert
2014-05-01
The integration of lab-on-a-chip technologies with single-molecule detection techniques may enable new applications in analytical chemistry, biotechnology, and medicine. We describe a method based on the reciprocity theorem of electromagnetic theory to determine and optimize the detection efficiency of photons emitted by single quantum emitters through truncated dielectric waveguides of arbitrary shape positioned in their proximity. We demonstrate experimentally that detection of single quantum emitters via such waveguides is possible, confirming the predicted behavior of the detection efficiency. Our findings blaze the trail towards efficient lensless single-emitter detection compatible with large-scale optofluidic integration.
Quinzler, Renate; Ude, Miriam; Franzmann, Alexandra; Feldt, Sandra; Schüssel, Katrin; Leuner, Kristina; Müller, Walter E; Dippel, Franz-Werner; Schulz, Martin
2012-01-01
To compare the persistence (treatment duration) of basal insulin supported oral therapy (BOT) using insulin glargine (GLA) or NPH insulin (NPH) in Type-2 diabetic patients. This retrospective cohort study reports results from an analysis of claims data from prescriptions for ambulatory patients within the German Statutory Health Insurance scheme. The study is based on claims data from more than 80% of German community pharmacies. Treatment duration until switching to a basal bolus treatment regimen (intensified conventional insulin therapy: ICT) was determined in insulin-naïve patients who began treatment with BOT using GLA or NPH between 01/2003 and 12/2006. A total of 97,998 patients (61,070 GLA and 36,928 NPH) were included. Within the observation period, 23.5% of GLA patients and 28.0% of NPH patients switched from BOT to ICT. The upper quartile of probability of continuation of therapy (the 75th percentile) was reached after 769 days in GLA patients and after 517 days in NPH patients. Therefore, the risk of switching to ICT was significantly higher with NPH compared to GLA: hazard ratios were 1.34 (99% CI: 1.29-1.38; unadjusted) and 1.22 (99% CI: 1.18-1.27) after adjustment for predefined covariates. Various sensitivity analyses using modified inclusion criteria and endpoint definitions were applied and these confirmed the initial results. Type-2 diabetic patients under BOT with GLA stayed significantly longer on the initial therapy before switching to ICT than patients on BOT using NPH.
Training Knowledge Bots for Physics-Based Simulations Using Artificial Neural Networks
NASA Technical Reports Server (NTRS)
Samareh, Jamshid A.; Wong, Jay Ming
2014-01-01
Millions of complex physics-based simulations are required for design of an aerospace vehicle. These simulations are usually performed by highly trained and skilled analysts, who execute, monitor, and steer each simulation. Analysts rely heavily on their broad experience that may have taken 20-30 years to accumulate. In addition, the simulation software is complex in nature, requiring significant computational resources. Simulations of system of systems become even more complex and are beyond human capacity to effectively learn their behavior. IBM has developed machines that can learn and compete successfully with a chess grandmaster and most successful jeopardy contestants. These machines are capable of learning some complex problems much faster than humans can learn. In this paper, we propose using artificial neural network to train knowledge bots to identify the idiosyncrasies of simulation software and recognize patterns that can lead to successful simulations. We examine the use of knowledge bots for applications of computational fluid dynamics (CFD), trajectory analysis, commercial finite-element analysis software, and slosh propellant dynamics. We will show that machine learning algorithms can be used to learn the idiosyncrasies of computational simulations and identify regions of instability without including any additional information about their mathematical form or applied discretization approaches.
Real-time incident detection using social media data.
DOT National Transportation Integrated Search
2016-05-09
The effectiveness of traditional incident detection is often limited by sparse sensor coverage, and reporting incidents to emergency response systems : is labor-intensive. This research project mines tweet texts to extract incident information on bot...
Rugg, Douglas; Ferrer, Luis Miguel; Sarasola, Patxi; Figueras, Luis; Lacasta, Delia; Liu, Bo; Bartram, David
2012-09-10
Cydectin(®) 2% LA Solution for Injection for Sheep (Pfizer Animal Health) is a long-acting (LA) formulation of moxidectin for the treatment and prevention of mixed infections of gastro-intestinal nematodes, respiratory nematodes and certain arthropod parasites in sheep. To evaluate the duration of persistent efficacy against nasal bots (Oestrus ovis), a natural exposure study was conducted in Spain during the summer of 2011. One hundred and twenty nasal bot-free, Rasa Aragonesa sheep were randomly allocated to eight groups of 15 animals each. On Day 0, four groups were treated at the recommended dose rate of 1 mg moxidectin/kg bodyweight. Four groups remained untreated as negative controls. All animals were held in nasal bot-proof housing except for exposure to natural challenge when one group of treated sheep and one of group of control animals were transferred to a local pasture at either 0-20, 20-40, 40-60, or 60-80 days after treatment. Following challenge, sheep were scored for clinical signs of bot infestation, necropsied and the heads sectioned for larval recovery. Nasal bot larvae were retrieved from 7 to 11 control sheep following each exposure period indicating that adult bots were active throughout the study. In the first challenge up to 20 days after treatment, when sheep were slaughtered immediately after exposure, the majority of larvae were first instar (L1) and only 3 of the 15 control sheep were infested with second instars (L2). There was 100% efficacy against L2 and 38.1% reduction in the number of live L1 in the treated sheep but mean counts were not significantly different between treatment and control groups (P ≥ 0.05). For the subsequent exposure periods 20-80 days after treatment (necropsies 7-9 days after challenge), 6-10 sheep were infested with L1 and 9-11 control sheep were infested with L2 and third instars (L3). There was negligible efficacy against L1, but treatment with moxidectin resulted in 100% control of L2 and L3. These
Kim, Byung-Hak; Yoon, Bo Ruem; Kim, Eun Kyoung; Noh, Kum Hee; Kwon, Sun-Ho; Yi, Eun Hee; Lee, Hyun Gyu; Choi, Jung Sook; Kang, Seong Wook; Park, In-Chul; Lee, Won-Woo; Ye, Sang-Kyu
2016-06-15
Autoimmune rheumatoid arthritis is characterized by chronic inflammation and hyperplasia in the synovial joints. Although the cause of rheumatoid arthritis is largely unknown, substantial evidence has supported the importance of immune cells and inflammatory cytokines in the initiation and progression of this disease. Herein, we demonstrated that the benzoxathiole derivative 2-cyclohexylimino-6-methyl-6,7-dihydro-5H-benzo[1,3]oxathiol-4-one (BOT-4-one) alleviated type II collagen-induced arthritis in a mouse model. The levels of pro-inflammatory cytokines are elevated in both human patients with rheumatoid arthritis and mice with collagen-induced arthritis. BOT-4-one treatment reduced the levels of pro-inflammatory cytokines in mice and endotoxin-stimulated macrophages. BOT-4-one treatment suppressed the polarization of Th1- and Th17-cell subsets by inhibiting the expression and production of their lineage-specific master transcription factors and cytokines, as well as activation of signal transducer and activator of transcription proteins. In addition, BOT-4-one inhibited mitogen-activated protein kinase and NF-kappaB signaling as well as the transcriptional activities and DNA-binding of transcription factors, including activator protein-1, cAMP response element-binding protein and NF-kappaB. Our results suggest that BOT-4-one may have therapeutic potential for the treatment of chronic inflammation associated with autoimmune rheumatoid arthritis. Copyright © 2016 Elsevier Inc. All rights reserved.
Single molecule detection, thermal fluctuation and life
YANAGIDA, Toshio; ISHII, Yoshiharu
2017-01-01
Single molecule detection has contributed to our understanding of the unique mechanisms of life. Unlike artificial man-made machines, biological molecular machines integrate thermal noises rather than avoid them. For example, single molecule detection has demonstrated that myosin motors undergo biased Brownian motion for stepwise movement and that single protein molecules spontaneously change their conformation, for switching to interactions with other proteins, in response to thermal fluctuation. Thus, molecular machines have flexibility and efficiency not seen in artificial machines. PMID:28190869
Single-molecule detection: applications to ultrasensitive biochemical analysis
NASA Astrophysics Data System (ADS)
Castro, Alonso; Shera, E. Brooks
1995-06-01
Recent developments in laser-based detection of fluorescent molecules have made possible the implementation of very sensitive techniques for biochemical analysis. We present and discuss our experiments on the applications of our recently developed technique of single-molecule detection to the analysis of molecules of biological interest. These newly developed methods are capable of detecting and identifying biomolecules at the single-molecule level of sensitivity. In one case, identification is based on measuring fluorescence brightness from single molecules. In another, molecules are classified by determining their electrophoretic velocities.
Single DNA molecule detection using nanopipettes and nanoparticles.
Karhanek, Miloslav; Kemp, Jennifer T; Pourmand, Nader; Davis, Ronald W; Webb, Chris D
2005-02-01
Single DNA molecules labeled with nanoparticles can be detected by blockades of ionic current as they are translocated through a nanopipette tip formed by a pulled glass capillary. The nanopipette detection technique can provide not only tools for detection and identification of single DNA and protein molecules but also deeper insight and understanding of stochastic interactions of various biomolecules with their environment.
Hello Harlie: Enabling Speech Monitoring Through Chat-Bot Conversations.
Ireland, David; Atay, Christina; Liddle, Jacki; Bradford, Dana; Lee, Helen; Rushin, Olivia; Mullins, Thomas; Angus, Dan; Wiles, Janet; McBride, Simon; Vogel, Adam
2016-01-01
People with neurological conditions such as Parkinson's disease and dementia are known to have difficulties in language and communication. This paper presents initial testing of an artificial conversational agent, called Harlie. Harlie runs on a smartphone and is able to converse with the user on a variety of topics. A description of the application and a sample dialog are provided to illustrate the various roles chat-bots can play in the management of neurological conditions. Harlie can be used for measuring voice and communication outcomes during the daily life of the user, and for gaining information about challenges encountered. Moreover, it is anticipated that she may also have an educational and support role.
Atom-atom entanglement by single-photon detection.
Slodička, L; Hétet, G; Röck, N; Schindler, P; Hennrich, M; Blatt, R
2013-02-22
A scheme for entangling distant atoms is realized, as proposed in the seminal paper by [C. Cabrillo et al., Phys. Rev. A 59, 1025 (1999)]. The protocol is based on quantum interference and detection of a single photon scattered from two effectively one meter distant laser cooled and trapped atomic ions. The detection of a single photon heralds entanglement of two internal states of the trapped ions with high rate and with a fidelity limited mostly by atomic motion. Control of the entangled state phase is demonstrated by changing the path length of the single-photon interferometer.
The role of nanotechnology in single-cell detection: a review.
Wang, Changling; Zhang, Yuxiang; Xia, Mingdian; Zhu, Xingxi; Qi, Shitao; Shen, Huaqiang; Liu, Tiebing; Tang, Liming
2014-10-01
Biological processes in single cells, such as signal transduction, DNA duplication, and protein synthesis and trafficking, occur in subcellular compartments at nanoscale level. Achieving high spatial-temporal resolution, high sensitivity, and high specificity in single-cell detection poses a great challenge. Nanotechnology, which has been widely applied in the fields of medicine, electronics, biomaterials, and energy production, has the potential to provide solutions for single-cell detection. Here we present a review of the use of nanotechnology in single-cell detection over the past two decades. First, we review the main areas of scientific interest, including morphology, ion concentration, DNA, RNA, protein, intracellular temperature, elements, and mechanical properties. Second, four categories of application of nanotechnology to single-cell detection are described: nanomanipulation, nanodevices, nanomaterials as labels, and nano Secondary ion mass spectrometry. Finally, the prospects and future trends in single-cell detection and analysis are discussed.
Agent Models for Self-Motivated Home-Assistant Bots
NASA Astrophysics Data System (ADS)
Merrick, Kathryn; Shafi, Kamran
2010-01-01
Modern society increasingly relies on technology to support everyday activities. In the past, this technology has focused on automation, using computer technology embedded in physical objects. More recently, there is an expectation that this technology will not just embed reactive automation, but also embed intelligent, proactive automation in the environment. That is, there is an emerging desire for novel technologies that can monitor, assist, inform or entertain when required, and not just when requested. This paper presents three self-motivated, home-assistant bot applications using different self-motivated agent models. Self-motivated agents use a computational model of motivation to generate goals proactively. Technologies based on self-motivated agents can thus respond autonomously and proactively to stimuli from their environment. Three prototypes of different self-motivated agent models, using different computational models of motivation, are described to demonstrate these concepts.
Compositions and methods for detecting single nucleotide polymorphisms
Yeh, Hsin-Chih; Werner, James; Martinez, Jennifer S.
2016-11-22
Described herein are nucleic acid based probes and methods for discriminating and detecting single nucleotide variants in nucleic acid molecules (e.g., DNA). The methods include use of a pair of probes can be used to detect and identify polymorphisms, for example single nucleotide polymorphism in DNA. The pair of probes emit a different fluorescent wavelength of light depending on the association and alignment of the probes when hybridized to a target nucleic acid molecule. Each pair of probes is capable of discriminating at least two different nucleic acid molecules that differ by at least a single nucleotide difference. The methods can probes can be used, for example, for detection of DNA polymorphisms that are indicative of a particular disease or condition.
Self-referenced single-shot THz detection
DOE Office of Scientific and Technical Information (OSTI.GOV)
Russell, Brandon K.; Ofori-Okai, Benjamin K.; Chen, Zhijiang
We demonstrate a self-referencing method to reduce noise in a single-shot terahertz detection scheme. By splitting a single terahertz pulse and using a reflective echelon, both the signal and reference terahertz time-domain waveforms were measured using one laser pulse. Simultaneous acquisition of these waveforms significantly reduces noise originating from shot-to-shot fluctuations. Here, we show that correlation function based referencing, which is not limited to polarization dependent measurements, can achieve a noise floor that is comparable to state-of-the-art polarization-gated balanced detection. Lastly, we extract the DC conductivity of a 30 nm free-standing gold film using a single THz pulse. The measuredmore » value of σ 0 = 1.3 ± 0.4 × 10 7 S m -1 is in good agreement with the value measured by four-point probe, indicating the viability of this method for measuring dynamical changes and small signals.« less
Self-referenced single-shot THz detection
Russell, Brandon K.; Ofori-Okai, Benjamin K.; Chen, Zhijiang; ...
2017-06-29
We demonstrate a self-referencing method to reduce noise in a single-shot terahertz detection scheme. By splitting a single terahertz pulse and using a reflective echelon, both the signal and reference terahertz time-domain waveforms were measured using one laser pulse. Simultaneous acquisition of these waveforms significantly reduces noise originating from shot-to-shot fluctuations. Here, we show that correlation function based referencing, which is not limited to polarization dependent measurements, can achieve a noise floor that is comparable to state-of-the-art polarization-gated balanced detection. Lastly, we extract the DC conductivity of a 30 nm free-standing gold film using a single THz pulse. The measuredmore » value of σ 0 = 1.3 ± 0.4 × 10 7 S m -1 is in good agreement with the value measured by four-point probe, indicating the viability of this method for measuring dynamical changes and small signals.« less
Optofluidic wavelength division multiplexing for single-virus detection
Ozcelik, Damla; Parks, Joshua W.; Wall, Thomas A.; Stott, Matthew A.; Cai, Hong; Parks, Joseph W.; Hawkins, Aaron R.; Schmidt, Holger
2015-01-01
Optical waveguides simultaneously transport light at different colors, forming the basis of fiber-optic telecommunication networks that shuttle data in dozens of spectrally separated channels. Here, we reimagine this wavelength division multiplexing (WDM) paradigm in a novel context––the differentiated detection and identification of single influenza viruses on a chip. We use a single multimode interference (MMI) waveguide to create wavelength-dependent spot patterns across the entire visible spectrum and enable multiplexed single biomolecule detection on an optofluidic chip. Each target is identified by its time-dependent fluorescence signal without the need for spectral demultiplexing upon detection. We demonstrate detection of individual fluorescently labeled virus particles of three influenza A subtypes in two implementations: labeling of each virus using three different colors and two-color combinatorial labeling. By extending combinatorial multiplexing to three or more colors, MMI-based WDM provides the multiplexing power required for differentiated clinical tests and the growing field of personalized medicine. PMID:26438840
Warble? What’s a Warble? A recap of the human bot fly, Dermatobia hominis (L. Jr. 1781)
USDA-ARS?s Scientific Manuscript database
The human bot fly, Dermatobia hominis (Linnaeus Jr., 1781) is a major pest of livestock in Mexico, Central and South America. Myiasis caused by the larvae result in economic losses due to hide damage and reductions in weight gain and milk production. They have a broad host range which includes wildl...
Chen, Yen-Ting; Hsu, Chiao-Ling; Hou, Shao-Yi
2008-04-15
The current study reports an assay approach that can detect single-nucleotide polymorphisms (SNPs) and identify the position of the point mutation through a single-strand-specific nuclease reaction and a gold nanoparticle assembly. The assay can be implemented via three steps: a single-strand-specific nuclease reaction that allows the enzyme to truncate the mutant DNA; a purification step that uses capture probe-gold nanoparticles and centrifugation; and a hybridization reaction that induces detector probe-gold nanoparticles, capture probe-gold nanoparticles, and the target DNA to form large DNA-linked three-dimensional aggregates of gold nanoparticles. At high temperature (63 degrees C in the current case), the purple color of the perfect match solution would not change to red, whereas a mismatched solution becomes red as the assembled gold nanoparticles separate. Using melting analysis, the position of the point mutation could be identified. This assay provides a convenient colorimetric detection that enables point mutation identification without the need for expensive mass spectrometry. To our knowledge, this is the first report concerning SNP detection based on a single-strand-specific nuclease reaction and a gold nanoparticle assembly.
Single Bacterium Detection Using Sers
NASA Astrophysics Data System (ADS)
Gonchukov, S. A.; Baikova, T. V.; Alushin, M. V.; Svistunova, T. S.; Minaeva, S. A.; Ionin, A. A.; Kudryashov, S. I.; Saraeva, I. N.; Zayarny, D. A.
2016-02-01
This work is devoted to the study of a single Staphylococcus aureus bacterium detection using surface-enhanced Raman spectroscopy (SERS) and resonant Raman spectroscopy (RS). It was shown that SERS allows increasing sensitivity of predominantly low frequency lines connected with the vibrations of Amide, Proteins and DNA. At the same time the lines of carotenoids inherent to this kind of bacterium are well-detected due to the resonance Raman scattering mechanism. The reproducibility and stability of Raman spectra strongly depend on the characteristics of nanostructured substrate, and molecular structure and size of the tested biological object.
High sensitivity fluorescent single particle and single molecule detection apparatus and method
Mathies, Richard A.; Peck, Konan; Stryer, Lubert
1990-01-01
Apparatus is described for ultrasensitive detection of single fluorescent particles down to the single fluorescent molecule limit in a fluid or on a substrate comprising means for illuminating a predetermined volume of the fluid or area of the substrate whereby to emit light including background light from the fluid and burst of photons from particles residing in the area. The photon burst is detected in real time to generate output representative signal. The signal is received and the burst of energy from the fluorescent particles is distinguished from the background energy to provide an indication of the number, location or concentration of the particles or molecules.
Droplet microfluidics for amplification-free genetic detection of single cells.
Rane, Tushar D; Zec, Helena C; Puleo, Chris; Lee, Abraham P; Wang, Tza-Huei
2012-09-21
In this article we present a novel droplet microfluidic chip enabling amplification-free detection of single pathogenic cells. The device streamlines multiple functionalities to carry out sample digitization, cell lysis, probe-target hybridization for subsequent fluorescent detection. A peptide nucleic acid fluorescence resonance energy transfer probe (PNA beacon) is used to detect 16S rRNA present in pathogenic cells. Initially the sensitivity and quantification abilities of the platform are tested using a synthetic target mimicking the actual expression level of 16S rRNA in single cells. The capability of the device to perform "sample-to-answer" pathogen detection of single cells is demonstrated using E. coli as a model pathogen.
Treb-Bot: Development and Use of a Trebuchet Simulator
NASA Astrophysics Data System (ADS)
Constans, Eric; Constans, Aileen
2015-09-01
The trebuchet has quickly become a favorite project for physics and engineering teachers seeking to provide students with a simple, but spectacular, hands-on design project that can be applied to the study of projectile motion, rotational motion, and the law of conservation of energy. While there have been free trebuchet simulators and range calculators available online for several years, these have been limited to simple designs. Other simulators are available for a fee, precluding practical use in introductory courses. With this in mind, one of the authors developed a free web-based trebuchet simulation that can be found at http://www.benchtophybrid.com/TB_index.html. This simulation, named Treb-Bot, is designed to be visually appealing to high school students and includes simulations of trebuchet designs that are unavailable elsewhere on the web. The website was successfully field-tested by a group of Advanced Placement Physics 1 students.
Trillsch, F; Mahner, S; Woelber, L; Vettorazzi, E; Reuss, A; Ewald-Riegler, N; de Gregorio, N; Fotopoulou, C; Schmalfeldt, B; Burges, A; Hilpert, F; Fehm, T; Meier, W; Hillemanns, P; Hanker, L; Hasenburg, A; Strauss, H G; Hellriegel, M; Wimberger, P; Baumann, K; Keyver-Paik, M D; Canzler, U; Wollschlaeger, K; Forner, D; Pfisterer, J; Schroeder, W; Muenstedt, K; Richter, B; Kommoss, F; Hauptmann, S; du Bois, A
2014-07-01
Approximately one-third of all borderline ovarian tumours (BOT) are diagnosed in patients with child-bearing potential. Detailed information regarding their specific characteristics and prognostic factors is limited. Clinical parameters of BOT patients treated between 1998 and 2008 in 24 German centres were retrospectively investigated. Central pathology review and prospective follow-up were carried out. Patients <40 versus ≥40 years were analysed separately and then compared regarding clinico-pathological variables and prognosis. A total of 950 BOT patients with a median age of 49.1 (14.1-91.5) years were analysed [280 patients <40 years (29.5%), 670 patients ≥40 years (70.5%)]. Fertility-preserving surgery was carried out in 53.2% (149 of 280) of patients <40 years with preservation of the primarily affected ovary in 32 of these 149 cases (21.5%). Recurrence was significantly more frequent in patients <40 years (19.0% versus 10.1% 5-year recurrence rate, P < 0.001), usually in ovarian tissue, whereas disease-specific overall survival did not differ between the subgroups. In case of recurrent disease, malignant transformation was less frequent in younger than in older patients (12.0% versus 66.7%, P < 0.001), mostly presenting as invasive peritoneal carcinomatosis. Multivariate analysis for patients <40 years identified advanced International Federation of Gynecology and Obstetrics (FIGO) stage and fertility-sparing approach as independent prognostic factors negatively affecting progression-free survival (PFS) while, for patients ≥40 years, higher FIGO stage and incomplete staging was associated with impaired PFS. Despite favourable survival, young BOT patients with child-bearing potential are at higher risk for disease recurrence. However, relapses usually remain BOT in the preserved ovaries as opposed to older patients being at higher risk for malignant transformation in peritoneal or distant localisation. Therefore, fertility-sparing approach can be
Witters, Daan; Knez, Karel; Ceyssens, Frederik; Puers, Robert; Lammertyn, Jeroen
2013-06-07
Digital microfluidics is introduced as a novel platform with unique advantages for performing single-molecule detection. We demonstrate how superparamagnetic beads, used for capturing single protein molecules, can be printed with unprecedentedly high loading efficiency and single bead resolution on an electrowetting-on-dielectric-based digital microfluidic chip by micropatterning the Teflon-AF surface of the device. By transporting droplets containing suspended superparamagnetic beads over a hydrophilic-in-hydrophobic micropatterned Teflon-AF surface, single beads are trapped inside the hydrophilic microwells due to their selective wettability and tailored dimensions. Digital microfluidics presents the following advantages for printing and sealing magnetic beads for single-molecule detection: (i) droplets containing suspended beads can be transported back and forth over the array of hydrophilic microwells to obtain high loading efficiencies of microwells with single beads, (ii) the use of hydrophilic-in-hydrophobic patterns permits the use of a magnet to speed up the bead transfer process to the wells, while the receding droplet meniscus removes excess beads off the chip surface and thereby shortens the bead patterning time, and (iii) reagents can be transported over the printed beads multiple times, while capillary forces and a magnet hold the printed beads in place. High loading efficiencies (98% with a CV of 0.9%) of single beads in microwells were obtained by transporting droplets of suspended beads over the array 10 times in less than 1 min, which is much higher than previously reported methods (40-60%), while the total surface area needed for performing single-molecule detection can be decreased. The performance of the device was demonstrated by fluorescent detection of the presence of the biotinylated enzyme β-galactosidase on streptavidin-coated beads with a linear dynamic range of 4 orders of magnitude ranging from 10 aM to 90 fM.
SISGR: Room Temperature Single-Molecule Detection and Imaging by Stimulated Emission Microscopy
DOE Office of Scientific and Technical Information (OSTI.GOV)
Xie, Xiaoliang Sunney
Single-molecule spectroscopy has made considerable impact on many disciplines including chemistry, physics, and biology. To date, most single-molecule spectroscopy work is accomplished by detecting fluorescence. On the other hand, many naturally occurring chromophores, such as retinal, hemoglobin and cytochromes, do not have detectable fluorescence. There is an emerging need for single-molecule spectroscopy techniques that do not require fluorescence. In the last proposal period, we have successfully demonstrated stimulated emission microscopy, single molecule absorption, and stimulated Raman microscopy based on a high-frequency modulation transfer technique. These first-of-a- kind new spectroscopy/microscopy methods tremendously improved our ability to observe molecules that fluorescence weakly,more » even to the limit of single molecule detection for absorption measurement. All of these methods employ two laser beams: one (pump beam) excites a single molecule to a real or virtual excited state, and the other (probe beam) monitors the absorption/emission property of the single. We extract the intensity change of the probe beam with high sensitivity by implementing a high-frequency phase-sensitive detection scheme, which offers orders of magnitude improvement in detection sensitivity over direct absorption/emission measurement. However, single molecule detection based on fluorescence or absorption is fundamentally limited due to their broad spectral response. It is important to explore other avenues in single molecule detection and imaging which provides higher molecular specificity for studying a wide variety of heterogeneous chemical and biological systems. This proposal aimed to achieve single-molecule detection sensitivity with near resonance stimulated Raman scattering (SRS) microscopy. SRS microscopy was developed in our lab as a powerful technique for imaging heterogeneous samples based on their intrinsic vibrational contrasts, which provides much higher
An efficient method to detect periodic behavior in botnet traffic by analyzing control plane traffic
AsSadhan, Basil; Moura, José M.F.
2013-01-01
Botnets are large networks of bots (compromised machines) that are under the control of a small number of bot masters. They pose a significant threat to Internet’s communications and applications. A botnet relies on command and control (C2) communications channels traffic between its members for its attack execution. C2 traffic occurs prior to any attack; hence, the detection of botnet’s C2 traffic enables the detection of members of the botnet before any real harm happens. We analyze C2 traffic and find that it exhibits a periodic behavior. This is due to the pre-programmed behavior of bots that check for updates to download them every T seconds. We exploit this periodic behavior to detect C2 traffic. The detection involves evaluating the periodogram of the monitored traffic. Then applying Walker’s large sample test to the periodogram’s maximum ordinate in order to determine if it is due to a periodic component or not. If the periodogram of the monitored traffic contains a periodic component, then it is highly likely that it is due to a bot’s C2 traffic. The test looks only at aggregate control plane traffic behavior, which makes it more scalable than techniques that involve deep packet inspection (DPI) or tracking the communication flows of different hosts. We apply the test to two types of botnet, tinyP2P and IRC that are generated by SLINGbot. We verify the periodic behavior of their C2 traffic and compare it to the results we get on real traffic that is obtained from a secured enterprise network. We further study the characteristics of the test in the presence of injected HTTP background traffic and the effect of the duty cycle on the periodic behavior. PMID:25685512
Detecting Single-Nucleotide Substitutions Induced by Genome Editing.
Miyaoka, Yuichiro; Chan, Amanda H; Conklin, Bruce R
2016-08-01
The detection of genome editing is critical in evaluating genome-editing tools or conditions, but it is not an easy task to detect genome-editing events-especially single-nucleotide substitutions-without a surrogate marker. Here we introduce a procedure that significantly contributes to the advancement of genome-editing technologies. It uses droplet digital polymerase chain reaction (ddPCR) and allele-specific hydrolysis probes to detect single-nucleotide substitutions generated by genome editing (via homology-directed repair, or HDR). HDR events that introduce substitutions using donor DNA are generally infrequent, even with genome-editing tools, and the outcome is only one base pair difference in 3 billion base pairs of the human genome. This task is particularly difficult in induced pluripotent stem (iPS) cells, in which editing events can be very rare. Therefore, the technological advances described here have implications for therapeutic genome editing and experimental approaches to disease modeling with iPS cells. © 2016 Cold Spring Harbor Laboratory Press.
Koensgen, Dominique; Weiss, Martin; Assmann, Kathrin; Brucker, Sara Y; Wallwiener, Diethelm; Stope, Matthias B; Mustea, Alexander
2018-03-01
Borderline ovarian tumors (BOT) are malignant epithelial ovarian tumors with a very low incidence, therefore lacking sufficient clinical experience in diagnostics and treatment. This study characterized the histology, clinical features, diagnostics and therapy of BOT including patients treated at the Department of Gynecology and Obstetrics of the University Medicine Greifswald. In this retrospective, single-center study, patients with BOT treated between 1990 and 2010 were analyzed according to their histological and clinical reports. A total of 54 patients were enrolled. The median age was 54.6 (range=23-83) years. Distribution of histological subtypes was: serous in 31 patients (57.4 %) and mucinous in 23 patients (42.6%). All patients underwent surgery. Eight patients (14.8%) were treated according to actual therapy recommendations during the initial surgery. Eight patients (14.8%) received adjuvant chemotherapy contrary to treatment recommendations. In the case of 36 patients (66.7%), a frozen section was taken intraoperatively, which matched the definitive histological result in 88.9%. During average follow-up of 70.3 months (range=0-231 months), two patients (3.7%) developed tumor recurrence after 9 and 29 months, respectively, two patients (3.7%) died of causes other than BOT. Our study critically demonstrated that until a few years ago, BOTs were not usually treated according to international therapy recommendations. Nevertheless, the rate of tumor recurrence was very low. Copyright© 2018, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.
NASA Astrophysics Data System (ADS)
Payne, A.; Ambal, K.; Boehme, C.; Williams, C. C.
2015-05-01
A study of a force detected single-spin magnetic resonance measurement concept with atomic spatial resolution is presented. The method is based upon electrostatic force detection of spin-selection rule controlled single-electron tunneling between two electrically isolated paramagnetic states. Single-spin magnetic resonance detection is possible by measuring the force detected tunneling charge noise on and off spin resonance. Simulation results of this charge noise, based upon physical models of the tunneling and spin physics, are directly compared to measured atomic force microscopy system noise. The results show that the approach could provide single-spin measurement of electrically isolated qubit states with atomic spatial resolution at room temperature.
Kostev, Karel; Dippel, Franz-Werner
2012-12-01
To assess the predictors for the initiation of a basal supported oral therapy (BOT) in type 2 diabetic patients under real-life conditions in Germany. A historical cohort study based on representative German real life data (IMS(®) Disease Analyzer) was performed. The study included patients with type 2 diabetes who started an oral antidiabetic drug (OAD) treatment between 01/1995 and 12/2011. Patients with consecutive treatment data for at least 12 months before the initiation of an OAD treatment were eligible for the analysis. The time-dependent rate of patients starting an insulin therapy with a long-acting insulin was calculated by use of the Kaplan-Meier method. Multivariate Cox regression analyses were applied to identify associated factors. The study included 194,967 patients with type 2 diabetes mellitus being on OAD therapy. 24,964 patients were switched to BOT during the observational period. The probability of switching to insulin therapy was associated with three main predictors such as (1) poor metabolic control, (2) midlife age and (3) number and type of the OAD before insulinization. The variation of the HbA1c threshold to HbA1c≥7.5 leads to comparable outcomes with significant HR. The highest probability of initiating a basal supported oral therapy (BOT) under real life conditions was found for patients with poor metabolic control, midlife age and pre-treatment with specific OADs such as SU, GLI or AGI before initiation of insulin therapy. Copyright © 2012 Primary Care Diabetes Europe. Published by Elsevier Ltd. All rights reserved.
Ultrasound Visualization of Atypical Abscess Ultimately Containing Bot Fly Larva.
Bovino, Patrick; Cole, John; Scheatzle, Mark
2016-08-01
Because of the rise in community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA), presentations to the emergency department for the evaluation of cutaneous abscesses have risen dramatically over the past 2 decades. Soft tissue point of care ultrasound (POCUS) differentiates abscess from cellulitis, determines the size and shape, and characterizes the contents of the abscess. It has been shown to improve medical decision-making and therefore the emergency management of cutaneous abscesses over physical examination alone. We report a case of an unusual nonhealing abscess in an 18-year-old woman with a recent history of foreign travel where soft tissue POCUS identified motion within the abscess pocket. This changed the management of the case, leading to the diagnosis of bot fly myiasis. WHY SHOULD AN EMERGENCY PHYSICIAN BE AWARE OF THIS?: Clinicians should entertain a broader differential for an apparent abscess and consider liberal use of soft tissue POCUS in these cases. Copyright © 2016 Elsevier Inc. All rights reserved.
Detecting Peer-to-Peer Botnets in SCADA Systems
DOE Office of Scientific and Technical Information (OSTI.GOV)
Yang, Huan; Cheng, Liang; Chuah, Mooi Choo
Supervisory Control and Data Acquisition (SCADA) systems monitor and control critical infrastructure such as the smart grid. As SCADA systems become increasingly interconnected and adopt more and more cyber-enabled components, the risks of cyber attacks become a major concern. Due to their decentralized organization, peer-to-peer (P2P) botnets are resilient to many existing takedown measures and can be exploited as an effective way to launch cyber attacks on SCADA systems. However, little work has been done to detect P2P botnets in SCADA systems, which carry traffic flows with characteristics significantly different from the Internet. In this paper, we design a P2P-botnetmore » detection method for SCADA systems, leveraging built-in traffic monitoring capabilities of SCADA networking devices. The proposed method consists of two stages. In the first stage, we design a simple feature test to filter out non-P2P hosts, which significantly reduces the data volume for P2P-botnet identification. In the second stage, we jointly consider flow-based and connectivity-based features that effectively set apart bots from benign hosts. We propose to use unsupervised learning for P2P-botnet identification, which not only identifies known P2P botnets but also captures newly emerged ones. Our simulation results show that the proposed system achieves high detection rates with very few false positives. Furthermore, our evaluation shows that the proposed method can detect hosts running P2P SCADA applications that are infected by P2P bots.« less
NASA Astrophysics Data System (ADS)
Landry, Markita Patricia; Ando, Hiroki; Chen, Allen Y.; Cao, Jicong; Kottadiel, Vishal Isaac; Chio, Linda; Yang, Darwin; Dong, Juyao; Lu, Timothy K.; Strano, Michael S.
2017-05-01
A distinct advantage of nanosensor arrays is their ability to achieve ultralow detection limits in solution by proximity placement to an analyte. Here, we demonstrate label-free detection of individual proteins from Escherichia coli (bacteria) and Pichia pastoris (yeast) immobilized in a microfluidic chamber, measuring protein efflux from single organisms in real time. The array is fabricated using non-covalent conjugation of an aptamer-anchor polynucleotide sequence to near-infrared emissive single-walled carbon nanotubes, using a variable chemical spacer shown to optimize sensor response. Unlabelled RAP1 GTPase and HIV integrase proteins were selectively detected from various cell lines, via large near-infrared fluorescent turn-on responses. We show that the process of E. coli induction, protein synthesis and protein export is highly stochastic, yielding variability in protein secretion, with E. coli cells undergoing division under starved conditions producing 66% fewer secreted protein products than their non-dividing counterparts. We further demonstrate the detection of a unique protein product resulting from T7 bacteriophage infection of E. coli, illustrating that nanosensor arrays can enable real-time, single-cell analysis of a broad range of protein products from various cell types.
1990-03-28
observation, detection of the optical absorption of a single pentacene molecule in a p-terphenyl crystal, opens the door to new studies of single local ...produce appreciable quadratic Stark shifting. Such effects would greatly perturb the local field around the pentacene molecule, making detection of the...of the local surroundings of pentacene molecules with single injected charge carriers nearby may become an interesting field; however, for the
Ultrasensitive Laser Spectroscopy in Solids: Single-Molecule Detection
1989-10-25
spite of detection intensity constraints necessary to avoid power broadening, the optical absorption spectrum of single molecules of pentacene In p...molecule detection, or SMD) would provide a useful tool for the study of local host-absorber interactions where tihe absorbing ,ontor is essentially at...modulation techniques 7. 8 for the model system composed of pentacene substitutional impurities in p-terphenyl crystals at 1.5K. The pontacene molecules can
Microwave Detection With Single-Band Metamaterials For High Power Microwave Weapons
2017-12-01
DETECTION WITH SINGLE-BAND METAMATERIALS FOR HIGH POWER MICROWAVE WEAPONS by Edward V. Wulff December 2017 Thesis Advisor: Dragoslav Grbovic...3. REPORT TYPE AND DATES COVERED Master’s thesis 4. TITLE AND SUBTITLE MICROWAVE DETECTION WITH SINGLE-BAND METAMATERIALS FOR HIGH POWER...IRB number ____N/A____. 12a. DISTRIBUTION / AVAILABILITY STATEMENT Approved for public release. Distribution is unlimited. 12b. DISTRIBUTION
Rare Cell Detection by Single-Cell RNA Sequencing as Guided by Single-Molecule RNA FISH.
Torre, Eduardo; Dueck, Hannah; Shaffer, Sydney; Gospocic, Janko; Gupte, Rohit; Bonasio, Roberto; Kim, Junhyong; Murray, John; Raj, Arjun
2018-02-28
Although single-cell RNA sequencing can reliably detect large-scale transcriptional programs, it is unclear whether it accurately captures the behavior of individual genes, especially those that express only in rare cells. Here, we use single-molecule RNA fluorescence in situ hybridization as a gold standard to assess trade-offs in single-cell RNA-sequencing data for detecting rare cell expression variability. We quantified the gene expression distribution for 26 genes that range from ubiquitous to rarely expressed and found that the correspondence between estimates across platforms improved with both transcriptome coverage and increased number of cells analyzed. Further, by characterizing the trade-off between transcriptome coverage and number of cells analyzed, we show that when the number of genes required to answer a given biological question is small, then greater transcriptome coverage is more important than analyzing large numbers of cells. More generally, our report provides guidelines for selecting quality thresholds for single-cell RNA-sequencing experiments aimed at rare cell analyses. Copyright © 2018 Elsevier Inc. All rights reserved.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Ferrari, Simone; Kahl, Oliver; Kovalyuk, Vadim
We investigate single- and multi-photon detection regimes of superconducting nanowire detectors embedded in silicon nitride nanophotonic circuits. At near-infrared wavelengths, simultaneous detection of up to three photons is observed for 120 nm wide nanowires biased far from the critical current, while narrow nanowires below 100 nm provide efficient single photon detection. A theoretical model is proposed to determine the different detection regimes and to calculate the corresponding internal quantum efficiency. The predicted saturation of the internal quantum efficiency in the single photon regime agrees well with plateau behavior observed at high bias currents.
Direct detection of the optical field beyond single polarization mode.
Che, Di; Sun, Chuanbowen; Shieh, William
2018-02-05
Direct detection is traditionally regarded as a detection method that recovers only the optical intensity. Compared with coherent detection, it owns a natural advantage-the simplicity-but lacks a crucial capability of field recovery that enables not only the multi-dimensional modulation, but also the digital compensation of the fiber impairments linear with the optical field. Full-field detection is crucial to increase the capacity-distance product of optical transmission systems. A variety of methods have been investigated to directly detect the optical field of the single polarization mode, which normally sends a carrier traveling with the signal for self-coherent detection. The crux, however, is that any optical transmission medium supports at least two propagating modes (e.g. single mode fiber supports two polarization modes), and until now there is no direct detection that can recover the complete set of optical fields beyond one polarization, due to the well-known carrier fading issue after mode demultiplexing induced by the random mode coupling. To avoid the fading, direct detection receivers should recover the signal in an intensity space isomorphic to the optical field without loss of any degrees of freedom, and a bridge should be built between the field and its isomorphic space for the multi-mode field recovery. Based on this thinking, we propose, for the first time, the direct detection of dual polarization modes by a novel receiver concept, the Stokes-space field receiver (SSFR) and its extension, the generalized SSFR for multiple spatial modes. The idea is verified by a dual-polarization field recovery of a polarization-multiplexed complex signal over an 80-km single mode fiber transmission. SSFR can be applied to a much wider range of fields beyond optical communications such as coherent sensing and imaging, where simple field recovery without an extra local laser is desired for enhanced system performance.
Peck, K; Stryer, L; Glazer, A N; Mathies, R A
1989-01-01
A theory for single-molecule fluorescence detection is developed and then used to analyze data from subpicomolar solutions of B-phycoerythrin (PE). The distribution of detected counts is the convolution of a Poissonian continuous background with bursts arising from the passage of individual fluorophores through the focused laser beam. The autocorrelation function reveals single-molecule events and provides a criterion for optimizing experimental parameters. The transit time of fluorescent molecules through the 120-fl imaged volume was 800 microseconds. The optimal laser power (32 mW at 514.5 nm) gave an incident intensity of 1.8 x 10(23) photons.cm-2.s-1, corresponding to a mean time of 1.1 ns between absorptions. The mean incremental count rate was 1.5 per 100 microseconds for PE monomers and 3.0 for PE dimers above a background count rate of 1.0. The distribution of counts and the autocorrelation function for 200 fM monomer and 100 fM dimer demonstrate that single-molecule detection was achieved. At this concentration, the mean occupancy was 0.014 monomer molecules in the probed volume. A hard-wired version of this detection system was used to measure the concentration of PE down to 1 fM. This single-molecule counter is 3 orders of magnitude more sensitive than conventional fluorescence detection systems. PMID:2726766
Transforming the Detective Novel: Sex and the Single Sleuth.
ERIC Educational Resources Information Center
Marchino, Lois A.
The lesbian detective of current fiction may be single, but she is not alone; she has a lover. In varying degrees of realism and fantasy, sexuality is part of her character. While some writers in the field argue that romances involving the protagonists of feminist detective fiction distract from the plot, these critics themselves sometimes break…
Single photon detection in a waveguide-coupled Ge-on-Si lateral avalanche photodiode.
Martinez, Nicholas J D; Gehl, Michael; Derose, Christopher T; Starbuck, Andrew L; Pomerene, Andrew T; Lentine, Anthony L; Trotter, Douglas C; Davids, Paul S
2017-07-10
We examine gated-Geiger mode operation of an integrated waveguide-coupled Ge-on-Si lateral avalanche photodiode (APD) and demonstrate single photon detection at low dark count for this mode of operation. Our integrated waveguide-coupled APD is fabricated using a selective epitaxial Ge-on-Si growth process resulting in a separate absorption and charge multiplication (SACM) design compatible with our silicon photonics platform. Single photon detection efficiency and dark count rate is measured as a function of temperature in order to understand and optimize performance characteristics in this device. We report single photon detection of 5.27% at 1310 nm and a dark count rate of 534 kHz at 80 K for a Ge-on-Si single photon avalanche diode. Dark count rate is the lowest for a Ge-on-Si single photon detector in this range of temperatures while maintaining competitive detection efficiency. A jitter of 105 ps was measured for this device.
Detection of Single Molecules Illuminated by a Light-Emitting Diode
Gerhardt, Ilja; Mai, Lijian; Lamas-Linares, Antía; Kurtsiefer, Christian
2011-01-01
Optical detection and spectroscopy of single molecules has become an indispensable tool in biological imaging and sensing. Its success is based on fluorescence of organic dye molecules under carefully engineered laser illumination. In this paper we demonstrate optical detection of single molecules on a wide-field microscope with an illumination based on a commercially available, green light-emitting diode. The results are directly compared with laser illumination in the same experimental configuration. The setup and the limiting factors, such as light transfer to the sample, spectral filtering and the resulting signal-to-noise ratio are discussed. A theoretical and an experimental approach to estimate these parameters are presented. The results can be adapted to other single emitter and illumination schemes. PMID:22346610
Single Molecule Spectral Diffusion in a Solid Detected Via Fluorescence Spectroscopy
1991-10-15
other local fields) at the position of the molecule, the spectral jumps may occur because the class II pentacene molecules are coupled to an...and identify by block number) FIELD jGROUP SUB-GROUP_ Single molecule spectroscopy Precision detection Spectral diffusion, Pentacene in p-terphenyl 19...significant increases in detection sensitivity for single pentacene molecules in crystals of p-terphenyl at low temperatures. With the increased signal to
Rapid and efficient detection of single chromophore molecules in aqueous solution
NASA Astrophysics Data System (ADS)
Li, Li-Qiang; Davis, Lloyd M.
1995-06-01
The first experiments on the detection of single fluorescent molecules in a flowing stream of an aqueous solution with high total efficiency are reported. A capillary injection system for sample delivery causes all the dye molecules to pass in a diffusion-broadened stream within a fast-moving sheath flow, through the center of the tightly focused laser excitation beam. Single-molecule detection with a transit time of approximately 1 ms is accomplished with a high-quantum-efficiency single-photon avalanche diode and a low dead-time time-gating circuit for discrimination of Raman-scattered light from the solvent.
Single Molecule Detection in Living Biological Cells using Carbon Nanotube Optical Probes
NASA Astrophysics Data System (ADS)
Strano, Michael
2009-03-01
Nanoscale sensing elements offer promise for single molecule analyte detection in physically or biologically constrained environments. Molecular adsorption can be amplified via modulation of sharp singularities in the electronic density of states that arise from 1D quantum confinement [1]. Single-walled carbon nanotubes (SWNT), as single molecule optical sensors [2-3], offer unique advantages such as photostable near-infrared (n-IR) emission for prolonged detection through biological media, single-molecule sensitivity and, nearly orthogonal optical modes for signal transduction that can be used to identify distinct classes of analytes. Selective binding to the SWNT surface is difficult to engineer [4]. In this lecture, we will briefly review the immerging field of fluorescent diagnostics using band gap emission from SWNT. In recent work, we demonstrate that even a single pair of SWNT provides at least four optical modes that can be modulated to uniquely fingerprint chemical agents by the degree to which they alter either the emission band intensity or wavelength. We validate this identification method in vitro by demonstrating detection and identification of six genotoxic analytes, including chemotherapeutic drugs and reactive oxygen species (ROS), which are spectroscopically differentiated into four distinct classes. We also demonstrate single-molecule sensitivity in detecting hydrogen peroxide, one of the most common genotoxins and an important cellular signal. Finally, we employ our sensing and fingerprinting method of these analytes in real time within live 3T3 cells, demonstrating the first multiplexed optical detection from a nanoscale biosensor and the first label-free tool to optically discriminate between genotoxins. We will also discuss our recent efforts to fabricate biomedical sensors for real time detection of glucose and other important physiologically relevant analytes in-vivo. The response of embedded SWNT in a swellable hydrogel construct to
Kang, Dong-Ku; Ali, M. Monsur; Zhang, Kaixiang; Huang, Susan S.; Peterson, Ellena; Digman, Michelle A.; Gratton, Enrico; Zhao, Weian
2014-01-01
Blood stream infection or sepsis is a major health problem worldwide, with extremely high mortality, which is partly due to the inability to rapidly detect and identify bacteria in the early stages of infection. Here we present a new technology termed ‘Integrated Comprehensive Droplet Digital Detection’ (IC 3D) that can selectively detect bacteria directly from milliliters of diluted blood at single-cell sensitivity in a one-step, culture- and amplification-free process within 1.5–4 h. The IC 3D integrates real-time, DNAzyme-based sensors, droplet microencapsulation and a high-throughput 3D particle counter system. Using Escherichia coli as a target, we demonstrate that the IC 3D can provide absolute quantification of both stock and clinical isolates of E. coli in spiked blood within a broad range of extremely low concentration from 1 to 10,000 bacteria per ml with exceptional robustness and limit of detection in the single digit regime. PMID:25391809
Cucurbituril mediated single molecule detection and identification via recognition tunneling.
Xiao, Bohuai; Liang, Feng; Liu, Simin; Im, JongOne; Li, Yunchuan; Liu, Jing; Zhang, Bintian; Zhou, Jianghao; He, Jin; Chang, Shuai
2018-06-08
Recognition tunneling (RT) is an emerging technique for investigating single molecules in a tunnel junction. We have previously demonstrated its capability of single molecule detection and identification, as well as probing the dynamics of intermolecular bonding at the single molecule level. Here by introducing cucurbituril as a new class of recognition molecule, we demonstrate a powerful platform for electronically investigating the host-guest chemistry at single molecule level. In this report, we first investigated the single molecule electrical properties of cucurbituril in a tunnel junction. Then we studied two model guest molecules, aminoferrocene and amantadine, which were encapsulated by cucurbituril. Small differences in conductance and lifetime can be recognized between the host-guest complexes with the inclusion of different guest molecules. By using a machine learning algorithm to classify the RT signals in a hyper dimensional space, the accuracy of guest molecule recognition can be significantly improved, suggesting the possibility of using cucurbituril molecule for single molecule identification. This work enables a new class of recognition molecule for RT technique and opens the door for detecting a vast variety of small molecules by electrical measurements.
1989-08-18
CODES 18 SUBJECT TERMS (Continue on reverse if necessary and identify by block number) FIELD GROUP SUB-GROUP Single Molecule Detection Pentacene in p...and 10 additional pentacene molecules. This may be accomplished by- a combination of laser FM spectroscopy and either Stark or ultrasonic double...6099 408-927-2426 ABSTRACT: Single-absorber optical spectroscopy in solids is described for the case of finding a single pentacene molecule in a
Detecting single DNA molecule interactions with optical microcavities (Presentation Recording)
NASA Astrophysics Data System (ADS)
Vollmer, Frank
2015-09-01
Detecting molecules and their interactions lies at the heart of all biosensor devices, which have important applications in health, environmental monitoring and biomedicine. Achieving biosensing capability at the single molecule level is, moreover, a particularly important goal since single molecule biosensors would not only operate at the ultimate detection limit by resolving individual molecular interactions, but they could also monitor biomolecular properties which are otherwise obscured in ensemble measurements. For example, a single molecule biosensor could resolve the fleeting interaction kinetics between a molecule and its receptor, with immediate applications in clinical diagnostics. We have now developed a label-free biosensing platform that is capable of monitoring single DNA molecules and their interaction kinetics[1], hence achieving an unprecedented sensitivity in the optical domain, Figure 1. We resolve the specific contacts between complementary oligonucleotides, thereby detecting DNA strands with less than 2.4 kDa molecular weight. Furthermore we can discern strands with single nucleotide mismatches by monitoring their interaction kinetics. Our device utilizes small glass microspheres as optical transducers[1,2, 3], which are capable of increasing the number of interactions between a light beam and analyte molecules. A prism is used to couple the light beam into the microsphere. Ourr biosensing approach resolves the specific interaction kinetics between single DNA fragments. The optical transducer is assembled in a simple three-step protocol, and consists of a gold nanorod attached to a glass microsphere, where the surface of the nanorod is further modified with oligonucleotide receptors. The interaction kinetics of an oligonucleotide receptor with DNA fragments in the surrounding aqueous solution is monitored at the single molecule level[1]. The light remains confined inside the sphere where it is guided by total internal reflections along a
Direct detection of a single photon by humans
Tinsley, Jonathan N.; Molodtsov, Maxim I.; Prevedel, Robert; Wartmann, David; Espigulé-Pons, Jofre; Lauwers, Mattias; Vaziri, Alipasha
2016-01-01
Despite investigations for over 70 years, the absolute limits of human vision have remained unclear. Rod cells respond to individual photons, yet whether a single-photon incident on the eye can be perceived by a human subject has remained a fundamental open question. Here we report that humans can detect a single-photon incident on the cornea with a probability significantly above chance. This was achieved by implementing a combination of a psychophysics procedure with a quantum light source that can generate single-photon states of light. We further discover that the probability of reporting a single photon is modulated by the presence of an earlier photon, suggesting a priming process that temporarily enhances the effective gain of the visual system on the timescale of seconds. PMID:27434854
Detection of kinetic change points in piece-wise linear single molecule motion
NASA Astrophysics Data System (ADS)
Hill, Flynn R.; van Oijen, Antoine M.; Duderstadt, Karl E.
2018-03-01
Single-molecule approaches present a powerful way to obtain detailed kinetic information at the molecular level. However, the identification of small rate changes is often hindered by the considerable noise present in such single-molecule kinetic data. We present a general method to detect such kinetic change points in trajectories of motion of processive single molecules having Gaussian noise, with a minimum number of parameters and without the need of an assumed kinetic model beyond piece-wise linearity of motion. Kinetic change points are detected using a likelihood ratio test in which the probability of no change is compared to the probability of a change occurring, given the experimental noise. A predetermined confidence interval minimizes the occurrence of false detections. Applying the method recursively to all sub-regions of a single molecule trajectory ensures that all kinetic change points are located. The algorithm presented allows rigorous and quantitative determination of kinetic change points in noisy single molecule observations without the need for filtering or binning, which reduce temporal resolution and obscure dynamics. The statistical framework for the approach and implementation details are discussed. The detection power of the algorithm is assessed using simulations with both single kinetic changes and multiple kinetic changes that typically arise in observations of single-molecule DNA-replication reactions. Implementations of the algorithm are provided in ImageJ plugin format written in Java and in the Julia language for numeric computing, with accompanying Jupyter Notebooks to allow reproduction of the analysis presented here.
Single molecule targeted sequencing for cancer gene mutation detection.
Gao, Yan; Deng, Liwei; Yan, Qin; Gao, Yongqian; Wu, Zengding; Cai, Jinsen; Ji, Daorui; Li, Gailing; Wu, Ping; Jin, Huan; Zhao, Luyang; Liu, Song; Ge, Liangjin; Deem, Michael W; He, Jiankui
2016-05-19
With the rapid decline in cost of sequencing, it is now affordable to examine multiple genes in a single disease-targeted clinical test using next generation sequencing. Current targeted sequencing methods require a separate step of targeted capture enrichment during sample preparation before sequencing. Although there are fast sample preparation methods available in market, the library preparation process is still relatively complicated for physicians to use routinely. Here, we introduced an amplification-free Single Molecule Targeted Sequencing (SMTS) technology, which combined targeted capture and sequencing in one step. We demonstrated that this technology can detect low-frequency mutations using artificially synthesized DNA sample. SMTS has several potential advantages, including simple sample preparation thus no biases and errors are introduced by PCR reaction. SMTS has the potential to be an easy and quick sequencing technology for clinical diagnosis such as cancer gene mutation detection, infectious disease detection, inherited condition screening and noninvasive prenatal diagnosis.
Single-protein detection in crowded molecular environments in cryo-EM images
Rickgauer, J Peter; Grigorieff, Nikolaus; Denk, Winfried
2017-01-01
We present an approach to study macromolecular assemblies by detecting component proteins’ characteristic high-resolution projection patterns, calculated from their known 3D structures, in single electron cryo-micrographs. Our method detects single apoferritin molecules in vitreous ice with high specificity and determines their orientation and location precisely. Simulations show that high spatial-frequency information and—in the presence of protein background—a whitening filter are essential for optimal detection, in particular for images taken far from focus. Experimentally, we could detect small viral RNA polymerase molecules, distributed randomly among binding locations, inside rotavirus particles. Based on the currently attainable image quality, we estimate a threshold for detection that is 150 kDa in ice and 300 kDa in 100 nm thick samples of dense biological material. DOI: http://dx.doi.org/10.7554/eLife.25648.001 PMID:28467302
DOE Office of Scientific and Technical Information (OSTI.GOV)
DeMarini, D.M.; Brooks, H.G.
1992-01-01
Twenty-eight chlorinated organic compounds were evaluated for their ability to induce DNA damage using the Microscreen prophage-induction assay in Escherichia coli. Comparison of the performance characteristics of the prophage-induction and Salmonella assays to rodent carcinogenicity assays showed that the prophage-induction assay had a somewhat higher specificity than did the Salmonella assay (70% vs. 50%); sensitivity, concordance, and positive and negative predictivity were similar for the two microbial assays. The Microscreen prophage-induction assay failed to detect eight carcinogens, perhaps due to toxicity or other unknown factors; five of these eight carcinogens were detected by the Salmonella assay. However, the prophage-induction assaymore » did detect six carcinogens that were not detected by the Salmonella assay, and five of these were single-species, single-site carcinogens, mostly mouse liver carcinogens. Some of these carcinogens, such as the chloroethanes, produce free radicals, which may be the basis for their carcinogenicity and ability to induce prophage. The prophage-induction (or other SOS) assay may be useful in identifying some genotoxic chlorinated carcinogens that induce DNA damage that do not revert the standard Salmonella tester strains.« less
Detection of single nano-defects in photonic crystals between crossed polarizers.
Grepstad, Jon Olav; Kaspar, Peter; Johansen, Ib-Rune; Solgaard, Olav; Sudbø, Aasmund
2013-12-16
We investigate, by simulations and experiments, the light scattering of small particles trapped in photonic crystal membranes supporting guided resonance modes. Our results show that, due to amplified Rayleigh small particle scattering, such membranes can be utilized to make a sensor that can detect single nano-particles. We have designed a biomolecule sensor that uses cross-polarized excitation and detection for increased sensitivity. Estimated using Rayleigh scattering theory and simulation results, the current fabricated sensor has a detection limit of 26 nm, corresponding to the size of a single virus. The sensor can potentially be made both cheap and compact, to facilitate use at point-of-care.
Direct single-molecule dynamic detection of chemical reactions.
Guan, Jianxin; Jia, Chuancheng; Li, Yanwei; Liu, Zitong; Wang, Jinying; Yang, Zhongyue; Gu, Chunhui; Su, Dingkai; Houk, Kendall N; Zhang, Deqing; Guo, Xuefeng
2018-02-01
Single-molecule detection can reveal time trajectories and reaction pathways of individual intermediates/transition states in chemical reactions and biological processes, which is of fundamental importance to elucidate their intrinsic mechanisms. We present a reliable, label-free single-molecule approach that allows us to directly explore the dynamic process of basic chemical reactions at the single-event level by using stable graphene-molecule single-molecule junctions. These junctions are constructed by covalently connecting a single molecule with a 9-fluorenone center to nanogapped graphene electrodes. For the first time, real-time single-molecule electrical measurements unambiguously show reproducible large-amplitude two-level fluctuations that are highly dependent on solvent environments in a nucleophilic addition reaction of hydroxylamine to a carbonyl group. Both theoretical simulations and ensemble experiments prove that this observation originates from the reversible transition between the reactant and a new intermediate state within a time scale of a few microseconds. These investigations open up a new route that is able to be immediately applied to probe fast single-molecule physics or biophysics with high time resolution, making an important contribution to broad fields beyond reaction chemistry.
Direct single-molecule dynamic detection of chemical reactions
Guan, Jianxin; Jia, Chuancheng; Li, Yanwei; Liu, Zitong; Wang, Jinying; Yang, Zhongyue; Gu, Chunhui; Su, Dingkai; Houk, Kendall N.; Zhang, Deqing; Guo, Xuefeng
2018-01-01
Single-molecule detection can reveal time trajectories and reaction pathways of individual intermediates/transition states in chemical reactions and biological processes, which is of fundamental importance to elucidate their intrinsic mechanisms. We present a reliable, label-free single-molecule approach that allows us to directly explore the dynamic process of basic chemical reactions at the single-event level by using stable graphene-molecule single-molecule junctions. These junctions are constructed by covalently connecting a single molecule with a 9-fluorenone center to nanogapped graphene electrodes. For the first time, real-time single-molecule electrical measurements unambiguously show reproducible large-amplitude two-level fluctuations that are highly dependent on solvent environments in a nucleophilic addition reaction of hydroxylamine to a carbonyl group. Both theoretical simulations and ensemble experiments prove that this observation originates from the reversible transition between the reactant and a new intermediate state within a time scale of a few microseconds. These investigations open up a new route that is able to be immediately applied to probe fast single-molecule physics or biophysics with high time resolution, making an important contribution to broad fields beyond reaction chemistry. PMID:29487914
Single bead detection with an NMR microcapillary probe.
Nakashima, Yoshihiro; Boss, Michael; Russek, Stephen E; Moreland, John
2012-11-01
We have developed a nuclear magnetic resonance (NMR) microcapillary probe for the detection of single magnetic microbeads. The geometry of the probe has been optimized so that the signal from the background water has a similar magnitude compared to the signal from the dephased water nearby a single magnetic bead within the probe detector coil. In addition, the RF field of the coil must be uniform within the effective range of the magnetic bead. Three different RF probes were tested in a 7 T (300 MHz) pulsed NMR spectrometer with sample volumes ranging from 5 nL down to 1 nL. The 1 nL probe had a single-shot signal-to-noise ratio (SNR) for pure water of 27 and a volume resolution that exhibits a 600-fold improvement over a conventional (5 mm tube) NMR probe with a sample volume of 18 μL. This allowed for the detection of a 1 μm magnetite/polystyrene bead (m=2×10(-14)Am(2)) with an estimated experimental SNR of 30. Simulations of the NMR spectra for the different coil geometries and positions of the bead within the coil were developed that include the B(0) shift near a single bead, the inhomogeneity of the coils, the local coil sensitivity, the skin effect of the coil conductor, and quantitated estimates of the proximity effect between coil windings. Published by Elsevier Inc.
Li, Chen-Chen; Zhang, Yan; Tang, Bo; Zhang, Chun-Yang
2018-06-05
We combine single-molecule detection with magnetic separation for simultaneous measurement of human 8-oxoG DNA glycosylase 1 (hOGG1) and uracil DNA glycosylase (UDG) based on excision repair-initiated endonuclease IV (Endo IV)-assisted signal amplification. This method can sensitively detect multiple DNA glycosylases, and it can be further applied for the simultaneous measurement of enzyme kinetic parameters and screening of both hOGG1 and UDG inhibitors.
Single-Shot Quantum Nondemolition Detection of Individual Itinerant Microwave Photons
NASA Astrophysics Data System (ADS)
Besse, Jean-Claude; Gasparinetti, Simone; Collodo, Michele C.; Walter, Theo; Kurpiers, Philipp; Pechal, Marek; Eichler, Christopher; Wallraff, Andreas
2018-04-01
Single-photon detection is an essential component in many experiments in quantum optics, but it remains challenging in the microwave domain. We realize a quantum nondemolition detector for propagating microwave photons and characterize its performance using a single-photon source. To this aim, we implement a cavity-assisted conditional phase gate between the incoming photon and a superconducting artificial atom. By reading out the state of this atom in a single shot, we reach an external (internal) photon-detection fidelity of 50% (71%), limited by transmission efficiency between the source and the detector (75%) and the coherence properties of the qubit. By characterizing the coherence and average number of photons in the field reflected off the detector, we demonstrate its quantum nondemolition nature. We envisage applications in generating heralded remote entanglement between qubits and for realizing logic gates between propagating microwave photons.
Anti-dynamic-crosstalk method for single photon LIDAR detection
NASA Astrophysics Data System (ADS)
Zhang, Fan; Liu, Qiang; Gong, Mali; Fu, Xing
2017-11-01
With increasing number of vehicles equipped with light detection and ranging (LIDAR), crosstalk is identified as a critical and urgent issue in the range detection for active collision avoidance. Chaotic pulse position modulation (CPPM) applied in the transmitting pulse train has been shown to prevent crosstalk as well as range ambiguity. However, static and unified strategy on discrimination threshold and the number of accumulated pulse is not valid against crosstalk with varying number of sources and varying intensity of each source. This paper presents an adaptive algorithm to distinguish the target echo from crosstalk with dynamic and unknown level of intensity in the context of intelligent vehicles. New strategy is given based on receiver operating characteristics (ROC) curves that consider the detection requirements of the probability of detection and false alarm for the scenario with varying crosstalk. In the adaptive algorithm, the detected results are compared by the new strategy with both the number of accumulated pulses and the threshold being raised step by step, so that the target echo can be exactly identified from crosstalk with the dynamic and unknown level of intensity. The validity of the algorithm has been verified through the experiments with a single photon detector and the time correlated single photo counting (TCSPC) technique, demonstrating a marked drop in required shots for identifying the target compared with static and unified strategy
Distributed Detection with Collisions in a Random, Single-Hop Wireless Sensor Network
2013-05-26
public release; distribution is unlimited. Distributed detection with collisions in a random, single-hop wireless sensor network The views, opinions...1274 2 ABSTRACT Distributed detection with collisions in a random, single-hop wireless sensor network Report Title We consider the problem of... WIRELESS SENSOR NETWORK Gene T. Whipps?† Emre Ertin† Randolph L. Moses† ?U.S. Army Research Laboratory, Adelphi, MD 20783 †The Ohio State University
Detecting single viruses and nanoparticles using whispering gallery microlasers.
He, Lina; Ozdemir, Sahin Kaya; Zhu, Jiangang; Kim, Woosung; Yang, Lan
2011-06-26
There is a strong demand for portable systems that can detect and characterize individual pathogens and other nanoscale objects without the use of labels, for applications in human health, homeland security, environmental monitoring and diagnostics. However, most nanoscale objects of interest have low polarizabilities due to their small size and low refractive index contrast with the surrounding medium. This leads to weak light-matter interactions, and thus makes the label-free detection of single nanoparticles very difficult. Micro- and nano-photonic devices have emerged as highly sensitive platforms for such applications, because the combination of high quality factor Q and small mode volume V leads to significantly enhanced light-matter interactions. For example, whispering gallery mode microresonators have been used to detect and characterize single influenza virions and polystyrene nanoparticles with a radius of 30 nm (ref. 12) by measuring in the transmission spectrum either the resonance shift or mode splitting induced by the nanoscale objects. Increasing Q leads to a narrower resonance linewidth, which makes it possible to resolve smaller changes in the transmission spectrum, and thus leads to improved performance. Here, we report a whispering gallery mode microlaser-based real-time and label-free detection method that can detect individual 15-nm-radius polystyrene nanoparticles, 10-nm gold nanoparticles and influenza A virions in air, and 30 nm polystyrene nanoparticles in water. Our approach relies on measuring changes in the beat note that is produced when an ultra-narrow emission line from a whispering gallery mode microlaser is split into two modes by a nanoscale object, and these two modes then interfere. The ultimate detection limit is set by the laser linewidth, which can be made much narrower than the resonance linewidth of any passive resonator. This means that microlaser sensors have the potential to detect objects that are too small to be
JacksonBot - Design, Simulation and Optimal Control of an Action Painting Robot
NASA Astrophysics Data System (ADS)
Raschke, Michael; Mombaur, Katja; Schubert, Alexander
We present the robotics platform JacksonBot which is capable to produce paintings inspired by the Action Painting style of Jackson Pollock. A dynamically moving robot arm splashes color from a container at the end effector on the canvas. The paintings produced by this platform rely on a combination of the algorithmic generation of robot arm motions with random effects of the splashing color. The robot can be considered as a complex and powerful tool to generate art works programmed by a user. Desired end effector motions can be prescribed either by mathematical functions, by point sequences or by data glove motions. We have evaluated the effect of different shapes of input motions on the resulting painting. In order to compute the robot joint trajectories necessary to move along a desired end effector path, we use an optimal control based approach to solve the inverse kinematics problem.
32-channel single photon counting module for ultrasensitive detection of DNA sequences
NASA Astrophysics Data System (ADS)
Gudkov, Georgiy; Dhulla, Vinit; Borodin, Anatoly; Gavrilov, Dmitri; Stepukhovich, Andrey; Tsupryk, Andrey; Gorbovitski, Boris; Gorfinkel, Vera
2006-10-01
We continue our work on the design and implementation of multi-channel single photon detection systems for highly sensitive detection of ultra-weak fluorescence signals, for high-performance, multi-lane DNA sequencing instruments. A fiberized, 32-channel single photon detection (SPD) module based on single photon avalanche diode (SPAD), model C30902S-DTC, from Perkin Elmer Optoelectronics (PKI) has been designed and implemented. Unavailability of high performance, large area SPAD arrays and our desire to design high performance photon counting systems drives us to use individual diodes. Slight modifications in our quenching circuit has doubled the linear range of our system from 1MHz to 2MHz, which is the upper limit for these devices and the maximum saturation count rate has increased to 14 MHz. The detector module comprises of a single board computer PC-104 that enables data visualization, recording, processing, and transfer. Very low dark count (300-1000 counts/s), robust, efficient, simple data collection and processing, ease of connectivity to any other application demanding similar requirements and similar performance results to the best commercially available single photon counting module (SPCM from PKI) are some of the features of this system.
Revenue Risk Modelling and Assessment on BOT Highway Project
NASA Astrophysics Data System (ADS)
Novianti, T.; Setyawan, H. Y.
2018-01-01
The infrastructure project which is considered as a public-private partnership approach under BOT (Build-Operate-Transfer) arrangement, such as a highway, is risky. Therefore, assessment on risk factors is essential as the project have a concession period and is influenced by macroeconomic factors and consensus period. In this study, pre-construction risks of a highway were examined by using a Delphi method to create a space for offline expert discussions; a fault tree analysis to map intuition of experts and to create a model from the underlying risk events; a fuzzy logic to interpret the linguistic data of risk models. The loss of revenue for risk tariff, traffic volume, force majeure, and income were then measured. The results showed that the loss of revenue caused by the risk tariff was 10.5% of the normal total revenue. The loss of revenue caused by the risk of traffic volume was 21.0% of total revenue. The loss of revenue caused by the force majeure was 12.2% of the normal income. The loss of income caused by the non-revenue events was 6.9% of the normal revenue. It was also found that the volume of traffic was the major risk of a highway project because it related to customer preferences.
In situ single cell detection via microfluidic magnetic bead assay
KC, Pawan; Zhang, Ge; Zhe, Jiang
2017-01-01
We present a single cell detection device based on magnetic bead assay and micro Coulter counters. This device consists of two successive micro Coulter counters, coupled with a high gradient magnetic field generated by an external magnet. The device can identify single cells in terms of the transit time difference of the cell through the two micro Coulter counters. Target cells are conjugated with magnetic beads via specific antibody and antigen binding. A target cell traveling through the two Coulter counters interacts with the magnetic field, and have a longer transit time at the 1st counter than that at the 2nd counter. In comparison, a non-target cell has no interaction with the magnetic field, and hence has nearly the same transit times through the two counters. Each cell passing through the two counters generates two consecutive voltage pulses one after the other; the pulse widths and magnitudes indicating the cell’s transit times through the counters and the cell’s size respectively. Thus, by measuring the pulse widths (transit times) of each cell through the two counters, each single target cell can be differentiated from non-target cells even if they have similar sizes. We experimentally proved that the target human umbilical vein endothelial cells (HUVECs) and non-target rat adipose-derived stem cells (rASCs) have significant different transit time distribution, from which we can determine the recognition regions for both cell groups quantitatively. We further demonstrated that within a mixed cell population of rASCs and HUVECs, HUVECs can be detected in situ and the measured HUVECs ratios agree well with the pre-set ratios. With the simple device structure and easy sample preparation, this method is expected to enable single cell detection in a continuous flow and can be applied to facilitate general cell detection applications such as stem cell identification and enumeration. PMID:28222140
In situ single cell detection via microfluidic magnetic bead assay.
Liu, Fan; Kc, Pawan; Zhang, Ge; Zhe, Jiang
2017-01-01
We present a single cell detection device based on magnetic bead assay and micro Coulter counters. This device consists of two successive micro Coulter counters, coupled with a high gradient magnetic field generated by an external magnet. The device can identify single cells in terms of the transit time difference of the cell through the two micro Coulter counters. Target cells are conjugated with magnetic beads via specific antibody and antigen binding. A target cell traveling through the two Coulter counters interacts with the magnetic field, and have a longer transit time at the 1st counter than that at the 2nd counter. In comparison, a non-target cell has no interaction with the magnetic field, and hence has nearly the same transit times through the two counters. Each cell passing through the two counters generates two consecutive voltage pulses one after the other; the pulse widths and magnitudes indicating the cell's transit times through the counters and the cell's size respectively. Thus, by measuring the pulse widths (transit times) of each cell through the two counters, each single target cell can be differentiated from non-target cells even if they have similar sizes. We experimentally proved that the target human umbilical vein endothelial cells (HUVECs) and non-target rat adipose-derived stem cells (rASCs) have significant different transit time distribution, from which we can determine the recognition regions for both cell groups quantitatively. We further demonstrated that within a mixed cell population of rASCs and HUVECs, HUVECs can be detected in situ and the measured HUVECs ratios agree well with the pre-set ratios. With the simple device structure and easy sample preparation, this method is expected to enable single cell detection in a continuous flow and can be applied to facilitate general cell detection applications such as stem cell identification and enumeration.
Single Nanochannel-Aptamer-Based Biosensor for Ultrasensitive and Selective Cocaine Detection.
Wang, Jian; Hou, Jue; Zhang, Huacheng; Tian, Ye; Jiang, Lei
2018-01-17
Ultrasensitive and selective detection of molecules at nano or sub-nanomolar level is very important for many areas such as early diagnosis and drug testing. Herein, we report a high-sensitive cocaine sensor based on a single nanochannel coupled with DNA aptamers. The single nanochannel-aptamer-based biosensor can recognize cocaine molecules with an excellent sensitivity and good selectivity. A linear relationship between target cocaine concentration and output ionic current is obtained in a wide concentration range of cocaine from 1 nM to 10 μM. The cocaine sensor also shows a detection limit down to 1 nM. This study provides a new avenue to develop new nanochannel-aptamer-based biosensors for rapid and ultratrace detection of a variety of illicit drugs.
Communication: atomic force detection of single-molecule nonlinear optical vibrational spectroscopy.
Saurabh, Prasoon; Mukamel, Shaul
2014-04-28
Atomic Force Microscopy (AFM) allows for a highly sensitive detection of spectroscopic signals. This has been first demonstrated for NMR of a single molecule and recently extended to stimulated Raman in the optical regime. We theoretically investigate the use of optical forces to detect time and frequency domain nonlinear optical signals. We show that, with proper phase matching, the AFM-detected signals closely resemble coherent heterodyne-detected signals. Applications are made to AFM-detected and heterodyne-detected vibrational resonances in Coherent Anti-Stokes Raman Spectroscopy (χ((3))) and sum or difference frequency generation (χ((2))).
Osche, G R
2000-08-20
Single- and multiple-pulse detection statistics are presented for aperture-averaged direct detection optical receivers operating against partially developed speckle fields. A partially developed speckle field arises when the probability density function of the received intensity does not follow negative exponential statistics. The case of interest here is the target surface that exhibits diffuse as well as specular components in the scattered radiation. An approximate expression is derived for the integrated intensity at the aperture, which leads to single- and multiple-pulse discrete probability density functions for the case of a Poisson signal in Poisson noise with an additive coherent component. In the absence of noise, the single-pulse discrete density function is shown to reduce to a generalized negative binomial distribution. The radar concept of integration loss is discussed in the context of direct detection optical systems where it is shown that, given an appropriate set of system parameters, multiple-pulse processing can be more efficient than single-pulse processing over a finite range of the integration parameter n.
NASA Astrophysics Data System (ADS)
Bossu, R.; Etivant, C.; Roussel, F.; Mazet-Roux, G.; Steed, R.
2014-12-01
Smartphone applications have swiftly become one of the most popular tools for rapid reception of earthquake information for the public. Wherever someone's own location is, they can be automatically informed when an earthquake has struck just by setting a magnitude threshold and an area of interest. No need to browse the internet: the information reaches you automatically and instantaneously! One question remains: are the provided earthquake notifications always relevant for the public? A while after damaging earthquakes many eyewitnesses scrap the application they installed just after the mainshock. Why? Because either the magnitude threshold is set too high and many felt earthquakes are missed, or it is set too low and the majority of the notifications are related to unfelt earthquakes thereby only increasing anxiety among the population at each new update. Felt and damaging earthquakes are the ones of societal importance even when of small magnitude. LastQuake app and Twitter feed (QuakeBot) focuses on these earthquakes that matter for the public by collating different information threads covering tsunamigenic, damaging and felt earthquakes. Non-seismic detections and macroseismic questionnaires collected online are combined to identify felt earthquakes regardless their magnitude. Non seismic detections include Twitter earthquake detections, developed by the USGS, where the number of tweets containing the keyword "earthquake" is monitored in real time and flashsourcing, developed by the EMSC, which detect traffic surges on its rapid earthquake information website caused by the natural convergence of eyewitnesses who rush to the Internet to investigate the cause of the shaking that they have just felt. We will present the identification process of the felt earthquakes, the smartphone application and the 27 automatically generated tweets and how, by providing better public services, we collect more data from citizens.
An integrated optics microfluidic device for detecting single DNA molecules.
Krogmeier, Jeffrey R; Schaefer, Ian; Seward, George; Yantz, Gregory R; Larson, Jonathan W
2007-12-01
A fluorescence-based integrated optics microfluidic device is presented, capable of detecting single DNA molecules in a high throughput and reproducible manner. The device integrates microfluidics for DNA stretching with two optical elements for single molecule detection (SMD): a plano-aspheric refractive lens for fluorescence excitation (illuminator) and a solid parabolic reflective mirror for fluorescence collection (collector). Although miniaturized in size, both optical components were produced and assembled onto the microfluidic device by readily manufacturable fabrication techniques. The optical resolution of the device is determined by the small and relatively low numerical aperture (NA) illuminator lens (0.10 effective NA, 4.0 mm diameter) that delivers excitation light to a diffraction limited 2.0 microm diameter spot at full width half maximum within the microfluidic channel. The collector (0.82 annular NA, 15 mm diameter) reflects the fluorescence over a large collection angle, representing 71% of a hemisphere, toward a single photon counting module in an infinity-corrected scheme. As a proof-of-principle experiment for this simple integrated device, individual intercalated lambda-phage DNA molecules (48.5 kb) were stretched in a mixed elongational-shear microflow, detected, and sized with a fluorescence signal to noise ratio of 9.9 +/-1.0. We have demonstrated that SMD does not require traditional high numerical aperture objective lenses and sub-micron positioning systems conventionally used in many applications. Rather, standard manufacturing processes can be combined in a novel way that promises greater accessibility and affordability for microfluidic-based single molecule applications.
The whispering gallery mode biosensor: label-free detection from virus to single protein
NASA Astrophysics Data System (ADS)
Holler, S.; Dantham, V. R.; Keng, D.; Kolchenko, V.; Arnold, S.; Mulroe, Brigid; Paspaley-Grbavac, M.
2014-08-01
The whispering gallery mode (WGM) biosensor is a micro-optical platform capable of sensitive label-free detection of biological particles. Described by the reactive sensing principle (RSP), this analytic formulation quantifies the response of the system to the adsorption of bioparticles. Guided by the RSP, the WGM biosensor enabling from detection of virus (e.g., Human Papillomavirus, HPV) to the ultimate goal of single protein detection. The latter was derived from insights into the RSP, which resulted in the development of a hybrid plasmonic WGM biosensor, which has recently demonstrated detection of individual protein cancer markers. Enhancements from bound gold nanoparticles provide the sensitivity to detect single protein molecules (66 kDa) with good signal-to-noise (S/N > 10), and project that detection of proteins as small as 5 kDa.
Theory of single-photon detectors employing smart strategies of detection
NASA Astrophysics Data System (ADS)
Silva, João Batista Rosa; Ramos, Rubens Viana
2005-11-01
Single-photon detectors have become more important with the advent of set-ups for optical communication using single-photon pulses, mainly quantum key distribution. The performance of quantum key distribution systems depends strongly on the performance of single-photon detectors. In this paper, aiming to overcome the afterpulsing that limits strongly the maximal transmission rate of quantum key distribution systems, three smart strategies for single-photon detection are discussed using analytical and numerical procedures. The three strategies are: hold-off time conditioned to avalanche presence, termed the Norwegian strategy, using one avalanche photodiode, using two raffled avalanche photodiodes and using two switched avalanche photodiodes. Finally we give examples using these strategies in a quantum key distribution set-up.
Single-Molecule Tracking and Its Application in Biomolecular Binding Detection
Liu, Cong; Liu, Yen-Liang; Perillo, Evan P.; Dunn, Andrew K.; Yeh, Hsin-Chih
2016-01-01
In the past two decades significant advances have been made in single-molecule detection, which enables the direct observation of single biomolecules at work in real time and under physiological conditions. In particular, the development of single-molecule tracking (SMT) microscopy allows us to monitor the motion paths of individual biomolecules in living systems, unveiling the localization dynamics and transport modalities of the biomolecules that support the development of life. Beyond the capabilities of traditional camera-based tracking techniques, state-of-the-art SMT microscopies developed in recent years can record fluorescence lifetime while tracking a single molecule in the 3D space. This multiparameter detection capability can open the door to a wide range of investigations at the cellular or tissue level, including identification of molecular interaction hotspots and characterization of association/dissociation kinetics between molecules. In this review, we discuss various SMT techniques developed to date, with an emphasis on our recent development of the next generation 3D tracking system that not only achieves ultrahigh spatiotemporal resolution but also provides sufficient working depth suitable for live animal imaging. We also discuss the challenges that current SMT techniques are facing and the potential strategies to tackle those challenges. PMID:27660404
Single-Molecule Tracking and Its Application in Biomolecular Binding Detection.
Liu, Cong; Liu, Yen-Liang; Perillo, Evan P; Dunn, Andrew K; Yeh, Hsin-Chih
2016-01-01
In the past two decades significant advances have been made in single-molecule detection, which enables the direct observation of single biomolecules at work in real time and under physiological conditions. In particular, the development of single-molecule tracking (SMT) microscopy allows us to monitor the motion paths of individual biomolecules in living systems, unveiling the localization dynamics and transport modalities of the biomolecules that support the development of life. Beyond the capabilities of traditional camera-based tracking techniques, state-of-the-art SMT microscopies developed in recent years can record fluorescence lifetime while tracking a single molecule in the 3D space. This multiparameter detection capability can open the door to a wide range of investigations at the cellular or tissue level, including identification of molecular interaction hotspots and characterization of association/dissociation kinetics between molecules. In this review, we discuss various SMT techniques developed to date, with an emphasis on our recent development of the next generation 3D tracking system that not only achieves ultrahigh spatiotemporal resolution but also provides sufficient working depth suitable for live animal imaging. We also discuss the challenges that current SMT techniques are facing and the potential strategies to tackle those challenges.
Wavefront detection method of a single-sensor based adaptive optics system.
Wang, Chongchong; Hu, Lifa; Xu, Huanyu; Wang, Yukun; Li, Dayu; Wang, Shaoxin; Mu, Quanquan; Yang, Chengliang; Cao, Zhaoliang; Lu, Xinghai; Xuan, Li
2015-08-10
In adaptive optics system (AOS) for optical telescopes, the reported wavefront sensing strategy consists of two parts: a specific sensor for tip-tilt (TT) detection and another wavefront sensor for other distortions detection. Thus, a part of incident light has to be used for TT detection, which decreases the light energy used by wavefront sensor and eventually reduces the precision of wavefront correction. In this paper, a single Shack-Hartmann wavefront sensor based wavefront measurement method is presented for both large amplitude TT and other distortions' measurement. Experiments were performed for testing the presented wavefront method and validating the wavefront detection and correction ability of the single-sensor based AOS. With adaptive correction, the root-mean-square of residual TT was less than 0.2 λ, and a clear image was obtained in the lab. Equipped on a 1.23-meter optical telescope, the binary stars with angle distance of 0.6″ were clearly resolved using the AOS. This wavefront measurement method removes the separate TT sensor, which not only simplifies the AOS but also saves light energy for subsequent wavefront sensing and imaging, and eventually improves the detection and imaging capability of the AOS.
Wang, Yonghong; Wang, Ping; Wang, Yiqiang; He, Xiaoxiao; Wang, Kemin
2015-08-15
In this work, a simple and sensitive electrochemical strategy for arsenite detection based on the ability of arsenite bound to single-strand DNA (ssDNA) and the signal transduction of single wall carbon nanotubes (SWCNTs) is developed. To realize this purpose, the ssDNA/SWCNTs complexes were formed at first by making ssDNA wrapped around SWCNTs via π-stacking. In the presence of arsenite, the arsenite could strongly bind with the G/T bases of ssDNA and decrease the π-π interaction between ssDNA and SWCNTs, resulting in a certain amount of ssDNA dissociating from the complexes. The separated SWCNTs were selectively assembled on the self-assembled monolayer (SAM) modified Au electrode. Then the SWCNTs onto the SAM-modified Au electrode substantially restored heterogeneous electron transfer that was almost totally blocked by the SAM. The assembled SWCNTs could generate a considerably sensitive and specific tactic for signal transduction, which was related to the concentration of the arsenite. Through detecting the currents mediated by SWCNTs, a linear response to concentration of arsenite ranging from 0.5 to 10ppb and a detection limit of 0.5ppb was readily achieved with desirable specificity and sensitivity. Such a SWCNTs-based biosensor creates a simple, sensitive, nonradioactive route for detection of arsenite. In addition, this demonstration provides a new approach to fabrication of stable biosensors with favorable electrochemical properties believed to be appealing to electroanalytical applications. Copyright © 2015 Elsevier B.V. All rights reserved.
Single-Molecule Detection in Micron-Sized Capillaries
NASA Astrophysics Data System (ADS)
Ball, David A.; Shen, Guoqing; Davis, Lloyd M.
2004-11-01
The detection of individual molecules in solution by laser-induced fluorescence is becoming an increasingly important tool for biophysics research and biotechnology applications. In a typical single-molecule detection (SMD) experiment, diffusion is the dominant mode of transport of fluorophores through the focused laser beam. In order to more rapidly process a large number of slowly diffusing bio-molecules for applications in pharmaceutical drug discovery, a flow can be introduced within a capillary. If the flow speed is sufficient, bio-molecules will be carried through the probe volume significantly faster than by diffusion alone. Here we discuss SMD near the tip of, and in, such micron-sized capillaries, with a high numerical-aperture microscope objective used for confocal-epi-illumination along the axis of the capillary. Problems such as molecular adsorption to the glass are also addressed.
Microfluidic platform for multiplexed detection in single cells and methods thereof
DOE Office of Scientific and Technical Information (OSTI.GOV)
Wu, Meiye; Singh, Anup K.
The present invention relates to a microfluidic device and platform configured to conduct multiplexed analysis within the device. In particular, the device allows multiple targets to be detected on a single-cell level. Also provided are methods of performing multiplexed analyses to detect one or more target nucleic acids, proteins, and post-translational modifications.
Nanopore arrays in a silicon membrane for parallel single-molecule detection: DNA translocation
NASA Astrophysics Data System (ADS)
Zhang, Miao; Schmidt, Torsten; Jemt, Anders; Sahlén, Pelin; Sychugov, Ilya; Lundeberg, Joakim; Linnros, Jan
2015-08-01
Optical nanopore sensing offers great potential in single-molecule detection, genotyping, or DNA sequencing for high-throughput applications. However, one of the bottle-necks for fluorophore-based biomolecule sensing is the lack of an optically optimized membrane with a large array of nanopores, which has large pore-to-pore distance, small variation in pore size and low background photoluminescence (PL). Here, we demonstrate parallel detection of single-fluorophore-labeled DNA strands (450 bps) translocating through an array of silicon nanopores that fulfills the above-mentioned requirements for optical sensing. The nanopore array was fabricated using electron beam lithography and anisotropic etching followed by electrochemical etching resulting in pore diameters down to ∼7 nm. The DNA translocation measurements were performed in a conventional wide-field microscope tailored for effective background PL control. The individual nanopore diameter was found to have a substantial effect on the translocation velocity, where smaller openings slow the translocation enough for the event to be clearly detectable in the fluorescence. Our results demonstrate that a uniform silicon nanopore array combined with wide-field optical detection is a promising alternative with which to realize massively-parallel single-molecule detection.
Ma, Tracey; Byrne, Patrick A; Haya, Maryam; Elzohairy, Yoassry
2015-12-01
In 1998, Ontario implemented a remedial program called "Back On Track" (BOT) for individuals convicted of alcohol-impaired driving. Drivers convicted before October 2000 were exposed to a single-component program ("Edu BOT"); those convicted after participated in a multi-component program ("Full BOT"). We evaluated the impact of BOT, and the preceding 90-day roadside licence suspension, on drinking and driving recidivism, an outcome yet to be examined, using population-wide driver records. A Chi Square Test was used to compare the three-year cumulative incidence of recidivism between three historically-defined cohorts: No BOT, Edu BOT, and Full BOT. Stratified analyses by completion status and by age were also conducted. Analyses of the roadside suspension were conducted using an interrupted time series approach based on segmented Poisson/negative binomial regression. The roadside suspension was associated with a 65.2% reduction in drinking driving recidivism. In combination with indefinite suspensions for non-completion, the BOT program was also associated with a 21% decrease in drinking and driving recidivism in the three years following a CCC driving prohibition, from 8.5% to 6.7%. This reduction cannot be explained by pre-existing trends in recidivism. Conversion of the BOT program from the single-component version to the multi-component program further reduced the three-year cumulative incidence of recidivism to 5.5% (a total reduction of 35% from pre-BOT). Results provide strong converging evidence that remedial alcohol education/treatment programs in combination with other sanctions can produce substantial increases in road safety. Crown Copyright © 2015. Published by Elsevier Ltd. All rights reserved.
A rapid single-tube protocol for HAV detection by nested real-time PCR.
Hu, Yuan; Arsov, Ivica
2014-09-01
Infections by food-borne viruses such as hepatitis A virus (HAV) and norovirus are significant public health concerns worldwide. Since food-borne viruses are rarely confirmed through direct isolation from contaminated samples, highly sensitive molecular techniques remain the methods of choice for the detection of viral genetic material. Our group has previously developed a specific nested real-time PCR (NRT-PCR) assay for HAV detection that improved overall sensitivity. Furthermore in this study, we have developed a single-tube NRT-PCR approach for HAV detection in food samples that reduces the likelihood of cross contamination between tubes during sample manipulation. HAV RNA was isolated from HAV-spiked food samples and HAV-infected cell cultures. All reactions following HAV RNA isolation, including conventional reverse transcriptase PCR, nested-PCR, and RT-PCR were performed in a single tube. Our results demonstrated that all the samples tested positive by RT-PCR and nested-PCR were also positive by a single-tube NRT-PCR. The detection limits observed for HAV-infected cell cultures and HAV-spiked green onions were 0.1 and 1 PFU, respectively. This novel method retained the specificity and robustness of the original NRT-PCR method, while greatly reducing sample manipulation, turnaround time, and the risk of carry-over contamination. Single-tube NRT-PCR thus represents a promising new tool that can potentially facilitate the detection of HAV in foods thereby improving food safety and public health.
Efficient single photon detection by quantum dot resonant tunneling diodes.
Blakesley, J C; See, P; Shields, A J; Kardynał, B E; Atkinson, P; Farrer, I; Ritchie, D A
2005-02-18
We demonstrate that the resonant tunnel current through a double-barrier structure is sensitive to the capture of single photoexcited holes by an adjacent layer of quantum dots. This phenomenon could allow the detection of single photons with low dark count rates and high quantum efficiencies. The magnitude of the sensing current may be controlled via the thickness of the tunnel barriers. Larger currents give improved signal to noise and allow sub-mus photon time resolution.
Single-Electron Detection and Spectroscopy via Relativistic Cyclotron Radiation.
Asner, D M; Bradley, R F; de Viveiros, L; Doe, P J; Fernandes, J L; Fertl, M; Finn, E C; Formaggio, J A; Furse, D; Jones, A M; Kofron, J N; LaRoque, B H; Leber, M; McBride, E L; Miller, M L; Mohanmurthy, P; Monreal, B; Oblath, N S; Robertson, R G H; Rosenberg, L J; Rybka, G; Rysewyk, D; Sternberg, M G; Tedeschi, J R; Thümmler, T; VanDevender, B A; Woods, N L
2015-04-24
It has been understood since 1897 that accelerating charges must emit electromagnetic radiation. Although first derived in 1904, cyclotron radiation from a single electron orbiting in a magnetic field has never been observed directly. We demonstrate single-electron detection in a novel radio-frequency spectrometer. The relativistic shift in the cyclotron frequency permits a precise electron energy measurement. Precise beta electron spectroscopy from gaseous radiation sources is a key technique in modern efforts to measure the neutrino mass via the tritium decay end point, and this work demonstrates a fundamentally new approach to precision beta spectroscopy for future neutrino mass experiments.
DNAzyme based gap-LCR detection of single-nucleotide polymorphism.
Zhou, Li; Du, Feng; Zhao, Yongyun; Yameen, Afshan; Chen, Haodong; Tang, Zhuo
2013-07-15
Fast and accurate detection of single-nucleotide polymorphism (SNP) is thought more and more important for understanding of human physiology and elucidating the molecular based diseases. A great deal of effort has been devoted to developing accurate, rapid, and cost-effective technologies for SNP analysis. However most of those methods developed to date incorporate complicated probe labeling and depend on advanced equipment. The DNAzyme based Gap-LCR detection method averts any chemical modification on probes and circumvents those problems by incorporating a short functional DNA sequence into one of LCR primers. Two kinds of exonuclease are utilized in our strategy to digest all the unreacted probes and release the DNAzymes embedded in the LCR product. The DNAzyme applied in our method is a versatile tool to report the result of SNP detection in colorimetric or fluorometric ways for different detection purposes. Copyright © 2013 Elsevier B.V. All rights reserved.
Detecting earthquakes over a seismic network using single-station similarity measures
NASA Astrophysics Data System (ADS)
Bergen, Karianne J.; Beroza, Gregory C.
2018-06-01
New blind waveform-similarity-based detection methods, such as Fingerprint and Similarity Thresholding (FAST), have shown promise for detecting weak signals in long-duration, continuous waveform data. While blind detectors are capable of identifying similar or repeating waveforms without templates, they can also be susceptible to false detections due to local correlated noise. In this work, we present a set of three new methods that allow us to extend single-station similarity-based detection over a seismic network; event-pair extraction, pairwise pseudo-association, and event resolution complete a post-processing pipeline that combines single-station similarity measures (e.g. FAST sparse similarity matrix) from each station in a network into a list of candidate events. The core technique, pairwise pseudo-association, leverages the pairwise structure of event detections in its network detection model, which allows it to identify events observed at multiple stations in the network without modeling the expected moveout. Though our approach is general, we apply it to extend FAST over a sparse seismic network. We demonstrate that our network-based extension of FAST is both sensitive and maintains a low false detection rate. As a test case, we apply our approach to 2 weeks of continuous waveform data from five stations during the foreshock sequence prior to the 2014 Mw 8.2 Iquique earthquake. Our method identifies nearly five times as many events as the local seismicity catalogue (including 95 per cent of the catalogue events), and less than 1 per cent of these candidate events are false detections.
Detecting Earthquakes over a Seismic Network using Single-Station Similarity Measures
NASA Astrophysics Data System (ADS)
Bergen, Karianne J.; Beroza, Gregory C.
2018-03-01
New blind waveform-similarity-based detection methods, such as Fingerprint and Similarity Thresholding (FAST), have shown promise for detecting weak signals in long-duration, continuous waveform data. While blind detectors are capable of identifying similar or repeating waveforms without templates, they can also be susceptible to false detections due to local correlated noise. In this work, we present a set of three new methods that allow us to extend single-station similarity-based detection over a seismic network; event-pair extraction, pairwise pseudo-association, and event resolution complete a post-processing pipeline that combines single-station similarity measures (e.g. FAST sparse similarity matrix) from each station in a network into a list of candidate events. The core technique, pairwise pseudo-association, leverages the pairwise structure of event detections in its network detection model, which allows it to identify events observed at multiple stations in the network without modeling the expected move-out. Though our approach is general, we apply it to extend FAST over a sparse seismic network. We demonstrate that our network-based extension of FAST is both sensitive and maintains a low false detection rate. As a test case, we apply our approach to two weeks of continuous waveform data from five stations during the foreshock sequence prior to the 2014 Mw 8.2 Iquique earthquake. Our method identifies nearly five times as many events as the local seismicity catalog (including 95% of the catalog events), and less than 1% of these candidate events are false detections.
Single Cell Detection with Driven Magnetic Beads
NASA Astrophysics Data System (ADS)
McNaughton, B. H.; Agayan, R. R.; Stoica, V. A.; Clarke, R.; Kopelman, R.
Shifts in the nonlinear rotational frequency of magnetic beads (microspheres) offer a new and dynamic approach for the detection of single cells. We present the first demonstration of this capability by measuring the changes in the nonlinear rotational frequency of magnetic beads driven by an external magnetic field. The presence of an Escherichia coli bacterium on the surface of a 2.0 μm magnetic bead affects the drag of the system, thus changing the nonlinear rotation rate. Measurement of this rotational frequency is straight-forward utilizing standard microscopy techniques.
Two particle tracking and detection in a single Gaussian beam optical trap.
Praveen, P; Yogesha; Iyengar, Shruthi S; Bhattacharya, Sarbari; Ananthamurthy, Sharath
2016-01-20
We have studied in detail the situation wherein two microbeads are trapped axially in a single-beam Gaussian intensity profile optical trap. We find that the corner frequency extracted from a power spectral density analysis of intensity fluctuations recorded on a quadrant photodetector (QPD) is dependent on the detection scheme. Using forward- and backscattering detection schemes with single and two laser wavelengths along with computer simulations, we conclude that fluctuations detected in backscattering bear true position information of the bead encountered first in the beam propagation direction. Forward scattering, on the other hand, carries position information of both beads with substantial contribution from the bead encountered first along the beam propagation direction. Mie scattering analysis further reveals that the interference term from the scattering of the two beads contributes significantly to the signal, precluding the ability to resolve the positions of the individual beads in forward scattering. In QPD-based detection schemes, detection through backscattering, thereby, is imperative to track the true displacements of axially trapped microbeads for possible studies on light-mediated interbead interactions.
Single-molecule two-colour coincidence detection to probe biomolecular associations.
Orte, Angel; Clarke, Richard; Klenerman, David
2010-08-01
Two-colour coincidence detection (TCCD) is a form of single-molecule fluorescence developed to sensitively detect and characterize associated biomolecules without any separation, in solution, on the cell membrane and in live cells. In the present short review, we first explain the principles of the method and then describe the application of TCCD to a range of biomedical problems and how this method may be developed further in the future to try to monitor biological processes in live cells.
Detection of gas molecules on single Mn adatom adsorbed graphyne: a DFT-D study
NASA Astrophysics Data System (ADS)
Lu, Zhansheng; Lv, Peng; Ma, Dongwei; Yang, Xinwei; Li, Shuo; Yang, Zongxian
2018-02-01
As one of the prominent applications in intelligent systems, gas sensing technology has attracted great interest in both industry and academia. In the current study, the pristine graphyne (GY) without and with a single Mn atom is investigated to detect the gas molecules (CO, CH4, CO2, NH3, NO and O2). The pristine GY is promising to detect O2 molecules because of its chemical adsorption on GY with large electron transfer. The great stability of the Mn/GY is found, and the Mn atom prefers to anchor at the alkyne ring as a single atom. Upon single Mn atom anchoring, the sensitivity and selectivity of GY based gas sensors is significantly improved for various molecules, except CH4. The recovery time of the Mn/GY after detecting the gas molecules may help to appraise the detection efficiency for the Mn/GY. The current study will help to understand the mechanism of detecting the gas molecules, and extend the potentially fascinating applications of GY-based materials.
Single Molecule Electrochemical Detection in Aqueous Solutions and Ionic Liquids.
Byers, Joshua C; Paulose Nadappuram, Binoy; Perry, David; McKelvey, Kim; Colburn, Alex W; Unwin, Patrick R
2015-10-20
Single molecule electrochemical detection (SMED) is an extremely challenging aspect of electroanalytical chemistry, requiring unconventional electrochemical cells and measurements. Here, SMED is reported using a "quad-probe" (four-channel probe) pipet cell, fabricated by depositing carbon pyrolytically into two diagonally opposite barrels of a laser-pulled quartz quadruple-barreled pipet and filling the open channels with electrolyte solution, and quasi-reference counter electrodes. A meniscus forms at the end of the probe covering the two working electrodes and is brought into contact with a substrate working electrode surface. In this way, a nanogap cell is produced whereby the two carbon electrodes in the pipet can be used to promote redox cycling of an individual molecule with the substrate. Anticorrelated currents generated at the substrate and tip electrodes, at particular distances (typically tens of nanometers), are consistent with the detection of single molecules. The low background noise realized in this droplet format opens up new opportunities in single molecule electrochemistry, including the use of ionic liquids, as well as aqueous solution, and the quantitative assessment and analysis of factors influencing redox cycling currents, due to a precisely known gap size.
Single- and multi-frequency detection of surface displacements via scanning probe microscopy.
Romanyuk, Konstantin; Luchkin, Sergey Yu; Ivanov, Maxim; Kalinin, Arseny; Kholkin, Andrei L
2015-02-01
Piezoresponse force microscopy (PFM) provides a novel opportunity to detect picometer-level displacements induced by an electric field applied through a conducting tip of an atomic force microscope (AFM). Recently, it was discovered that superb vertical sensitivity provided by PFM is high enough to monitor electric-field-induced ionic displacements in solids, the technique being referred to as electrochemical strain microscopy (ESM). ESM has been implemented only in multi-frequency detection modes such as dual AC resonance tracking (DART) and band excitation, where the response is recorded within a finite frequency range, typically around the first contact resonance. In this paper, we analyze and compare signal-to-noise ratios of the conventional single-frequency method with multi-frequency regimes of measuring surface displacements. Single-frequency detection ESM is demonstrated using a commercial AFM.
Nonlinear ultrasonic fatigue crack detection using a single piezoelectric transducer
NASA Astrophysics Data System (ADS)
An, Yun-Kyu; Lee, Dong Jun
2016-04-01
This paper proposes a new nonlinear ultrasonic technique for fatigue crack detection using a single piezoelectric transducer (PZT). The proposed technique identifies a fatigue crack using linear (α) and nonlinear (β) parameters obtained from only a single PZT mounted on a target structure. Based on the different physical characteristics of α and β, a fatigue crack-induced feature is able to be effectively isolated from the inherent nonlinearity of a target structure and data acquisition system. The proposed technique requires much simpler test setup and less processing costs than the existing nonlinear ultrasonic techniques, but fast and powerful. To validate the proposed technique, a real fatigue crack is created in an aluminum plate, and then false positive and negative tests are carried out under varying temperature conditions. The experimental results reveal that the fatigue crack is successfully detected, and no positive false alarm is indicated.
Ultrasensitive Detection of Ricin Toxin in Multiple Sample Matrixes Using Single-Domain Antibodies.
Gaylord, Shonda T; Dinh, Trinh L; Goldman, Ellen R; Anderson, George P; Ngan, Kevin C; Walt, David R
2015-07-07
Ricin is an extremely potent ribosomal inactivating protein listed as a Category B select agent. Although ricin intoxication is not transmittable from person to person, even a single ricin molecule can lead to cell necrosis because it inactivates 1500 ribosomes/min. Since there is currently no vaccine or therapeutic treatment for ricin intoxication, ultrasensitive analytical assays capable of detecting ricin in a variety of matrixes are urgently needed to limit exposure to individuals as well as communities. In this paper, we present the development and application of a single-molecule array (Simoa) for the detection of ricin toxin in human urine and serum. Single-domain antibodies (sdAbs), among the smallest engineered binding fragments, were chemically coupled to the surface of paramagnetic beads for the sensitive detection of ricin toxin. The Simoa was able to detect ricin at levels of 10 fg/mL, 100 fg/mL, and 1 pg/mL in buffer, urine and serum, respectively, in a fraction of the assay time need using immuno-polymerase chain reaction (IPCR). Using a fully automated state-of-the-art platform, the Simoa HD-1 analyzer, the assay time was reduced to 64 min.
NASA Astrophysics Data System (ADS)
Takakura, T.; Yanagi, I.; Goto, Y.; Ishige, Y.; Kohara, Y.
2016-03-01
We developed a resistive-pulse sensor with a solid-state pore and measured the latex agglutination of submicron particles induced by antigen-antibody interaction for single-molecule detection of proteins. We fabricated the pore based on numerical simulation to clearly distinguish between monomer and dimer latex particles. By measuring single dimers agglutinated in the single-molecule regime, we detected single human alpha-fetoprotein molecules. Adjusting the initial particle concentration improves the limit of detection (LOD) to 95 fmol/l. We established a theoretical model of the LOD by combining the reaction kinetics and the counting statistics to explain the effect of initial particle concentration on the LOD. The theoretical model shows how to improve the LOD quantitatively. The single-molecule detection studied here indicates the feasibility of implementing a highly sensitive immunoassay by a simple measurement method using resistive-pulse sensing.
Biomolecule detection based on Si single-electron transistors for practical use
NASA Astrophysics Data System (ADS)
Nakajima, Anri; Kudo, Takashi; Furuse, Sadaharu
2013-07-01
Experimental and theoretical analyses demonstrated that ultra-sensitive biomolecule detection can be achieved using a Si single-electron transistor (SET). A multi-island channel structure was used to enable room-temperature operation. Coulomb oscillation increases transconductance without increasing channel width, which increases detection sensitivity to a charged target. A biotin-modified SET biosensor was used to detect streptavidin at a dilute concentration. In addition, an antibody-functionalized SET biosensor was used for immunodetection of prostate-specific antigen, demonstrating its suitability for practical use. The feasibility of ultra-sensitive detection of biomolecules for practical use by using a SET biosensor was clearly proven through this systematic study.
Single molecule fluorescence burst detection of DNA fragments separated by capillary electrophoresis
DOE Office of Scientific and Technical Information (OSTI.GOV)
Haab, B.B.; Mathies, R.A.
A method has been developed for detecting DNA separated by capillary gel electrophoresis (CGE) using single molecule photon burst counting. A confocal fluorescence microscope was used to observe the fluorescence bursts from single molecules of DNA multiply labeled with the thiazole orange derivative TO6 as they passed through the nearly 2-{mu}m diameter focused laser beam. Amplified photo-electron pulses from the photomultiplier are grouped into bins of 360-450 {mu}s in duration, and the resulting histogram is stored in a computer for analysis. Solutions of M13 DNA were first flowed through the capillary at various concentrations, and the resulting data were usedmore » to optimize the parameters for digital filtering using a low-pass Fourier filter, selecting a discriminator level for peak detection, and applying a peak-calling algorithm. The optimized single molecule counting method was then applied to an electrophoretic separation of M13 DNA and to a separation of pBR 322 DNA from pRL 277 DNA. Clusters of discreet fluorescence bursts were observed at the expected appearance time of each DNA band. The auto-correlation function of these data indicated transit times that were consistent with the observed electrophoretic velocity. These separations were easily detected when only 50-100 molecules of DNA per band traveled through the detection region. This new detection technology should lead to the routine analysis of DNA in capillary columns with an on-column sensitivity of nearly 100 DNA molecules/band or better. 45 refs., 10 figs.« less
Song, Yunke; Zhang, Yi; Wang, Tza-Huei
2013-04-08
Gene point mutations present important biomarkers for genetic diseases. However, existing point mutation detection methods suffer from low sensitivity, specificity, and a tedious assay processes. In this report, an assay technology is proposed which combines the outstanding specificity of gap ligase chain reaction (Gap-LCR), the high sensitivity of single-molecule coincidence detection, and the superior optical properties of quantum dots (QDs) for multiplexed detection of point mutations in genomic DNA. Mutant-specific ligation products are generated by Gap-LCR and subsequently captured by QDs to form DNA-QD nanocomplexes that are detected by single-molecule spectroscopy (SMS) through multi-color fluorescence burst coincidence analysis, allowing for multiplexed mutation detection in a separation-free format. The proposed assay is capable of detecting zeptomoles of KRAS codon 12 mutation variants with near 100% specificity. Its high sensitivity allows direct detection of KRAS mutation in crude genomic DNA without PCR pre-amplification. Copyright © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Nano-Electromechanical Systems: Displacement Detection and the Mechanical Single Electron Shuttle
NASA Astrophysics Data System (ADS)
Blick, R. H.; Beil, F. W.; Höhberger, E.; Erbe, A.; Weiss, C.
For an introduction to nano-electromechanical systems we present measurements on nanomechanical resonators operating in the radio frequency range. We discuss in detail two different schemes of displacement detection for mechanical resonators, namely conventional reflection measurements of a probing signal and direct detection by capacitive coupling via a gate electrode. For capacitive detection we employ an on-chip preamplifier, which enables direct measurements of the resonator's disp lacement. We observe that the mechanical quality factor of the resonator depends on the detection technique applied, which is verified in model calculations and report on the detection of sub-harmonics. In the second part we extend our investigations to include transport of single electrons through an electron island on the tip of a nanomachined mechanical pendulum. The pendulum is operated by applying a modulating electromagnetic field in the range of 1 - 200 MHz, leading to mechanical oscillations between two laterally integrated source and drain contacts. Forming tunneling barriers the metallic tip shuttles single electrons from source to drain. The resulting tunneling current shows distinct features corresponding to the discrete mechanical eigenfrequencies of the pendulum. We report on measurements covering the temperature range from 300 K down to 4.2 K. The transport properties of the device are compared in detail to model calculations based on a Master-equation approach.
Zheng, Yuanhui; Soeriyadi, Alexander H.; Rosa, Lorenzo; Ng, Soon Hock; Bach, Udo; Justin Gooding, J.
2015-01-01
Single-molecule surface-enhanced Raman spectroscopy (SERS) has attracted increasing interest for chemical and biochemical sensing. Many conventional substrates have a broad distribution of SERS enhancements, which compromise reproducibility and result in slow response times for single-molecule detection. Here we report a smart plasmonic sensor that can reversibly trap a single molecule at hotspots for rapid single-molecule detection. The sensor was fabricated through electrostatic self-assembly of gold nanoparticles onto a gold/silica-coated silicon substrate, producing a high yield of uniformly distributed hotspots on the surface. The hotspots were isolated with a monolayer of a thermoresponsive polymer (poly(N-isopropylacrylamide)), which act as gates for molecular trapping at the hotspots. The sensor shows not only a good SERS reproducibility but also a capability to repetitively trap and release molecules for single-molecular sensing. The single-molecule sensitivity is experimentally verified using SERS spectral blinking and bianalyte methods. PMID:26549539
Sansinena, Jose-Maria [Los Alamos, NM; Redondo, Antonio [Los Alamos, NM; Olazabal, Virginia [Los Alamos, NM; Hoffbauer, Mark A [Los Alamos, NM; Akhadov, Elshan A [Los Alamos, NM
2009-12-29
A barrier structure for use in an electrochemical stochastic membrane sensor for single molecule detection. The sensor is based upon inorganic nanopores having electrically tunable dimensions. The inorganic nanopores are formed from inorganic materials and an electrically conductive polymer. Methods of making the barrier structure and sensing single molecules using the barrier structure are also described.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Sansinena, Jose-Maria; Redondo, Antonio; Olazabal, Virginia
2017-09-12
A barrier structure for use in an electrochemical stochastic membrane sensor for single molecule detection. The sensor is based upon inorganic nanopores having electrically tunable dimensions. The inorganic nanopores are formed from inorganic materials and an electrically conductive polymer. Methods of making the barrier structure and sensing single molecules using the barrier structure are also described.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Sansinena, Jose-Maria; Redondo, Antonio; Olazabal, Virginia
2017-07-18
A barrier structure for use in an electrochemical stochastic membrane sensor for single molecule detection. The sensor is based upon inorganic nanopores having electrically tunable dimensions. The inorganic nanopores are formed from inorganic materials and an electrically conductive polymer. Methods of making the barrier structure and sensing single molecules using the barrier structure are also described.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Sansinena, Jose-Maria; Redondo, Antonio; Olazabal, Virginia
A barrier structure for use in an electrochemical stochastic membrane sensor for single molecule detection. The sensor is based upon inorganic nanopores having electrically tunable dimensions. The inorganic nanopores are formed from inorganic materials and an electrically conductive polymer. Methods of making the barrier structure and sensing single molecules using the barrier structure are also described.
Combining nanofluidics and plasmonics for single molecule detection
NASA Astrophysics Data System (ADS)
West, Melanie M.
Single molecule detection is limited by the small scattering cross-section of molecules which leads to weak optical signals that can be obscured by background noise. The combination of plasmonics and nanofluidics in an integrated nano-device has the potential to provide the signal enhancement necessary for the detection of single molecules. The purpose of this investigation was to optimize the fabrication of an optofluidic device that integrates a nanochannel with a plasmonic bowtie antenna. The fluidic structure of the device was fabricated using UV-nanoimprint lithography, and the gold plasmonic antennas were fabricated using a shadow evaporation and lift-off process. The effect of electron beam lithography doses on the resolution of antenna-nanochannel configurations was studied to minimize antenna gap size while maintaining the integrity of the imprinted features. The smallest antenna gap size that was achieved was 46 nm. The antennas were characterized using dark field spectroscopy to find the resonance shift, which indicated the appropriate range for optical signal enhancement. The dark field scattering results showed antennas with a broad and well-defined resonance shift that ranged from 650--800 nm. The Raman scattering results showed the highest enhancement factor (EF = 2) for antennas with an "inverted configuration," which involved having the triangles of the antenna facing back-to-back rather than the more conventional tip-to-tip bowtie arrangement.
Single-trial lie detection using a combined fNIRS-polygraph system
Bhutta, M. Raheel; Hong, Melissa J.; Kim, Yun-Hee; Hong, Keum-Shik
2015-01-01
Deception is a human behavior that many people experience in daily life. It involves complex neuronal activities in addition to several physiological changes in the body. A polygraph, which can measure some of the physiological responses from the body, has been widely employed in lie-detection. Many researchers, however, believe that lie detection can become more precise if the neuronal changes that occur in the process of deception can be isolated and measured. In this study, we combine both measures (i.e., physiological and neuronal changes) for enhanced lie-detection. Specifically, to investigate the deception-related hemodynamic response, functional near-infrared spectroscopy (fNIRS) is applied at the prefrontal cortex besides a commercially available polygraph system. A mock crime scenario with a single-trial stimulus is set up as a deception protocol. The acquired data are classified into “true” and “lie” classes based on the fNIRS-based hemoglobin-concentration changes and polygraph-based physiological signal changes. Linear discriminant analysis is utilized as a classifier. The results indicate that the combined fNIRS-polygraph system delivers much higher classification accuracy than that of a singular system. This study demonstrates a plausible solution toward single-trial lie-detection by combining fNIRS and the polygraph. PMID:26082733
On-chip detection of non-classical light by scalable integration of single-photon detectors
Najafi, Faraz; Mower, Jacob; Harris, Nicholas C.; Bellei, Francesco; Dane, Andrew; Lee, Catherine; Hu, Xiaolong; Kharel, Prashanta; Marsili, Francesco; Assefa, Solomon; Berggren, Karl K.; Englund, Dirk
2015-01-01
Photonic-integrated circuits have emerged as a scalable platform for complex quantum systems. A central goal is to integrate single-photon detectors to reduce optical losses, latency and wiring complexity associated with off-chip detectors. Superconducting nanowire single-photon detectors (SNSPDs) are particularly attractive because of high detection efficiency, sub-50-ps jitter and nanosecond-scale reset time. However, while single detectors have been incorporated into individual waveguides, the system detection efficiency of multiple SNSPDs in one photonic circuit—required for scalable quantum photonic circuits—has been limited to <0.2%. Here we introduce a micrometer-scale flip-chip process that enables scalable integration of SNSPDs on a range of photonic circuits. Ten low-jitter detectors are integrated on one circuit with 100% device yield. With an average system detection efficiency beyond 10%, and estimated on-chip detection efficiency of 14–52% for four detectors operated simultaneously, we demonstrate, to the best of our knowledge, the first on-chip photon correlation measurements of non-classical light. PMID:25575346
Liu, Meiying; Yuan, Min; Lou, Xinhui; Mao, Hongju; Zheng, Dongmei; Zou, Ruxing; Zou, Nengli; Tang, Xiangrong; Zhao, Jianlong
2011-07-15
We report here an optical approach that enables highly selective and colorimetric single-base mismatch detection without the need of target modification, precise temperature control or stringent washes. The method is based on the finding that nucleoside monophosphates (dNMPs), which are digested elements of DNA, can better stabilize unmodified gold nanoparticles (AuNPs) than single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA) with the same base-composition and concentration. The method combines the exceptional mismatch discrimination capability of the structure-selective nucleases with the attractive optical property of AuNPs. Taking S1 nuclease as one example, the perfectly matched 16-base synthetic DNA target was distinctively differentiated from those with single-base mutation located at any position of the 16-base synthetic target. Single-base mutations present in targets with varied length up to 80-base, located either in the middle or near to the end of the targets, were all effectively detected. In order to prove that the method can be potentially used for real clinic samples, the single-base mismatch detections with two HBV genomic DNA samples were conducted. To further prove the generality of this method and potentially overcome the limitation on the detectable lengths of the targets of the S1 nuclease-based method, we also demonstrated the use of a duplex-specific nuclease (DSN) for color reversed single-base mismatch detection. The main limitation of the demonstrated methods is that it is limited to detect mutations in purified ssDNA targets. However, the method coupled with various convenient ssDNA generation and purification techniques, has the potential to be used for the future development of detector-free testing kits in single nucleotide polymorphism screenings for disease diagnostics and treatments. Copyright © 2011 Elsevier B.V. All rights reserved.
A theoretical investigation of single-molecule fluorescence detection on thin metallic layers.
Enderlein, J
2000-04-01
In the present paper, the excitation and detection of single-molecule fluorescence over thin metallic films is studied theoretically within the framework of classical electrodynamics. The model takes into account the specific conditions of surface plasmon-assisted optical excitation, fluorescence quenching by the metal film, and detection geometry. Extensive numerical results are presented for gold, silver, and aluminum films, showing the detectable fluorescence intensities and their dependence on film thickness and the fluorescent molecule's position under optimal excitation conditions.
[Single-molecule detection and characterization of DNA replication based on DNA origami].
Wang, Qi; Fan, Youjie; Li, Bin
2014-08-01
To investigate single-molecule detection and characterization of DNA replication. Single-stranded DNA (ssDNA) as the template of DNA replication was attached to DNA origami by a hybridization reaction based on the complementary base-pairing principle. DNA replication catalyzed by E.coli DNA polymerase I Klenow Fragment (KF) was detected using atomic force microscopy (AFM). The height variations between the ssDNA and the double-stranded DNA (dsDNA), the distribution of KF during DNA replication and biotin-streptavidin (BA) complexes on the DNA strand after replication were detected. Agarose gel electrophoresis was employed to analyze the changes in the DNA after replication. The designed ssDNA could be anchored on the target positions of over 50% of the DNA origami. The KF was capable of binding to the ssDNA fixed on DNA origami and performing its catalytic activities, and was finally dissociated from the DNA after replication. The height of DNA strand increased by about 0.7 nm after replication. The addition of streptavidin also resulted in an DNA height increase to about 4.9 nm due to the formation of BA complexes on the biotinylated dsDNA. The resulting dsDNA and BA complex were subsequently confirmed by agarose gel electrophoresis. The combination of AFM and DNA origami allows detection and characterization of DNA replication at the single molecule level, and this approach provides better insights into the mechanism of DNA polymerase and the factors affecting DNA replication.
Shah, Sheel; Lubeck, Eric; Schwarzkopf, Maayan; He, Ting-Fang; Greenbaum, Alon; Sohn, Chang Ho; Lignell, Antti; Choi, Harry M T; Gradinaru, Viviana; Pierce, Niles A; Cai, Long
2016-08-01
Accurate and robust detection of mRNA molecules in thick tissue samples can reveal gene expression patterns in single cells within their native environment. Preserving spatial relationships while accessing the transcriptome of selected cells is a crucial feature for advancing many biological areas - from developmental biology to neuroscience. However, because of the high autofluorescence background of many tissue samples, it is difficult to detect single-molecule fluorescence in situ hybridization (smFISH) signals robustly in opaque thick samples. Here, we draw on principles from the emerging discipline of dynamic nucleic acid nanotechnology to develop a robust method for multi-color, multi-RNA imaging in deep tissues using single-molecule hybridization chain reaction (smHCR). Using this approach, single transcripts can be imaged using epifluorescence, confocal or selective plane illumination microscopy (SPIM) depending on the imaging depth required. We show that smHCR has high sensitivity in detecting mRNAs in cell culture and whole-mount zebrafish embryos, and that combined with SPIM and PACT (passive CLARITY technique) tissue hydrogel embedding and clearing, smHCR can detect single mRNAs deep within thick (0.5 mm) brain slices. By simultaneously achieving ∼20-fold signal amplification and diffraction-limited spatial resolution, smHCR offers a robust and versatile approach for detecting single mRNAs in situ, including in thick tissues where high background undermines the performance of unamplified smFISH. © 2016. Published by The Company of Biologists Ltd.
Detection of confinement and jumps in single-molecule membrane trajectories
NASA Astrophysics Data System (ADS)
Meilhac, N.; Le Guyader, L.; Salomé, L.; Destainville, N.
2006-01-01
We propose a variant of the algorithm by [R. Simson, E. D. Sheets, and K. Jacobson, Biophys. 69, 989 (1995)]. Their algorithm was developed to detect transient confinement zones in experimental single-particle tracking trajectories of diffusing membrane proteins or lipids. We show that our algorithm is able to detect confinement in a wider class of confining potential shapes than that of Simson Furthermore, it enables to detect not only temporary confinement but also jumps between confinement zones. Jumps are predicted by membrane skeleton fence and picket models. In the case of experimental trajectories of μ -opioid receptors, which belong to the family of G-protein-coupled receptors involved in a signal transduction pathway, this algorithm confirms that confinement cannot be explained solely by rigid fences.
A Lateral Flow Biosensor for the Detection of Single Nucleotide Polymorphisms.
Zeng, Lingwen; Xiao, Zhuo
2017-01-01
A lateral flow biosensor (LFB) is introduced for the detection of single nucleotide polymorphisms (SNPs). The assay is composed of two steps: circular strand displacement reaction and lateral flow biosensor detection. In step 1, the nucleotide at SNP site is recognized by T4 DNA ligase and the signal is amplified by strand displacement DNA polymerase, which can be accomplished at a constant temperature. In step 2, the reaction product of step 1 is detected by a lateral flow biosensor, which is a rapid and cost effective tool for nuclei acid detection. Comparing with conventional methods, it requires no complicated machines. It is suitable for the use of point of care diagnostics. Therefore, this simple, cost effective, robust, and promising LFB detection method of SNP has great potential for the detection of genetic diseases, personalized medicine, cancer related mutations, and drug-resistant mutations of infectious agents.
miRNA detection at single-cell resolution using microfluidic LNA flow-FISH
Wu, Meiye; Piccini, Matthew Ernest; Koh, Chung -Yan; ...
2014-08-20
Flow cytometry in combination with fluorescent in situ hybridization (flow-FISH) is a powerful technique that can be utilized to rapidly detect nucleic acids at single-cell resolution without the need for homogenization or nucleic acid extraction. Here, we describe a microfluidic-based method which enables the detection of microRNAs or miRNAs in single intact cells by flow-FISH using locked nucleic acid (LNA)-containing probes. Our method can be applied to all RNA species including mRNA and small noncoding RNA and is suitable for multiplexing with protein immunostaining in the same cell. For demonstration of our method, this chapter details the detection of miR155more » and CD69 protein in PMA and ionomycin-stimulated Jurkat cells. Here, we also include instructions on how to set up a microfluidic chip sample preparation station to prepare cells for imaging and analysis on a commercial flow cytometer or a custom-built micro-flow cytometer.« less
Iino, Ryota; Sakakihara, Shouichi; Matsumoto, Yoshimi; Nishino, Kunihiko
2016-01-01
A directly accessible femtoliter droplet array as a platform for single-cell detection and collection of persister bacteria is described. Device microfabrication, femtoliter droplet array formation and concomitant enclosure of single cells, long-term culture and observation of single cells in droplets, and collection of identified persisters from single droplets are described in detail.
Sevenler, Derin; Daaboul, George G; Ekiz Kanik, Fulya; Ünlü, Neşe Lortlar; Ünlü, M Selim
2018-05-21
DNA and protein microarrays are a high-throughput technology that allow the simultaneous quantification of tens of thousands of different biomolecular species. The mediocre sensitivity and limited dynamic range of traditional fluorescence microarrays compared to other detection techniques have been the technology's Achilles' heel and prevented their adoption for many biomedical and clinical diagnostic applications. Previous work to enhance the sensitivity of microarray readout to the single-molecule ("digital") regime have either required signal amplifying chemistry or sacrificed throughput, nixing the platform's primary advantages. Here, we report the development of a digital microarray which extends both the sensitivity and dynamic range of microarrays by about 3 orders of magnitude. This technique uses functionalized gold nanorods as single-molecule labels and an interferometric scanner which can rapidly enumerate individual nanorods by imaging them with a 10× objective lens. This approach does not require any chemical signal enhancement such as silver deposition and scans arrays with a throughput similar to commercial fluorescence scanners. By combining single-nanoparticle enumeration and ensemble measurements of spots when the particles are very dense, this system achieves a dynamic range of about 6 orders of magnitude directly from a single scan. As a proof-of-concept digital protein microarray assay, we demonstrated detection of hepatitis B virus surface antigen in buffer with a limit of detection of 3.2 pg/mL. More broadly, the technique's simplicity and high-throughput nature make digital microarrays a flexible platform technology with a wide range of potential applications in biomedical research and clinical diagnostics.
Detection of protease and protease activity using a single nanoscrescent SERS probe
Liu, Gang L.; Ellman, Jonathan A.; Lee, Luke P.; Chen, Fanqing Frank
2013-01-29
This invention pertains to the in vitro detection of proteases using a single peptide-conjugate nanocrescent surface enhanced Raman scattering (SERS) probes with at least nanomolar sensitivity. The probe enables detection of proteolytic activity in extremely small volume and at low concentration. In certain embodiments the probes comprise an indicator for the detection of an active protease, where the indicator comprises a nanocrescent attached to a peptide, where said peptide comprises a recognition site for the protease and a Raman tag attached to the peptide.
Detection of protease and protease activity using a single nanocrescent SERS probe
Liu, Gang L.; Ellman, Jonathan A.; Lee, Luke P.; Chen, Fanqing Frank
2015-09-29
This invention pertains to the in vitro detection of proteases using a single peptide-conjugate nanocrescent surface enhanced Raman scattering (SERS) probes with at least nanomolar sensitivity. The probe enables detection of proteolytic activity in extremely small volume and at low concentration. In certain embodiments the probes comprise an indicator for the detection of an active protease, where the indicator comprises a nanocrescent attached to a peptide, where said peptide comprises a recognition site for the protease and a Raman tag attached to the peptide.
Automatic detection and classification of artifacts in single-channel EEG.
Olund, Thomas; Duun-Henriksen, Jonas; Kjaer, Troels W; Sorensen, Helge B D
2014-01-01
Ambulatory EEG monitoring can provide medical doctors important diagnostic information, without hospitalizing the patient. These recordings are however more exposed to noise and artifacts compared to clinically recorded EEG. An automatic artifact detection and classification algorithm for single-channel EEG is proposed to help identifying these artifacts. Features are extracted from the EEG signal and wavelet subbands. Subsequently a selection algorithm is applied in order to identify the best discriminating features. A non-linear support vector machine is used to discriminate among different artifact classes using the selected features. Single-channel (Fp1-F7) EEG recordings are obtained from experiments with 12 healthy subjects performing artifact inducing movements. The dataset was used to construct and validate the model. Both subject-specific and generic implementation, are investigated. The detection algorithm yield an average sensitivity and specificity above 95% for both the subject-specific and generic models. The classification algorithm show a mean accuracy of 78 and 64% for the subject-specific and generic model, respectively. The classification model was additionally validated on a reference dataset with similar results.
Single molecule transistor based nanopore for the detection of nicotine
NASA Astrophysics Data System (ADS)
Ray, S. J.
2014-12-01
A nanopore based detection methodology was proposed and investigated for the detection of Nicotine. This technique uses a Single Molecular Transistor working as a nanopore operational in the Coulomb Blockade regime. When the Nicotine molecule is pulled through the nanopore area surrounded by the Source(S), Drain (D), and Gate electrodes, the charge stability diagram can detect the presence of the molecule and is unique for a specific molecular structure. Due to the weak coupling between the different electrodes which is set by the nanopore size, the molecular energy states stay almost unaffected by the electrostatic environment that can be realised from the charge stability diagram. Identification of different orientation and position of the Nicotine molecule within the nanopore area can be made from specific regions of overlap between different charge states on the stability diagram that could be used as an electronic fingerprint for detection. This method could be advantageous and useful to detect the presence of Nicotine in smoke which is usually performed using chemical chromatography techniques.
Single molecule transistor based nanopore for the detection of nicotine
DOE Office of Scientific and Technical Information (OSTI.GOV)
Ray, S. J., E-mail: ray.sjr@gmail.com
A nanopore based detection methodology was proposed and investigated for the detection of Nicotine. This technique uses a Single Molecular Transistor working as a nanopore operational in the Coulomb Blockade regime. When the Nicotine molecule is pulled through the nanopore area surrounded by the Source(S), Drain (D), and Gate electrodes, the charge stability diagram can detect the presence of the molecule and is unique for a specific molecular structure. Due to the weak coupling between the different electrodes which is set by the nanopore size, the molecular energy states stay almost unaffected by the electrostatic environment that can be realisedmore » from the charge stability diagram. Identification of different orientation and position of the Nicotine molecule within the nanopore area can be made from specific regions of overlap between different charge states on the stability diagram that could be used as an electronic fingerprint for detection. This method could be advantageous and useful to detect the presence of Nicotine in smoke which is usually performed using chemical chromatography techniques.« less
Ultrasensitive Detection of Single-Walled Carbon Nanotubes Using Surface Plasmon Resonance.
Jang, Daeho; Na, Wonhwi; Kang, Minwook; Kim, Namjoon; Shin, Sehyun
2016-01-05
Because single-walled carbon nanotubes (SWNTs) are known to be a potentially dangerous material, inducing cancers and other diseases, any possible leakage of SWNTs through an aquatic medium such as drinking water will result in a major public threat. To solve this problem, for the present study, a highly sensitive, quantitative detection method of SWNTs in an aqueous solution was developed using surface plasmon resonance (SPR) spectroscopy. For a highly sensitive and specific detection, a strong affinity conjugation with biotin-streptavidin was adopted on an SPR sensing mechanism. During the pretreatment process, the SWNT surface was functionalized and hydrophilized using a thymine-chain based biotinylated single-strand DNA linker (B-ssDNA) and bovine serum albumin (BSA). The pretreated SWNTs were captured on a sensing film, the surface of which was immobilized with streptavidin on biotinylated gold film. The captured SWNTs were measured in real-time using SPR spectroscopy. Specific binding with SWNTs was verified through several validation experiments. The present method using an SPR sensor is capable of detecting SWNTs of as low as 100 fg/mL, which is the lowest level reported thus far for carbon-nanotube detection. In addition, the SPR sensor showed a linear characteristic within the range of 100 pg/mL to 200 ng/mL. These findings imply that the present SPR sensing method can detect an extremely low level of SWNTs in an aquatic environment with high sensitivity and high specificity, and thus any potential leakage of SWNTs into an aquatic environment can be precisely monitored within a couple of hours.
Giant light-harvesting nanoantenna for single-molecule detection in ambient light
Trofymchuk, Kateryna; Reisch, Andreas; Didier, Pascal; Fras, François; Gilliot, Pierre; Mely, Yves; Klymchenko, Andrey S.
2017-01-01
Here, we explore the enhancement of single molecule emission by polymeric nano-antenna that can harvest energy from thousands of donor dyes to a single acceptor. In this nano-antenna, the cationic dyes are brought together in very close proximity using bulky counterions, thus enabling ultrafast diffusion of excitation energy (≤30 fs) with minimal losses. Our 60-nm nanoparticles containing >10,000 rhodamine-based donor dyes can efficiently transfer energy to 1-2 acceptors resulting in an antenna effect of ~1,000. Therefore, single Cy5-based acceptors become 25-fold brighter than quantum dots QD655. This unprecedented amplification of the acceptor dye emission enables observation of single molecules at illumination powers (1-10 mW cm-2) that are >10,000-fold lower than typically required in single-molecule measurements. Finally, using a basic setup, which includes a 20X air objective and a sCMOS camera, we could detect single Cy5 molecules by simply shining divergent light on the sample at powers equivalent to sunlight. PMID:28983324
Zhang, Yuqin; Lin, Fanbo; Zhang, Youyu; Li, Haitao; Zeng, Yue; Tang, Hao; Yao, Shouzhuo
2011-01-01
A new method for the detection of point mutation in DNA based on the monobase-coded cadmium tellurium nanoprobes and the quartz crystal microbalance (QCM) technique was reported. A point mutation (single-base, adenine, thymine, cytosine, and guanine, namely, A, T, C and G, mutation in DNA strand, respectively) DNA QCM sensor was fabricated by immobilizing single-base mutation DNA modified magnetic beads onto the electrode surface with an external magnetic field near the electrode. The DNA-modified magnetic beads were obtained from the biotin-avidin affinity reaction of biotinylated DNA and streptavidin-functionalized core/shell Fe(3)O(4)/Au magnetic nanoparticles, followed by a DNA hybridization reaction. Single-base coded CdTe nanoprobes (A-CdTe, T-CdTe, C-CdTe and G-CdTe, respectively) were used as the detection probes. The mutation site in DNA was distinguished by detecting the decreases of the resonance frequency of the piezoelectric quartz crystal when the coded nanoprobe was added to the test system. This proposed detection strategy for point mutation in DNA is proved to be sensitive, simple, repeatable and low-cost, consequently, it has a great potential for single nucleotide polymorphism (SNP) detection. 2011 © The Japan Society for Analytical Chemistry
Supercontinuum Fourier transform spectrometry with balanced detection on a single photodiode
DOE Office of Scientific and Technical Information (OSTI.GOV)
Goncharov, Vasily; Hall, Gregory
Here, we have developed phase-sensitive signal detection and processing algorithms for Fourier transform spectrometers fitted with supercontinuum sources for applications requiring ultimate sensitivity. Similar to well-established approach of source noise cancellation through balanced detection of monochromatic light, our method is capable of reducing the relative intensity noise of polychromatic light by 40 dB. Unlike conventional balanced detection, which relies on differential absorption measured with a well matched pair of photo-detectors, our algorithm utilizes phase-sensitive differential detection on a single photodiode and is capable of the real-time correction for instabilities in supercontinuum spectral structure over a broad range of wavelengths. Inmore » the resulting method is universal in terms of applicable wavelengths and compatible with commercial spectrometers. We present a proof-of-principle experimental« less
Supercontinuum Fourier transform spectrometry with balanced detection on a single photodiode
Goncharov, Vasily; Hall, Gregory
2016-08-25
Here, we have developed phase-sensitive signal detection and processing algorithms for Fourier transform spectrometers fitted with supercontinuum sources for applications requiring ultimate sensitivity. Similar to well-established approach of source noise cancellation through balanced detection of monochromatic light, our method is capable of reducing the relative intensity noise of polychromatic light by 40 dB. Unlike conventional balanced detection, which relies on differential absorption measured with a well matched pair of photo-detectors, our algorithm utilizes phase-sensitive differential detection on a single photodiode and is capable of the real-time correction for instabilities in supercontinuum spectral structure over a broad range of wavelengths. Inmore » the resulting method is universal in terms of applicable wavelengths and compatible with commercial spectrometers. We present a proof-of-principle experimental« less
NASA Astrophysics Data System (ADS)
Kelley, Luke Zoltan; Blecha, Laura; Hernquist, Lars; Sesana, Alberto; Taylor, Stephen R.
2018-06-01
We calculate the properties, occurrence rates and detection prospects of individually resolvable `single sources' in the low-frequency gravitational wave (GW) spectrum. Our simulations use the population of galaxies and massive black hole binaries from the Illustris cosmological hydrodynamic simulations, coupled to comprehensive semi-analytic models of the binary merger process. Using mock pulsar timing arrays (PTA) with, for the first time, varying red-noise models, we calculate plausible detection prospects for GW single sources and the stochastic GW background (GWB). Contrary to previous results, we find that single sources are at least as detectable as the GW background. Using mock PTA, we find that these `foreground' sources (also `deterministic'/`continuous') are likely to be detected with ˜20 yr total observing baselines. Detection prospects, and indeed the overall properties of single sources, are only moderately sensitive to binary evolution parameters - namely eccentricity and environmental coupling, which can lead to differences of ˜5 yr in times to detection. Red noise has a stronger effect, roughly doubling the time to detection of the foreground between a white-noise only model (˜10-15 yr) and severe red noise (˜20-30 yr). The effect of red noise on the GWB is even stronger, suggesting that single source detections may be more robust. We find that typical signal-to-noise ratios for the foreground peak near f = 0.1 yr-1, and are much less sensitive to the continued addition of new pulsars to PTA.
Single photon detection and timing in the Lunar Laser Ranging Experiment.
NASA Technical Reports Server (NTRS)
Poultney, S. K.
1972-01-01
The goals of the Lunar Laser Ranging Experiment lead to the need for the measurement of a 2.5 sec time interval to an accuracy of a nanosecond or better. The systems analysis which included practical retroreflector arrays, available laser systems, and large telescopes led to the necessity of single photon detection. Operation under all background illumination conditions required auxiliary range gates and extremely narrow spectral and spatial filters in addition to the effective gate provided by the time resolution. Nanosecond timing precision at relatively high detection efficiency was obtained using the RCA C31000F photomultiplier and Ortec 270 constant fraction of pulse-height timing discriminator. The timing accuracy over the 2.5 sec interval was obtained using a digital interval with analog vernier ends. Both precision and accuracy are currently checked internally using a triggerable, nanosecond light pulser. Future measurements using sub-nanosecond laser pulses will be limited by the time resolution of single photon detectors.
Label-free in-flow detection of single DNA molecules using glass nanopipettes.
Gong, Xiuqing; Patil, Amol V; Ivanov, Aleksandar P; Kong, Qingyuan; Gibb, Thomas; Dogan, Fatma; deMello, Andrew J; Edel, Joshua B
2014-01-07
With the view of enhancing the functionality of label-free single molecule nanopore-based detection, we have designed and developed a highly robust, mechanically stable, integrated nanopipette-microfluidic device which combines the recognized advantages of microfluidic systems and the unique properties/advantages of nanopipettes. Unlike more typical planar solid-state nanopores, which have inherent geometrical constraints, nanopipettes can be easily positioned at any point within a microfluidic channel. This is highly advantageous, especially when taking into account fluid flow properties. We show that we are able to detect and discriminate between DNA molecules of varying lengths when motivated through a microfluidic channel, upon the application of appropriate voltage bias across the nanopipette. The effects of applied voltage and volumetric flow rates have been studied to ascertain translocation event frequency and capture rate. Additionally, by exploiting the advantages associated with microfluidic systems (such as flow control and concomitant control over analyte concentration/presence), we show that the technology offers a new opportunity for single molecule detection and recognition in microfluidic devices.
Detection of isolated protein-bound metal ions by single-particle cryo-STEM.
Elad, Nadav; Bellapadrona, Giuliano; Houben, Lothar; Sagi, Irit; Elbaum, Michael
2017-10-17
Metal ions play essential roles in many aspects of biological chemistry. Detecting their presence and location in proteins and cells is important for understanding biological function. Conventional structural methods such as X-ray crystallography and cryo-transmission electron microscopy can identify metal atoms on protein only if the protein structure is solved to atomic resolution. We demonstrate here the detection of isolated atoms of Zn and Fe on ferritin, using cryogenic annular dark-field scanning transmission electron microscopy (cryo-STEM) coupled with single-particle 3D reconstructions. Zn atoms are found in a pattern that matches precisely their location at the ferroxidase sites determined earlier by X-ray crystallography. By contrast, the Fe distribution is smeared along an arc corresponding to the proposed path from the ferroxidase sites to the mineral nucleation sites along the twofold axes. In this case the single-particle reconstruction is interpreted as a probability distribution function based on the average of individual locations. These results establish conditions for detection of isolated metal atoms in the broader context of electron cryo-microscopy and tomography.
Detection of isolated protein-bound metal ions by single-particle cryo-STEM
Elad, Nadav; Bellapadrona, Giuliano; Houben, Lothar; Sagi, Irit; Elbaum, Michael
2017-01-01
Metal ions play essential roles in many aspects of biological chemistry. Detecting their presence and location in proteins and cells is important for understanding biological function. Conventional structural methods such as X-ray crystallography and cryo-transmission electron microscopy can identify metal atoms on protein only if the protein structure is solved to atomic resolution. We demonstrate here the detection of isolated atoms of Zn and Fe on ferritin, using cryogenic annular dark-field scanning transmission electron microscopy (cryo-STEM) coupled with single-particle 3D reconstructions. Zn atoms are found in a pattern that matches precisely their location at the ferroxidase sites determined earlier by X-ray crystallography. By contrast, the Fe distribution is smeared along an arc corresponding to the proposed path from the ferroxidase sites to the mineral nucleation sites along the twofold axes. In this case the single-particle reconstruction is interpreted as a probability distribution function based on the average of individual locations. These results establish conditions for detection of isolated metal atoms in the broader context of electron cryo-microscopy and tomography. PMID:28973937
Optical Detection and Sizing of Single Nano-Particles Using Continuous Wetting Films
Hennequin, Yves; McLeod, Euan; Mudanyali, Onur; Migliozzi, Daniel; Ozcan, Aydogan; Dinten, Jean-Marc
2013-01-01
The physical interaction between nano-scale objects and liquid interfaces can create unique optical properties, enhancing the signatures of the objects with sub-wavelength features. Here we show that the evaporation on a wetting substrate of a polymer solution containing sub-micrometer or nano-scale particles creates liquid micro-lenses that arise from the local deformations of the continuous wetting film. These micro-lenses have properties similar to axicon lenses that are known to create beams with a long depth of focus. This enhanced depth of focus allows detection of single nanoparticles using a low magnification microscope objective lens, achieving a relatively wide field-of-view, while also lifting the constraints on precise focusing onto the object plane. Hence, by creating these liquid axicon lenses through spatial deformations of a continuous thin wetting film, we transfer the challenge of imaging individual nano-particles to detecting the light focused by these lenses. As a proof of concept, we demonstrate the detection and sizing of single nano-particles (100 and 200 nm), CpGV granuloviruses as well as Staphylococcus epidermidis bacteria over a wide field of view of e.g., 5.10×3.75 mm2 using a ×5 objective lens with a numerical aperture of 0.15. In addition to conventional lens-based microscopy, this continuous wetting film based approach is also applicable to lensfree computational on-chip imaging, which can be used to detect single nano-particles over a large field-of-view of e.g., >20-30 mm2. These results could be especially useful for high-throughput field-analysis of nano-scale objects using compact and cost-effective microscope designs. PMID:23889001
Single-molecule DNA detection with an engineered MspA protein nanopore
Butler, Tom Z.; Pavlenok, Mikhail; Derrington, Ian M.; Niederweis, Michael; Gundlach, Jens H.
2008-01-01
Nanopores hold great promise as single-molecule analytical devices and biophysical model systems because the ionic current blockades they produce contain information about the identity, concentration, structure, and dynamics of target molecules. The porin MspA of Mycobacterium smegmatis has remarkable stability against environmental stresses and can be rationally modified based on its crystal structure. Further, MspA has a short and narrow channel constriction that is promising for DNA sequencing because it may enable improved characterization of short segments of a ssDNA molecule that is threaded through the pore. By eliminating the negative charge in the channel constriction, we designed and constructed an MspA mutant capable of electronically detecting and characterizing single molecules of ssDNA as they are electrophoretically driven through the pore. A second mutant with additional exchanges of negatively-charged residues for positively-charged residues in the vestibule region exhibited a factor of ≈20 higher interaction rates, required only half as much voltage to observe interaction, and allowed ssDNA to reside in the vestibule ≈100 times longer than the first mutant. Our results introduce MspA as a nanopore for nucleic acid analysis and highlight its potential as an engineerable platform for single-molecule detection and characterization applications. PMID:19098105
Elastic light single-scattering spectroscopy for detection of dysplastic tissues
NASA Astrophysics Data System (ADS)
Canpolat, Murat; Denkçeken, Tuba; Akman, Ayşe.; Alpsoy, Erkan; Tuncer, Recai; Akyüz, Mahmut; Baykara, Mehmet; Yücel, Selçuk; Başsorgun, Ibrahim; ćiftçioǧlu, M. Akif; Gökhan, Güzide Ayşe.; Gürer, ElifInanç; Peştereli, Elif; Karaveli, Šeyda
2013-11-01
Elastic light single-scattering spectroscopy (ELSSS) system has been developed and tested in diagnosis of cancerous tissues of different organs. ELSSS system consists of a miniature visible light spectrometer, a single fiber optical probe, a halogen tungsten light source and a laptop. Measurements were performed on excised brain, skin, cervix and prostate tumor specimens and surrounding normal tissues. Single fiber optical probe with a core diameter of 100 μm was used to deliver white light to and from tissue. Single optical fiber probe mostly detects singly scattered light from tissue rather than diffused light. Therefore, measured spectra are sensitive to size of scatters in tissue such as cells, nuclei, mitochondria and other organelles of cells. Usually, nuclei of tumor cells are larger than nuclei of normal cells. Therefore, spectrum of singly scattered light of tumor tissue is different than normal tissue. The spectral slopes were shown to be positive for normal brain, skin and prostate and cervix tissues and negative for the tumors of the same tissues. Signs of the spectral slopes were used as a discrimination parameter to differentiate tumor from normal tissues for the three organ tissues. Sensitivity and specificity of the system in differentiation between tumors from normal tissues were 93% and %100 for brain, 87% and 85% for skin, 93.7% and 46.1% for cervix and 98% and 100% for prostate.
Multimodal autofluorescence detection of cancer: from single cells to living organism
NASA Astrophysics Data System (ADS)
Horilova, J.; Cunderlikova, B.; Cagalinec, M.; Chorvat, D.; Marcek Chorvatova, A.
2018-02-01
Multimodal optical imaging of suspected tissues is showing to be a promising method for distinguishing suspected cancerous tissues from healthy ones. In particular, the combination of steady-state spectroscopic methods with timeresolved fluorescence provides more precise insight into native metabolism when focused on tissue autofluorescence. Cancer is linked to specific metabolic remodelation detectable spectroscopically. In this work, we evaluate possibilities and limitations of multimodal optical cancer detection in single cells, collagen-based 3D cell cultures and in living organisms (whole mice), as a representation of gradually increasing complexity of model systems.
Single-image-based Rain Detection and Removal via CNN
NASA Astrophysics Data System (ADS)
Chen, Tianyi; Fu, Chengzhou
2018-04-01
The quality of the image is degraded by rain streaks, which have negative impact when we extract image features for many visual tasks, such as feature extraction for classification and recognition, tracking, surveillance and autonomous navigation. Hence, it is necessary to detect and remove rain streaks from single images, which is a challenging problem since we have no spatial-temporal information of rain streaks compared to the dynamic video stream. Inspired by the priori that the rain streaks have almost the same feature, such as the direction or the thickness, although they are in different types of real-world images. The paper aims at proposing an effective convolutional neural network (CNN) to detect and remove rain streaks from single image. Two models of synthesized rainy image, linear additive composite model (LACM model) and screen blend model (SCM model), are considered in this paper. The main idea is that it is easier for our CNN network to find the mapping between the rainy image and rain streaks than between the rainy image and clean image. The reason is that rain streaks have fixed features, but clean images have various features. The experiments show that the designed CNN network outperforms state-of-the-art approaches on both synthesized and real-world images, which indicates the effectiveness of our proposed framework.
Wang, Yuling; Rauf, Sakandar; Grewal, Yadveer S; Spadafora, Lauren J; Shiddiky, Muhammad J A; Cangelosi, Gerard A; Schlücker, Sebastian; Trau, Matt
2014-10-07
Quantitative and accurate detection of multiple biomarkers would allow for the rapid diagnosis and treatment of diseases induced by pathogens. Monoclonal antibodies are standard affinity reagents applied for biomarkers detection; however, their production is expensive and labor-intensive. Herein, we report on newly developed nanoyeast single-chain variable fragments (NYscFv) as an attractive alternative to monoclonal antibodies, which offers the unique advantage of a cost-effective production, stability in solution, and target-specificity. By combination of surface-enhanced Raman scattering (SERS) microspectroscopy using glass-coated, highly purified SERS nanoparticle clusters as labels, with a microfluidic device comprising multiple channels, a robust platform for the sensitive duplex detection of pathogen antigens has been developed. Highly sensitive detection for individual Entamoeba histolytica antigen EHI_115350 (limit of detection = 1 pg/mL, corresponding to 58.8 fM) and EHI_182030 (10 pg/mL, corresponding 453 fM) with high specificity has been achieved, employing the newly developed corresponding NYscFv as probe in combination with SERS microspectroscopy at a single laser excitation wavelength. Our first report on SERS-based immunoassays using the novel NYscFv affinity reagent demonstrates the flexibility of NYscFv fragments as viable alternatives to monoclonal antibodies in a range of bioassay platforms and paves the way for further applications.
Martin, Audrey N; Farquar, George R; Frank, Matthias; Gard, Eric E; Fergenson, David P
2007-08-15
Single-particle aerosol mass spectrometry (SPAMS) was used for the real-time detection of liquid nerve agent simulants. A total of 1000 dual-polarity time-of-flight mass spectra were obtained for micrometer-sized single particles each of dimethyl methyl phosphonate, diethyl ethyl phosphonate, diethyl phosphoramidate, and diethyl phthalate using laser fluences between 0.58 and 7.83 nJ/microm2, and mass spectral variation with laser fluence was studied. The mass spectra obtained allowed identification of single particles of the chemical warfare agent (CWA) simulants at each laser fluence used although lower laser fluences allowed more facile identification. SPAMS is presented as a promising real-time detection system for the presence of CWAs.
Single virus particle mass detection using microresonators with nanoscale thickness
NASA Astrophysics Data System (ADS)
Gupta, A.; Akin, D.; Bashir, R.
2004-03-01
In this letter, we present the microfabrication and application of arrays of silicon cantilever beams as microresonator sensors with nanoscale thickness to detect the mass of individual virus particles. The dimensions of the fabricated cantilever beams were in the range of 4-5 μm in length, 1-2 μm in width and 20-30 nm in thickness. The virus particles we used in the study were vaccinia virus, which is a member of the Poxviridae family and forms the basis of the smallpox vaccine. The frequency spectra of the cantilever beams, due to thermal and ambient noise, were measured using a laser Doppler vibrometer under ambient conditions. The change in resonant frequency as a function of the virus particle mass binding on the cantilever beam surface forms the basis of the detection scheme. We have demonstrated the detection of a single vaccinia virus particle with an average mass of 9.5 fg. These devices can be very useful as components of biosensors for the detection of airborne virus particles.
Kawakami, Shuji; Hasegawa, Takuya; Imachi, Hiroyuki; Yamaguchi, Takashi; Harada, Hideki; Ohashi, Akiyoshi; Kubota, Kengo
2012-02-01
In situ detection of functional genes with single-cell resolution is currently of interest to microbiologists. Here, we developed a two-pass tyramide signal amplification (TSA)-fluorescence in situ hybridization (FISH) protocol with PCR-derived polynucleotide probes for the detection of single-copy genes in prokaryotic cells. The mcrA gene and the apsA gene in methanogens and sulfate-reducing bacteria, respectively, were targeted. The protocol showed bright fluorescence with a good signal-to-noise ratio and achieved a high efficiency of detection (>98%). The discrimination threshold was approximately 82-89% sequence identity. Microorganisms possessing the mcrA or apsA gene in anaerobic sludge samples were successfully detected by two-pass TSA-FISH with polynucleotide probes. The developed protocol is useful for identifying single microbial cells based on functional gene sequences. Copyright © 2011 Elsevier B.V. All rights reserved.
Single-pixel imaging using balanced detection and a digital micromirror device
NASA Astrophysics Data System (ADS)
Soldevila, F.; Clemente, P.; Tajahuerce, E.; Uribe-Patarroyo, Néstor; Andrés, P.; Lancis, J.
2018-02-01
Over the past decade, single-pixel imaging (SPI) has established as a viable tool in scenarios where traditional imaging techniques struggle to provide images with acceptable quality in practicable times and reasonable costs. However, SPI still has several limitations inherent to the technique, such as working with spurious light and in real time. Here we present a novel approach, using complementary measurements and a single balanced detector. By using balanced detection, we improve the frame rate of the complementary measurement architectures by a factor of two. Furthermore, the use of a balanced detector provides environmental light immunity to the method.
Mapping yeast origins of replication via single-stranded DNA detection.
Feng, Wenyi; Raghuraman, M K; Brewer, Bonita J
2007-02-01
Studies in th Saccharomyces cerevisiae have provided a framework for understanding how eukaryotic cells replicate their chromosomal DNA to ensure faithful transmission of genetic information to their daughter cells. In particular, S. cerevisiae is the first eukaryote to have its origins of replication mapped on a genomic scale, by three independent groups using three different microarray-based approaches. Here we describe a new technique of origin mapping via detection of single-stranded DNA in yeast. This method not only identified the majority of previously discovered origins, but also detected new ones. We have also shown that this technique can identify origins in Schizosaccharomyces pombe, illustrating the utility of this method for origin mapping in other eukaryotes.
Decrop, Deborah; Ruiz, Elena Pérez; Kumar, Phalguni Tewari; Tripodi, Lisa; Kokalj, Tadej; Lammertyn, Jeroen
2017-01-01
Digital microfluidics has emerged in the last years as a promising liquid handling technology for a variety of applications. Here, we describe in detail how to build up an electrowetting-on-dielectric-based digital microfluidic chip with unique advantages for performing single-molecule detection. We illustrate how superparamagnetic particles can be printed with very high loading efficiency (over 98 %) and single-particle resolution in the microwell array patterned in the Teflon-AF ® surface of the grounding plate of the chip. Finally, the potential of the device for its application to single-molecule detection is demonstrated by the ultrasensitive detection of the biotinylated enzyme β-Galactosidase captured on streptavidin-coated particles in the described platform.
Low contrast detection in abdominal CT: comparing single-slice and multi-slice tasks
NASA Astrophysics Data System (ADS)
Ba, Alexandre; Racine, Damien; Viry, Anaïs.; Verdun, Francis R.; Schmidt, Sabine; Bochud, François O.
2017-03-01
Image quality assessment is crucial for the optimization of computed tomography (CT) protocols. Human and mathematical model observers are increasingly used for the detection of low contrast signal in abdominal CT, but are frequently limited to the use of a single image slice. Another limitation is that most of them only consider the detection of a signal embedded in a uniform background phantom. The purpose of this paper was to test if human observer performance is significantly different in CT images read in single or multiple slice modes and if these differences are the same for anatomical and uniform clinical images. We investigated detection performance and scrolling trends of human observers of a simulated liver lesion embedded in anatomical and uniform CT backgrounds. Results show that observers don't take significantly benefit of additional information provided in multi-slice reading mode. Regarding the background, performances are moderately higher for uniform than for anatomical images. Our results suggest that for low contrast detection in abdominal CT, the use of multi-slice model observers would probably only add a marginal benefit. On the other hand, the quality of a CT image is more accurately estimated with clinical anatomical backgrounds.
Supercontinuum Fourier transform spectrometry with balanced detection on a single photodiode
DOE Office of Scientific and Technical Information (OSTI.GOV)
Goncharov, Vasily V.; Hall, Gregory E., E-mail: gehall@bnl.gov
We demonstrate a method of combining a supercontinuum light source with a commercial Fourier transform spectrometer, using a novel approach to dual-beam balanced detection, implemented with phase-sensitive detection on a single light detector. A 40 dB reduction in the relative intensity noise is achieved for broadband light, analogous to conventional balanced detection methods using two matched photodetectors. Unlike conventional balanced detection, however, this method exploits the time structure of the broadband source to interleave signal and reference pulse trains in the time domain, recording the broadband differential signal at the fundamental pulse repetition frequency of the supercontinuum. The method ismore » capable of real-time correction for instability in the supercontinuum spectral structure over a broad range of wavelengths and is compatible with commercially designed spectrometers. A proof-of-principle experimental setup is demonstrated for weak absorption in the 1500-1600 nm region.« less
Hellmich, Wibke; Greif, Dominik; Pelargus, Christoph; Anselmetti, Dario; Ros, Alexandra
2006-10-20
Single cell analytics is a key method in the framework of proteom research allowing analyses, which are not subjected to ensemble-averaging, cell-cycle or heterogeneous cell-population effects. Our previous studies on single cell analysis in poly(dimethylsiloxane) microfluidic devices with native label-free laser induced fluorescence detection [W. Hellmich, C. Pelargus, K. Leffhalm, A. Ros, D. Anselmetti, Electrophoresis 26 (2005) 3689] were extended in order to improve separation efficiency and detection sensitivity. Here, we particularly focus on the influence of poly(oxyethylene) based coatings on the separation performance. In addition, the influence on background fluorescence is studied by the variation of the incident laser power as well as the adaptation of the confocal volume to the microfluidic channel dimensions. Last but not least, the use of carbon black particles further enhanced the detection limit to 25 nM, thereby reaching the relevant concentration ranges necessary for the label-free detection of low abundant proteins in single cells. On the basis of these results, we demonstrate the first electropherogram from an individual Spodoptera frugiperda (Sf9) cell with native label-free UV-LIF detection in a microfluidic chip.
Tu, Yiheng; Hung, Yeung Sam; Hu, Li; Huang, Gan; Hu, Yong; Zhang, Zhiguo
2014-12-01
This study aims (1) to develop an automated and fast approach for detecting visual evoked potentials (VEPs) in single trials and (2) to apply the single-trial VEP detection approach in designing a real-time and high-performance brain-computer interface (BCI) system. The single-trial VEP detection approach uses common spatial pattern (CSP) as a spatial filter and wavelet filtering (WF) a temporal-spectral filter to jointly enhance the signal-to-noise ratio (SNR) of single-trial VEPs. The performance of the joint spatial-temporal-spectral filtering approach was assessed in a four-command VEP-based BCI system. The offline classification accuracy of the BCI system was significantly improved from 67.6±12.5% (raw data) to 97.3±2.1% (data filtered by CSP and WF). The proposed approach was successfully implemented in an online BCI system, where subjects could make 20 decisions in one minute with classification accuracy of 90%. The proposed single-trial detection approach is able to obtain robust and reliable VEP waveform in an automatic and fast way and it is applicable in VEP based online BCI systems. This approach provides a real-time and automated solution for single-trial detection of evoked potentials or event-related potentials (EPs/ERPs) in various paradigms, which could benefit many applications such as BCI and intraoperative monitoring. Copyright © 2014 International Federation of Clinical Neurophysiology. Published by Elsevier Ireland Ltd. All rights reserved.
NASA Astrophysics Data System (ADS)
Marsden, Nicholas; Flynn, Michael J.; Taylor, Jonathan W.; Allan, James D.; Coe, Hugh
2016-12-01
Single-particle mass spectrometry (SPMS) is a useful tool for the online study of aerosols with the ability to measure size-resolved chemical composition with a temporal resolution relevant to atmospheric processes. In SPMS, optical particle detection is used for the effective temporal alignment of an ablation laser pulse with the presence of a particle in the ion source, and it gives the option of aerodynamic sizing by measuring the offset of particle arrival times between two detection stages. The efficiency of the optical detection stage has a strong influence on the overall instrument performance. A custom detection laser system consisting of a high-powered fibre-coupled Nd:YAG solid-state laser with a collimated beam was implemented in the detection stage of a laser ablation aerosol particle time-of-flight (LAAP-TOF) single-particle mass spectrometer without major modifications to instrument geometry. The use of a collimated laser beam permitted the construction of a numerical model that predicts the effects of detection laser wavelength, output power, beam focussing characteristics, light collection angle, particle size, and refractive index on the effective detection radius (R) of the detection laser beam. We compare the model predictions with an ambient data set acquired during the Ice in Clouds Experiment - Dust (ICE-D) project. The new laser system resulted in an order-of-magnitude improvement in instrument sensitivity to spherical particles in the size range 500-800 nm compared to a focussed 405 nm laser diode system. The model demonstrates that the limit of detection in terms of particle size is determined by the scattering cross section (Csca) as predicted by Mie theory. In addition, if light is collected over a narrow collection angle, oscillations in the magnitude of Csca with respect to particle diameter result in a variation in R, resulting in large particle-size-dependent variation in detection efficiency across the particle transmission range
McCutchen-Maloney, Sandra L.
2002-01-01
DNA mutation binding proteins alone and as chimeric proteins with nucleases are used with solid supports to detect DNA sequence variations, DNA mutations and single nucleotide polymorphisms. The solid supports may be flow cytometry beads, DNA chips, glass slides or DNA dips sticks. DNA molecules are coupled to solid supports to form DNA-support complexes. Labeled DNA is used with unlabeled DNA mutation binding proteins such at TthMutS to detect DNA sequence variations, DNA mutations and single nucleotide length polymorphisms by binding which gives an increase in signal. Unlabeled DNA is utilized with labeled chimeras to detect DNA sequence variations, DNA mutations and single nucleotide length polymorphisms by nuclease activity of the chimera which gives a decrease in signal.
Detection of gastritis by single- and double-contrast radiography
DOE Office of Scientific and Technical Information (OSTI.GOV)
Thoeni, R.F.; Goldberg, H.I.; Ominsky, S.
1983-09-01
Sixty-eight patients with various types of gastritis, 23 patients with normal stomachs, and four patients with other gastric diseases were examined in a prospective study to assess the sensitivity and specificity of single-contrast (SC) and double-contrast (DC) upper gastrointestinal examinations in the evaluation of gastritis. All patients underwent endoscopy with biopsy followed first by DC and then by SC radiography. The respective sensitivities of SC and DC radiography were 58% and 72% for all examinations and 59% and 77% for adequate examinations only. The respective specificities were 59% and 55% based on all examinations. Useful radiographic features included polypoid defectsmore » and erosions detected by both methods, abnormal folds and flattened margins detected by the SC technique, and narrowed lumen and crenulated margins detected by the DC technique. In 93% of all cases, the correct diagnosis was based on two or more of these radiographic features. According to this study, the radiographic sensitivity in the detection of gastritis is reliable only in cases of moderate-to-severe disease and only when based on findings of the DC examination. Neither SC nor DC radiography should be used as the primary screening method for patients with suspected gastritis, and the radiographic diagnosis should be restricted to the terms ''erosive'' or ''nonerosive gastritis.''« less
Development of Single-Stranded DNA Aptamers for Specific Bisphenol A Detection
Jo, Minjoung; Ahn, Ji-Young; Lee, Joohyung; Lee, Seram; Hong, Sun Woo; Yoo, Jae-Wook; Kang, Jeehye; Dua, Pooja
2011-01-01
The development of reagents with high affinity and specificity to small molecules is crucial for the high-throughput detection of chemical compounds, such as toxicants or pollutants. Aptamers are short and single-stranded (ss) oligonucleotides able to recognize target molecules with high affinity. Here, we report the selection of ssDNA aptamers that bind to Bisphenol A (BPA), an environmental hormone. Using SELEX process, we isolated high affinity aptamers to BPA from a 1015 random library of 60 mer ssDNAs. The selected aptamers bound specifically to BPA, but not to structurally similar molecules, such as Bisphenol B with one methyl group difference, or 4,4′-Bisphenol with 2 methyl groups difference. Using these aptamers, we developed an aptamer-based sol–gel biochip and detected BPA dissolved in water. This novel BPA aptamer-based detection can be further applied to the universal and high-specificity detection of small molecules. PMID:21413891
Metallic single-walled carbon nanotube for ionized radiation detection
NASA Astrophysics Data System (ADS)
Banadaki, Yaser M.; Srivastava, Ashok; Sharifi, Safura
2016-04-01
In this paper, we have explored the feasibility of a metallic single-walled carbon nanotube (SWCNT) as a radiation detector. The effect of SWCNTs' exposure to different ion irradiations is considered with the displacement damage dose (DDD) methodology. The analytical model of the irradiated resistance of metallic SWCNT has been developed and verified by the experimental data for increasing DDD from 1012 MeV/g to 1017 MeV/g. It has been found that the resistance variation of SWCNT by increasing DDD can be significant depending on the length and diameter of SWCNT, such that the DDD as low as 1012 (MeV/g) can be detected using the SWCNT with 1cm length and 5nm diameter. Increasing the length and diameter of SWCNT can result in both the higher radiation sensitivity of resistance and the extension of detection range to lower DDD.
Single ricin detection by atomic force microscopy chemomechanical mapping
NASA Astrophysics Data System (ADS)
Chen, Guojun; Zhou, Jianfeng; Park, Bosoon; Xu, Bingqian
2009-07-01
The authors report on a study of detecting ricin molecules immobilized on chemically modified Au (111) surface by chemomechanically mapping the molecular interactions with a chemically modified atomic force microscopy (AFM) tip. AFM images resolved the different fold-up conformations of single ricin molecule as well as their intramolecule structure of A- and B-chains. AFM force spectroscopy study of the interaction indicates that the unbinding force has a linear relation with the logarithmic force loading rate, which agrees well with calculations using one-barrier bond dissociation model.
Single-Particle Detection of Transcription following Rotavirus Entry
Salgado, Eric N.; Upadhyayula, Srigokul
2017-01-01
ABSTRACT Infectious rotavirus particles are triple-layered, icosahedral assemblies. The outer layer proteins, VP4 (cleaved to VP8* and VP5*) and VP7, surround a transcriptionally competent, double-layer particle (DLP), which they deliver into the cytosol. During entry of rhesus rotavirus, VP8* interacts with cell surface gangliosides, allowing engulfment into a membrane vesicle by a clathrin-independent process. Escape into the cytosol and outer-layer shedding depend on interaction of a hydrophobic surface on VP5* with the membrane bilayer and on a large-scale conformational change. We report here experiments that detect the fate of released DLPs and their efficiency in initiating RNA synthesis. By replacing the outer layer with fluorescently tagged, recombinant proteins and also tagging the DLP, we distinguished particles that have lost their outer layer and entered the cytosol (uncoated) from those still within membrane vesicles. We used fluorescent in situ hybridization with probes for nascent transcripts to determine how soon after uncoating transcription began and what fraction of the uncoated particles were active in initiating RNA synthesis. We detected RNA synthesis by uncoated particles as early as 15 min after adding virus. The uncoating efficiency was 20 to 50%; of the uncoated particles, about 10 to 15% synthesized detectable RNA. In the format of our experiments, about 10% of the added particles attached to the cell surface, giving an overall ratio of added particles to RNA-synthesizing particles of between 250:1 and 500:1, in good agreement with the ratio of particles to focus-forming units determined by infectivity assays. Thus, RNA synthesis by even a single, uncoated particle can initiate infection in a cell. IMPORTANCE The pathways by which a virus enters a cell transform its packaged genome into an active one. Contemporary fluorescence microscopy can detect individual virus particles as they enter cells, allowing us to map their multistep entry
In-flight automatic detection of vigilance states using a single EEG channel.
Sauvet, F; Bougard, C; Coroenne, M; Lely, L; Van Beers, P; Elbaz, M; Guillard, M; Leger, D; Chennaoui, M
2014-12-01
Sleepiness and fatigue can reach particularly high levels during long-haul overnight flights. Under these conditions, voluntary or even involuntary sleep periods may occur, increasing the risk of accidents. The aim of this study was to assess the performance of an in-flight automatic detection system of low-vigilance states using a single electroencephalogram channel. Fourteen healthy pilots voluntarily wore a miniaturized brain electrical activity recording device during long-haul flights ( 10 ±2.0 h, Atlantic 2 and Falcon 50 M, French naval aviation). No subject was disturbed by the equipment. Seven pilots experienced at least a period of voluntary ( 26.8 ±8.0 min, n = 4) or involuntary sleep (N1 sleep stage, 26.6 ±18.7 s, n = 7) during the flight. Automatic classification (wake/sleep) by the algorithm was made for 10-s epochs (O1-M2 or C3-M2 channel), based on comparison of means to detect changes in α, β, and θ relative power, or ratio [( α+θ)/β], or fuzzy logic fusion (α, β). Pertinence and prognostic of the algorithm were determined using epoch-by-epoch comparison with visual-scoring (two blinded readers, AASM rules). The best concordance between automatic detection and visual-scoring was observed within the O1-M2 channel, using the ratio [( α+θ )/β] ( 98.3 ±4.1% of good detection, K = 0.94 ±0.07, with a 0.04 ±0.04 false positive rate and a 0.87 ±0.10 true positive rate). Our results confirm the efficiency of a miniaturized single electroencephalographic channel recording device, associated with an automatic detection algorithm, in order to detect low-vigilance states during real flights.
Performance evaluation of an automated single-channel sleep–wake detection algorithm
Kaplan, Richard F; Wang, Ying; Loparo, Kenneth A; Kelly, Monica R; Bootzin, Richard R
2014-01-01
Background A need exists, from both a clinical and a research standpoint, for objective sleep measurement systems that are both easy to use and can accurately assess sleep and wake. This study evaluates the output of an automated sleep–wake detection algorithm (Z-ALG) used in the Zmachine (a portable, single-channel, electroencephalographic [EEG] acquisition and analysis system) against laboratory polysomnography (PSG) using a consensus of expert visual scorers. Methods Overnight laboratory PSG studies from 99 subjects (52 females/47 males, 18–60 years, median age 32.7 years), including both normal sleepers and those with a variety of sleep disorders, were assessed. PSG data obtained from the differential mastoids (A1–A2) were assessed by Z-ALG, which determines sleep versus wake every 30 seconds using low-frequency, intermediate-frequency, and high-frequency and time domain EEG features. PSG data were independently scored by two to four certified PSG technologists, using standard Rechtschaffen and Kales guidelines, and these score files were combined on an epoch-by-epoch basis, using a majority voting rule, to generate a single score file per subject to compare against the Z-ALG output. Both epoch-by-epoch and standard sleep indices (eg, total sleep time, sleep efficiency, latency to persistent sleep, and wake after sleep onset) were compared between the Z-ALG output and the technologist consensus score files. Results Overall, the sensitivity and specificity for detecting sleep using the Z-ALG as compared to the technologist consensus are 95.5% and 92.5%, respectively, across all subjects, and the positive predictive value and the negative predictive value for detecting sleep are 98.0% and 84.2%, respectively. Overall κ agreement is 0.85 (approaching the level of agreement observed among sleep technologists). These results persist when the sleep disorder subgroups are analyzed separately. Conclusion This study demonstrates that the Z-ALG automated sleep
Single-electron detection and spectroscopy via relativistic cyclotron radiation
Asner, D. M.; Bradley, R. F.; de Viveiros, L.; ...
2015-04-20
Since 1897, we've understood that accelerating charges must emit electromagnetic radiation. Cyclotron radiation, the particular form of radiation emitted by an electron orbiting in a magnetic field, was first derived in 1904. Despite the simplicity of this concept, and the enormous utility of electron spectroscopy in nuclear and particle physics, single-electron cyclotron radiation has never been observed directly. We demonstrate single-electron detection in a novel radiofrequency spec- trometer. Here, we observe the cyclotron radiation emitted by individual magnetically-trapped electrons that are produced with mildly-relativistic energies by a gaseous radioactive source. The relativistic shift in the cyclotron frequency permits a precisemore » electron energy measurement. Precise beta electron spectroscopy from gaseous radiation sources is a key technique in modern efforts to measure the neutrino mass via the tritium decay endpoint, and this work demonstrates a fundamentally new approach to precision beta spectroscopy for future neutrino mass experiments.« less
Detection of Activating Estrogen Receptor Gene (ESR1) Mutations in Single Circulating Tumor Cells.
Paolillo, Carmela; Mu, Zhaomei; Rossi, Giovanna; Schiewer, Matthew J; Nguyen, Thomas; Austin, Laura; Capoluongo, Ettore; Knudsen, Karen; Cristofanilli, Massimo; Fortina, Paolo
2017-10-15
Purpose: Early detection is essential for treatment plans before onset of metastatic disease. Our purpose was to demonstrate feasibility to detect and monitor estrogen receptor 1 ( ESR1 ) gene mutations at the single circulating tumor cell (CTC) level in metastatic breast cancer (MBC). Experimental Design: We used a CTC molecular characterization approach to investigate heterogeneity of 14 hotspot mutations in ESR1 and their correlation with endocrine resistance. Combining the CellSearch and DEPArray technologies allowed recovery of 71 single CTCs and 12 WBC from 3 ER-positive MBC patients. Forty CTCs and 12 WBC were subjected to whole genome amplification by MALBAC and Sanger sequencing. Results: Among 3 selected patients, 2 had an ESR1 mutation (Y537). One showed two different ESR1 variants in a single CTC and another showed loss of heterozygosity. All mutations were detected in matched cell-free DNA (cfDNA). Furthermore, one had 2 serial blood samples analyzed and showed changes in both cfDNA and CTCs with emergence of mutations in ESR1 (Y537S and T570I), which has not been reported previously. Conclusions: CTCs are easily accessible biomarkers to monitor and better personalize management of patients with previously demonstrated ER-MBC who are progressing on endocrine therapy. We showed that single CTC analysis can yield important information on clonal heterogeneity and can be a source of discovery of novel and potential driver mutations. Finally, we also validate a workflow for liquid biopsy that will facilitate early detection of ESR1 mutations, the emergence of endocrine resistance and the choice of further target therapy. Clin Cancer Res; 23(20); 6086-93. ©2017 AACR . ©2017 American Association for Cancer Research.
Authenticated Quantum Key Distribution with Collective Detection using Single Photons
NASA Astrophysics Data System (ADS)
Huang, Wei; Xu, Bing-Jie; Duan, Ji-Tong; Liu, Bin; Su, Qi; He, Yuan-Hang; Jia, Heng-Yue
2016-10-01
We present two authenticated quantum key distribution (AQKD) protocols by utilizing the idea of collective (eavesdropping) detection. One is a two-party AQKD protocol, the other is a multiparty AQKD protocol with star network topology. In these protocols, the classical channels need not be assumed to be authenticated and the single photons are used as the quantum information carriers. To achieve mutual identity authentication and establish a random key in each of the proposed protocols, only one participant should be capable of preparing and measuring single photons, and the main quantum ability that the rest of the participants should have is just performing certain unitary operations. Security analysis shows that these protocols are free from various kinds of attacks, especially the impersonation attack and the man-in-the-middle (MITM) attack.
Chahin, Nassif; Uribe, Laura A; Debela, Ahmed M; Thorimbert, Serge; Hasenknopf, Bernold; Ortiz, Mayreli; Katakis, Ioannis; O'Sullivan, Ciara K
2018-06-07
Polyoxymetalates (POMs) ([SiW 11 O 39 {Sn(CH 2 ) 2 CO)}] 4- and [P 2 W 17 O 61 {Sn(CH 2 ) 2 CO)}] 6- ) were used to modify dideoxynucleotides (ddNTPs) through amide bond formation, and applied to the multiplexed detection of single nucleotide polymorphisms (SNPs) in an electrochemical primer extension reaction. Each gold electrode of an array was functionalised with a short single stranded thiolated DNA probe, specifically designed to extend with the POM-ddNTP at the SNP site to be interrogated. The system was applied to the simultaneous detection of 4 SNPs within a single stranded 103-mer model target generated using asymmetric PCR, highlighting the potential of POM-ddNTPs for targeted, multiplexed SNP detection. The four DNA bases were successfully labelled with both ([SiW 11 O 39 {Sn(CH 2 ) 2 CO)}] 4- and [P 2 W 17 O 61 {Sn(CH 2 ) 2 CO)}] 6- ), and [SiW 11 O 39 {Sn(CH 2 ) 2 CO)}] 4- demonstrated to be the more suitable due to its single oxidation peak, which provides an unequivocal signal. The POM-ddNTP enzymatically incorporated to the DNA anchored to the surface was visualised by AFM using gold coated mica. The developed assay has been demonstrated to be highly reproducible, simple to carry out and with very low non-specific background signals. Future work will focus on applying the developed platform to the detection of SNPs associated with rifampicin resistance in real samples from patients suffering from tuberculosis. Copyright © 2018. Published by Elsevier B.V.
Open-source meteor detection software for low-cost single-board computers
NASA Astrophysics Data System (ADS)
Vida, D.; Zubović, D.; Šegon, D.; Gural, P.; Cupec, R.
2016-01-01
This work aims to overcome the current price threshold of meteor stations which can sometimes deter meteor enthusiasts from owning one. In recent years small card-sized computers became widely available and are used for numerous applications. To utilize such computers for meteor work, software which can run on them is needed. In this paper we present a detailed description of newly-developed open-source software for fireball and meteor detection optimized for running on low-cost single board computers. Furthermore, an update on the development of automated open-source software which will handle video capture, fireball and meteor detection, astrometry and photometry is given.
Yang, Liu; Wang, Zhihua; Deng, Yuliang; Li, Yan; Wei, Wei; Shi, Qihui
2016-11-15
Circulating tumor cells (CTCs) shed from tumor sites and represent the molecular characteristics of the tumor. Besides genetic and transcriptional characterization, it is important to profile a panel of proteins with single-cell precision for resolving CTCs' phenotype, organ-of-origin, and drug targets. We describe a new technology that enables profiling multiple protein markers of extraordinarily rare tumor cells at the single-cell level. This technology integrates a microchip consisting of 15000 60 pL-sized microwells and a novel beads-on-barcode antibody microarray (BOBarray). The BOBarray allows for multiplexed protein detection by assigning two independent identifiers (bead size and fluorescent color) of the beads to each protein. Four bead sizes (1.75, 3, 4.5, and 6 μm) and three colors (blue, green, and yellow) are utilized to encode up to 12 different proteins. The miniaturized BOBarray can fit an array of 60 pL-sized microwells that isolate single cells for cell lysis and the subsequent detection of protein markers. An enclosed 60 pL-sized microchamber defines a high concentration of proteins released from lysed single cells, leading to single-cell resolution of protein detection. The protein markers assayed in this study include organ-specific markers and drug targets that help to characterize the organ-of-origin and drug targets of isolated rare tumor cells from blood samples. This new approach enables handling a very small number of cells and achieves single-cell, multiplexed protein detection without loss of rare but clinically important tumor cells.
Detecting Bot-Answerable Questions in Ubuntu Chat
2013-10-01
another user’s question (see 1http://ubottu.com/ [13:19] <p5yx> is the netbook remix not available anymore? [13:20] <histo> !unr | p5yx [13:20...ubottu> p5yx: Starting with Ubuntu 11.04, the Ubuntu Netbook Edition is no longer being offered as a separate install as Unity is now standard for all
RNA-templated single-base mutation detection based on T4 DNA ligase and reverse molecular beacon.
Tang, Hongxing; Yang, Xiaohai; Wang, Kemin; Tan, Weihong; Li, Huimin; He, Lifang; Liu, Bin
2008-06-15
A novel RNA-templated single-base mutation detection method based on T4 DNA ligase and reverse molecular beacon (rMB) has been developed and successfully applied to identification of single-base mutation in codon 273 of the p53 gene. The discrimination was carried out using allele-specific primers, which flanked the variable position in the target RNA and was ligated using T4 DNA ligase only when the primers perfectly matched the RNA template. The allele-specific primers also carried complementary stem structures with end-labels (fluorophore TAMRA, quencher DABCYL), which formed a molecular beacon after RNase H digestion. One-base mismatch can be discriminated by analyzing the change of fluorescence intensity before and after RNase H digestion. This method has several advantages for practical applications, such as direct discrimination of single-base mismatch of the RNA extracted from cell; no requirement of PCR amplification; performance of homogeneous detection; and easily design of detection probes.
Using the time shift in single pushbroom datatakes to detect ships and their heading
NASA Astrophysics Data System (ADS)
Willburger, Katharina A. M.; Schwenk, Kurt
2017-10-01
The detection of ships from remote sensing data has become an essential task for maritime security. The variety of application scenarios includes piracy, illegal fishery, ocean dumping and ships carrying refugees. While techniques using data from SAR sensors for ship detection are widely common, there is only few literature discussing algorithms based on imagery of optical camera systems. A ship detection algorithm for optical pushbroom data has been developed. It takes advantage of the special detector assembly of most of those scanners, which allows apart from the detection of a ship also the calculation of its heading out of a single acquisition. The proposed algorithm for the detection of moving ships was developed with RapidEye imagery. It algorithm consists mainly of three steps: the creation of a land-watermask, the object extraction and the deeper examination of each single object. The latter step is built up by several spectral and geometric filters, making heavy use of the inter-channel displacement typical for pushbroom sensors with multiple CCD lines, finally yielding a set of ships and their direction of movement. The working principle of time-shifted pushbroom sensors and the developed algorithm is explained in detail. Furthermore, we present our first results and give an outlook to future improvements.
Single-shot detection of bacterial endospores via coherent Raman spectroscopy.
Pestov, Dmitry; Wang, Xi; Ariunbold, Gombojav O; Murawski, Robert K; Sautenkov, Vladimir A; Dogariu, Arthur; Sokolov, Alexei V; Scully, Marlan O
2008-01-15
Recent advances in coherent Raman spectroscopy hold exciting promise for many potential applications. For example, a technique, mitigating the nonresonant four-wave-mixing noise while maximizing the Raman-resonant signal, has been developed and applied to the problem of real-time detection of bacterial endospores. After a brief review of the technique essentials, we show how extensions of our earlier experimental work [Pestov D, et al. (2007) Science 316:265-268] yield single-shot identification of a small sample of Bacillus subtilis endospores (approximately 10(4) spores). The results convey the utility of the technique and its potential for "on-the-fly" detection of biohazards, such as Bacillus anthracis. The application of optimized coherent anti-Stokes Raman scattering scheme to problems requiring chemical specificity and short signal acquisition times is demonstrated.
A lab-on-chip for biothreat detection using single-molecule DNA mapping.
Meltzer, Robert H; Krogmeier, Jeffrey R; Kwok, Lisa W; Allen, Richard; Crane, Bryan; Griffis, Joshua W; Knaian, Linda; Kojanian, Nanor; Malkin, Gene; Nahas, Michelle K; Papkov, Vyacheslav; Shaikh, Saad; Vyavahare, Kedar; Zhong, Qun; Zhou, Yi; Larson, Jonathan W; Gilmanshin, Rudolf
2011-03-07
Rapid, specific, and sensitive detection of airborne bacteria, viruses, and toxins is critical for biodefense, yet the diverse nature of the threats poses a challenge for integrated surveillance, as each class of pathogens typically requires different detection strategies. Here, we present a laboratory-on-a-chip microfluidic device (LOC-DLA) that integrates two unique assays for the detection of airborne pathogens: direct linear analysis (DLA) with unsurpassed specificity for bacterial threats and Digital DNA for toxins and viruses. The LOC-DLA device also prepares samples for analysis, incorporating upstream functions for concentrating and fractionating DNA. Both DLA and Digital DNA assays are single molecule detection technologies, therefore the assay sensitivities depend on the throughput of individual molecules. The microfluidic device and its accompanying operation protocols have been heavily optimized to maximize throughput and minimize the loss of analyzable DNA. We present here the design and operation of the LOC-DLA device, demonstrate multiplex detection of rare bacterial targets in the presence of 100-fold excess complex bacterial mixture, and demonstrate detection of picogram quantities of botulinum toxoid.
Asteroid detection using a single multi-wavelength CCD scan
NASA Astrophysics Data System (ADS)
Melton, Jonathan
2016-09-01
Asteroid detection is a topic of great interest due to the possibility of diverting possibly dangerous asteroids or mining potentially lucrative ones. Currently, asteroid detection is generally performed by taking multiple images of the same patch of sky separated by 10-15 minutes, then subtracting the images to find movement. However, this is time consuming because of the need to revisit the same area multiple times per night. This paper describes an algorithm that can detect asteroids using a single CCD camera scan, thus cutting down on the time and cost of an asteroid survey. The algorithm is based on the fact that some telescopes scan the sky at multiple wavelengths with a small time separation between the wavelength components. As a result, an object moving with sufficient speed will appear in different places in different wavelength components of the same image. Using image processing techniques we detect the centroids of points of light in the first component and compare these positions to the centroids in the other components using a nearest neighbor algorithm. The algorithm was used on a test set of 49 images obtained from the Sloan telescope in New Mexico and found 100% of known asteroids with only 3 false positives. This algorithm has the advantage of decreasing the amount of time required to perform an asteroid scan, thus allowing more sky to be scanned in the same amount of time or freeing a telescope for other pursuits.
Rodda, Gordon H.; Dean-Bradley, Kathryn; Campbell, Earl W.; Fritts, Thomas H.; Lardner, Bjorn; Yackel Adams, Amy A.; Reed, Robert N.
2015-01-01
To obtain quantitative information about population dynamics from counts of animals, the per capita detectabilities of each species must remain constant over the course of monitoring. We characterized lizard detection constancy for four species over 17 yr from a single site in northern Guam, a relatively benign situation because detection was relatively easy and we were able to hold constant the site, habitat type, species, season, and sampling method. We monitored two species of diurnal terrestrial skinks (Carlia ailanpalai [Curious Skink], Emoia caeruleocauda [Pacific Bluetailed Skink]) using glueboards placed on the ground in the shade for 3 h on rainless mornings, yielding 10,286 skink captures. We additionally monitored two species of nocturnal arboreal geckos (Hemidactylus frenatus [Common House Gecko]; Lepidodactylus lugubris [Mourning Gecko]) on the basis of 15,212 sightings. We compared these count samples to a series of complete censuses we conducted from four or more total removal plots (everything removed to mineral soil) totaling 400 m2(about 1% of study site) in each of the years 1995, 1999, and 2012, providing time-stamped quantification of detectability for each species. Unfortunately, the actual population trajectories taken by the four species were masked by unexplained variation in detectability. This observation of debilitating latent variability in lizard detectability under nearly ideal conditions undercuts our trust in population estimation techniques that fail to quantify venue-specific detectability, rely on pooled detection probability estimates, or assume that modulation in predefined environmental covariates suffices for estimating detectability.
Zhang, Jingqing; Boghossian, Ardemis A; Barone, Paul W; Rwei, Alina; Kim, Jong-Ho; Lin, Dahua; Heller, Daniel A; Hilmer, Andrew J; Nair, Nitish; Reuel, Nigel F; Strano, Michael S
2011-01-26
We report the selective detection of single nitric oxide (NO) molecules using a specific DNA sequence of d(AT)(15) oligonucleotides, adsorbed to an array of near-infrared fluorescent semiconducting single-walled carbon nanotubes (AT(15)-SWNT). While SWNT suspended with eight other variant DNA sequences show fluorescence quenching or enhancement from analytes such as dopamine, NADH, L-ascorbic acid, and riboflavin, d(AT)(15) imparts SWNT with a distinct selectivity toward NO. In contrast, the electrostatically neutral polyvinyl alcohol enables no response to nitric oxide, but exhibits fluorescent enhancement to other molecules in the tested library. For AT(15)-SWNT, a stepwise fluorescence decrease is observed when the nanotubes are exposed to NO, reporting the dynamics of single-molecule NO adsorption via SWNT exciton quenching. We describe these quenching traces using a birth-and-death Markov model, and the maximum likelihood estimator of adsorption and desorption rates of NO is derived. Applying the method to simulated traces indicates that the resulting error in the estimated rate constants is less than 5% under our experimental conditions, allowing for calibration using a series of NO concentrations. As expected, the adsorption rate is found to be linearly proportional to NO concentration, and the intrinsic single-site NO adsorption rate constant is 0.001 s(-1) μM NO(-1). The ability to detect nitric oxide quantitatively at the single-molecule level may find applications in new cellular assays for the study of nitric oxide carcinogenesis and chemical signaling, as well as medical diagnostics for inflammation.
Deconvolution single shot multibox detector for supermarket commodity detection and classification
NASA Astrophysics Data System (ADS)
Li, Dejian; Li, Jian; Nie, Binling; Sun, Shouqian
2017-07-01
This paper proposes an image detection model to detect and classify supermarkets shelves' commodity. Based on the principle of the features directly affects the accuracy of the final classification, feature maps are performed to combine high level features with bottom level features. Then set some fixed anchors on those feature maps, finally the label and the position of commodity is generated by doing a box regression and classification. In this work, we proposed a model named Deconvolutiuon Single Shot MultiBox Detector, we evaluated the model using 300 images photographed from real supermarket shelves. Followed the same protocol in other recent methods, the results showed that our model outperformed other baseline methods.
NASA Astrophysics Data System (ADS)
Ngo, Hoan T.; Gandra, Naveen; Fales, Andrew M.; Taylor, Steve M.; Vo-Dinh, Tuan
2017-02-01
Nucleic acid-based molecular diagnostics at the point-of-care (POC) and in resource-limited settings is still a challenge. We present a sensitive yet simple DNA detection method with single nucleotide polymorphism (SNP) identification capability. The detection scheme involves sandwich hybridization of magnetic beads conjugated with capture probes, target sequences, and ultrabright surface-enhanced Raman Scattering (SERS) nanorattles conjugated with reporter probes. Upon hybridization, the sandwich probes are concentrated at the detection focus controlled by a magnetic system for SERS measurements. The ultrabright SERS nanorattles, consisting of a core and a shell with resonance Raman reporters loaded in the gap space between the core and the shell, serve as SERS tags for ultrasensitive signal detection. Specific DNA sequences of the malaria parasite Plasmodium falciparum and dengue virus 1 (DENV1) were used as the model marker system. Detection limit of approximately 100 attomoles was achieved. Single nucleotide polymorphism (SNP) discrimination of wild type malaria DNA and mutant malaria DNA, which confers resistance to artemisinin drugs, was also demonstrated. The results demonstrate the molecular diagnostic potential of the nanorattle-based method to both detect and genotype infectious pathogens. The method's simplicity makes it a suitable candidate for molecular diagnosis at the POC and in resource-limited settings.
Real-time stop sign detection and distance estimation using a single camera
NASA Astrophysics Data System (ADS)
Wang, Wenpeng; Su, Yuxuan; Cheng, Ming
2018-04-01
In modern world, the drastic development of driver assistance system has made driving a lot easier than before. In order to increase the safety onboard, a method was proposed to detect STOP sign and estimate distance using a single camera. In STOP sign detection, LBP-cascade classifier was applied to identify the sign in the image, and the principle of pinhole imaging was based for distance estimation. Road test was conducted using a detection system built with a CMOS camera and software developed by Python language with OpenCV library. Results shows that that the proposed system reach a detection accuracy of maximum of 97.6% at 10m, a minimum of 95.00% at 20m, and 5% max error in distance estimation. The results indicate that the system is effective and has the potential to be used in both autonomous driving and advanced driver assistance driving systems.
Xiao, Zhuo; Lie, Puchang; Fang, Zhiyuan; Yu, Luxin; Chen, Junhua; Liu, Jie; Ge, Chenchen; Zhou, Xuemeng; Zeng, Lingwen
2012-09-04
A lateral flow biosensor for detection of single nucleotide polymorphism based on circular strand displacement reaction (CSDPR) has been developed. Taking advantage of high fidelity of T4 DNA ligase, signal amplification by CSDPR, and the optical properties of gold nanoparticles, this assay has reached a detection limit of 0.01 fM.
NASA Astrophysics Data System (ADS)
Radtke, J.; Sponner, J.; Jakobi, C.; Schneider, J.; Sommer, M.; Teichmann, T.; Ullrich, W.; Henniger, J.; Kormoll, T.
2018-01-01
Single photon detection applied to optically stimulated luminescence (OSL) dosimetry is a promising approach due to the low level of luminescence light and the known statistical behavior of single photon events. Time resolved detection allows to apply a variety of different and independent data analysis methods. Furthermore, using amplitude modulated stimulation impresses time- and frequency information into the OSL light and therefore allows for additional means of analysis. Considering the impressed frequency information, data analysis by using Fourier transform algorithms or other digital filters can be used for separating the OSL signal from unwanted light or events generated by other phenomena. This potentially lowers the detection limits of low dose measurements and might improve the reproducibility and stability of obtained data. In this work, an OSL system based on a single photon detector, a fast and accurate stimulation unit and an FPGA is presented. Different analysis algorithms which are applied to the single photon data are discussed.
Association between human papilloma virus/Epstein-Barr virus coinfection and oral carcinogenesis.
Jiang, Ru; Ekshyyan, Oleksandr; Moore-Medlin, Tara; Rong, Xiaohua; Nathan, Sean; Gu, Xin; Abreo, Fleurette; Rosenthal, Eben L; Shi, Mingxia; Guidry, Joseph T; Scott, Rona S; Hutt-Fletcher, Lindsey M; Nathan, Cherie-Ann O
2015-01-01
The recent epidemic of head and neck squamous cell carcinomas associated with human papilloma virus (HPV) has not addressed its association with lymphoid tissue in the oropharynx or the potential role of Epstein-Barr virus (EBV)/HPV coinfection. The prevalence of HPV and EBV infection/coinfection and CD21 mRNA expression were determined in normal and cancerous tissues from the oropharynx using in situ hybridization (ISH), p16, and quantitative reverse transcriptase PCR (qRT-PCR). The effects of coinfection on tumorigenicity were evaluated using proliferation and invasion assays. Normal oropharynx, tonsil, non-cancer base of tongue (BOT), and BOT from sleep apnea patients demonstrated EBV positivity ranging from 7% to 36% depending on the site and methods of detection used (qRT-PCR or ISH). Among non-malignant BOT samples, HPV positivity was noted only in 20%. The percent of tonsil and BOT cancers positive for HPV (up to 63% and 80%, respectively) or coinfected with HPV/EBV (up to 25% and 70%, respectively) were both significantly associated with cancer status. Notably, HPV/EBV coinfection was observed only in malignant tissue originating in lymphoid-rich oropharynx sites (tonsil, BOT). CD21 mRNA (the major EBV attachment receptor) was detected in tonsil and BOT epithelium, but not in soft-palate epithelium. Coinfected cell lines showed a significant increase in invasiveness (P < 0.01). There is a high prevalence of HPV/EBV infection and coinfection in BOT and tonsil cancers, possibly reflecting their origins in lymphoid-rich tissue. In vitro, cells modeling coinfection have an increased invasive potential. © 2014 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.
Tu, Yiheng; Huang, Gan; Hung, Yeung Sam; Hu, Li; Hu, Yong; Zhang, Zhiguo
2013-01-01
Event-related potentials (ERPs) are widely used in brain-computer interface (BCI) systems as input signals conveying a subject's intention. A fast and reliable single-trial ERP detection method can be used to develop a BCI system with both high speed and high accuracy. However, most of single-trial ERP detection methods are developed for offline EEG analysis and thus have a high computational complexity and need manual operations. Therefore, they are not applicable to practical BCI systems, which require a low-complexity and automatic ERP detection method. This work presents a joint spatial-time-frequency filter that combines common spatial patterns (CSP) and wavelet filtering (WF) for improving the signal-to-noise (SNR) of visual evoked potentials (VEP), which can lead to a single-trial ERP-based BCI.
Detecting Antigen-Specific T Cell Responses: From Bulk Populations to Single Cells.
Phetsouphanh, Chansavath; Zaunders, John James; Kelleher, Anthony Dominic
2015-08-12
A new generation of sensitive T cell-based assays facilitates the direct quantitation and characterization of antigen-specific T cell responses. Single-cell analyses have focused on measuring the quality and breadth of a response. Accumulating data from these studies demonstrate that there is considerable, previously-unrecognized, heterogeneity. Standard assays, such as the ICS, are often insufficient for characterization of rare subsets of cells. Enhanced flow cytometry with imaging capabilities enables the determination of cell morphology, as well as the spatial localization of the protein molecules within a single cell. Advances in both microfluidics and digital PCR have improved the efficiency of single-cell sorting and allowed multiplexed gene detection at the single-cell level. Delving further into the transcriptome of single-cells using RNA-seq is likely to reveal the fine-specificity of cellular events such as alternative splicing (i.e., splice variants) and allele-specific expression, and will also define the roles of new genes. Finally, detailed analysis of clonally related antigen-specific T cells using single-cell TCR RNA-seq will provide information on pathways of differentiation of memory T cells. With these state of the art technologies the transcriptomics and genomics of Ag-specific T cells can be more definitively elucidated.
Detecting Antigen-Specific T Cell Responses: From Bulk Populations to Single Cells
Phetsouphanh, Chansavath; Zaunders, John James; Kelleher, Anthony Dominic
2015-01-01
A new generation of sensitive T cell-based assays facilitates the direct quantitation and characterization of antigen-specific T cell responses. Single-cell analyses have focused on measuring the quality and breadth of a response. Accumulating data from these studies demonstrate that there is considerable, previously-unrecognized, heterogeneity. Standard assays, such as the ICS, are often insufficient for characterization of rare subsets of cells. Enhanced flow cytometry with imaging capabilities enables the determination of cell morphology, as well as the spatial localization of the protein molecules within a single cell. Advances in both microfluidics and digital PCR have improved the efficiency of single-cell sorting and allowed multiplexed gene detection at the single-cell level. Delving further into the transcriptome of single-cells using RNA-seq is likely to reveal the fine-specificity of cellular events such as alternative splicing (i.e., splice variants) and allele-specific expression, and will also define the roles of new genes. Finally, detailed analysis of clonally related antigen-specific T cells using single-cell TCR RNA-seq will provide information on pathways of differentiation of memory T cells. With these state of the art technologies the transcriptomics and genomics of Ag-specific T cells can be more definitively elucidated. PMID:26274954
Detecting Single-Nucleotides by Tunneling Current Measurements at Sub-MHz Temporal Resolution.
Morikawa, Takanori; Yokota, Kazumichi; Tanimoto, Sachie; Tsutsui, Makusu; Taniguchi, Masateru
2017-04-18
Label-free detection of single-nucleotides was performed by fast tunneling current measurements in a polar solvent at 1 MHz sampling rate using SiO₂-protected Au nanoprobes. Short current spikes were observed, suggestive of trapping/detrapping of individual nucleotides between the nanoelectrodes. The fall and rise features of the electrical signatures indicated signal retardation by capacitance effects with a time constant of about 10 microseconds. The high temporal resolution revealed current fluctuations, reflecting the molecular conformation degrees of freedom in the electrode gap. The method presented in this work may enable direct characterizations of dynamic changes in single-molecule conformations in an electrode gap in liquid.
Dittfeld, Anna; Gwizdek, Katarzyna; Koszowska, Aneta; Nowak, Justyna; Brończyk-Puzoń, Anna; Jagielski, Paweł; Oświęcimska, Joanna; Ziora, Katarzyna
2016-01-01
Orthorexia nervosa is an obsessive-compulsive focus on a "healthy" diet, which manifests as extreme concern about food content. This research was carried out in order to identify highly sensitive behaviours associated with eating a healthy diet. Assessment of risk of orthorexia nervosa among dietetics students compared to physiotherapy students. Presented research was conducted among 229 dietetic students and 201 physiotherapy students. It was based on general personal characteristics, the Bratman Test for Orthorexia (BOT) and questions posed by the authors. 26.6% of the dietetics students and only 14.9% of physiotherapy students were recognised as health food fanatics. The students of both dietetics (88.2%) and physiotherapy (52%) declared that they noticed changes in their attitude towards food after they had become students. It has been observed that these highly sensitive behaviours towards healthy eating are very common in both surveyed groups, however with a stronger tendency among the students of dietetics. © Polish Society for Pediatric Endocrinology and Diabetology.
Hiatt, Joseph B; Pritchard, Colin C; Salipante, Stephen J; O'Roak, Brian J; Shendure, Jay
2013-05-01
The detection and quantification of genetic heterogeneity in populations of cells is fundamentally important to diverse fields, ranging from microbial evolution to human cancer genetics. However, despite the cost and throughput advances associated with massively parallel sequencing, it remains challenging to reliably detect mutations that are present at a low relative abundance in a given DNA sample. Here we describe smMIP, an assay that combines single molecule tagging with multiplex targeted capture to enable practical and highly sensitive detection of low-frequency or subclonal variation. To demonstrate the potential of the method, we simultaneously resequenced 33 clinically informative cancer genes in eight cell line and 45 clinical cancer samples. Single molecule tagging facilitated extremely accurate consensus calling, with an estimated per-base error rate of 8.4 × 10(-6) in cell lines and 2.6 × 10(-5) in clinical specimens. False-positive mutations in the single molecule consensus base-calls exhibited patterns predominantly consistent with DNA damage, including 8-oxo-guanine and spontaneous deamination of cytosine. Based on mixing experiments with cell line samples, sensitivity for mutations above 1% frequency was 83% with no false positives. At clinically informative sites, we identified seven low-frequency point mutations (0.2%-4.7%), including BRAF p.V600E (melanoma, 0.2% alternate allele frequency), KRAS p.G12V (lung, 0.6%), JAK2 p.V617F (melanoma, colon, two lung, 0.3%-1.4%), and NRAS p.Q61R (colon, 4.7%). We anticipate that smMIP will be broadly adoptable as a practical and effective method for accurately detecting low-frequency mutations in both research and clinical settings.
Single photon detection using Geiger mode CMOS avalanche photodiodes
NASA Astrophysics Data System (ADS)
Lawrence, William G.; Stapels, Christopher; Augustine, Frank L.; Christian, James F.
2005-10-01
Geiger mode Avalanche Photodiodes fabricated using complementary metal-oxide-semiconductor (CMOS) fabrication technology combine high sensitivity detectors with pixel-level auxiliary circuitry. Radiation Monitoring Devices has successfully implemented CMOS manufacturing techniques to develop prototype detectors with active diameters ranging from 5 to 60 microns and measured detection efficiencies of up to 60%. CMOS active quenching circuits are included in the pixel layout. The actively quenched pixels have a quenching time less than 30 ns and a maximum count rate greater than 10 MHz. The actively quenched Geiger mode avalanche photodiode (GPD) has linear response at room temperature over six orders of magnitude. When operating in Geiger mode, these GPDs act as single photon-counting detectors that produce a digital output pulse for each photon with no associated read noise. Thermoelectrically cooled detectors have less than 1 Hz dark counts. The detection efficiency, dark count rate, and after-pulsing of two different pixel designs are measured and demonstrate the differences in the device operation. Additional applications for these devices include nuclear imaging and replacement of photomultiplier tubes in dosimeters.
Single-Use, Electricity-Free Amplification Device for Detection of HIV-1
Curtis, Kelly A.; Rudolph, Donna L.; Morrison, Daphne; Guelig, Dylan; Diesburg, Steven; McAdams, David; Burton, Robert A.; LaBarre, Paul; Owen, Michele
2016-01-01
Early and accurate diagnosis of HIV is key for the reduction of transmission and initiation of patient care. The availability of a rapid nucleic acid test (NAT) for use at the point-of-care (POC) will fill a gap in HIV diagnostics, improving the diagnosis of acute infection and HIV in infants born to infected mothers. In this study, we evaluated the performance of non-instrumented nucleic acid amplification, single-use disposable (NINA-SUD) devices for the detection of HIV-1 in whole blood using reverse-transcription, loop-mediated isothermal amplification (RT-LAMP) with lyophilized reagents. The NINA-SUD heating device harnesses the heat from an exothermic chemical reaction initiated by the addition of saline to magnesium iron powder. Reproducibility was demonstrated between NINA-SUD units and comparable, if not superior, performance for detecting clinical specimens was observed as compared to the thermal cycler. The stability of the lyophilized HIV-1 RT-LAMP reagents was also demonstrated following storage at −20, 4, 25, and 30°C for up to one month. The single-use, disposable NAT minimizes hands-on time and has the potential to facilitate HIV-1 testing in resource-limited settings or at the POC. PMID:27616198
GiniClust: detecting rare cell types from single-cell gene expression data with Gini index.
Jiang, Lan; Chen, Huidong; Pinello, Luca; Yuan, Guo-Cheng
2016-07-01
High-throughput single-cell technologies have great potential to discover new cell types; however, it remains challenging to detect rare cell types that are distinct from a large population. We present a novel computational method, called GiniClust, to overcome this challenge. Validation against a benchmark dataset indicates that GiniClust achieves high sensitivity and specificity. Application of GiniClust to public single-cell RNA-seq datasets uncovers previously unrecognized rare cell types, including Zscan4-expressing cells within mouse embryonic stem cells and hemoglobin-expressing cells in the mouse cortex and hippocampus. GiniClust also correctly detects a small number of normal cells that are mixed in a cancer cell population.
Cs2AgBiBr6 single-crystal X-ray detectors with a low detection limit
NASA Astrophysics Data System (ADS)
Pan, Weicheng; Wu, Haodi; Luo, Jiajun; Deng, Zhenzhou; Ge, Cong; Chen, Chao; Jiang, Xiaowei; Yin, Wan-Jian; Niu, Guangda; Zhu, Lujun; Yin, Lixiao; Zhou, Ying; Xie, Qingguo; Ke, Xiaoxing; Sui, Manling; Tang, Jiang
2017-11-01
Sensitive X-ray detection is crucial for medical diagnosis, industrial inspection and scientific research. The recently described hybrid lead halide perovskites have demonstrated low-cost fabrication and outstanding performance for direct X-ray detection, but they all contain toxic Pb in a soluble form. Here, we report sensitive X-ray detectors using solution-processed double perovskite Cs2AgBiBr6 single crystals. Through thermal annealing and surface treatment, we largely eliminate Ag+/Bi3+ disordering and improve the crystal resistivity, resulting in a detector with a minimum detectable dose rate as low as 59.7 nGyair s-1, comparable to the latest record of 0.036 μGyair s-1 using CH3NH3PbBr3 single crystals. Suppressed ion migration in Cs2AgBiBr6 permits relatively large external bias, guaranteeing efficient charge collection without a substantial increase in noise current and thus enabling the low detection limit.
Kreilinger, Alex; Hiebel, Hannah; Müller-Putz, Gernot R
2016-03-01
This work aimed to find and evaluate a new method for detecting errors in continuous brain-computer interface (BCI) applications. Instead of classifying errors on a single-trial basis, the new method was based on multiple events (MEs) analysis to increase the accuracy of error detection. In a BCI-driven car game, based on motor imagery (MI), discrete events were triggered whenever subjects collided with coins and/or barriers. Coins counted as correct events, whereas barriers were errors. This new method, termed ME method, combined and averaged the classification results of single events (SEs) and determined the correctness of MI trials, which consisted of event sequences instead of SEs. The benefit of this method was evaluated in an offline simulation. In an online experiment, the new method was used to detect erroneous MI trials. Such MI trials were discarded and could be repeated by the users. We found that, even with low SE error potential (ErrP) detection rates, feasible accuracies can be achieved when combining MEs to distinguish erroneous from correct MI trials. Online, all subjects reached higher scores with error detection than without, at the cost of longer times needed for completing the game. Findings suggest that ErrP detection may become a reliable tool for monitoring continuous states in BCI applications when combining MEs. This paper demonstrates a novel technique for detecting errors in online continuous BCI applications, which yields promising results even with low single-trial detection rates.
Morimoto, Atsushi; Mogami, Toshifumi; Watanabe, Masaru; Iijima, Kazuki; Akiyama, Yasuyuki; Katayama, Koji; Futami, Toru; Yamamoto, Nobuyuki; Sawada, Takeshi; Koizumi, Fumiaki; Koh, Yasuhiro
2015-01-01
Development of a reliable platform and workflow to detect and capture a small number of mutation-bearing circulating tumor cells (CTCs) from a blood sample is necessary for the development of noninvasive cancer diagnosis. In this preclinical study, we aimed to develop a capture system for molecular characterization of single CTCs based on high-density dielectrophoretic microwell array technology. Spike-in experiments using lung cancer cell lines were conducted. The microwell array was used to capture spiked cancer cells, and captured single cells were subjected to whole genome amplification followed by sequencing. A high detection rate (70.2%-90.0%) and excellent linear performance (R2 = 0.8189-0.9999) were noted between the observed and expected numbers of tumor cells. The detection rate was markedly higher than that obtained using the CellSearch system in a blinded manner, suggesting the superior sensitivity of our system in detecting EpCAM- tumor cells. Isolation of single captured tumor cells, followed by detection of EGFR mutations, was achieved using Sanger sequencing. Using a microwell array, we established an efficient and convenient platform for the capture and characterization of single CTCs. The results of a proof-of-principle preclinical study indicated that this platform has potential for the molecular characterization of captured CTCs from patients.
Glenn, Rachel; Dantus, Marcos
2016-01-07
Recent success with trace explosives detection based on the single ultrafast pulse excitation for remote stimulated Raman scattering (SUPER-SRS) prompts us to provide new results and a Perspective that describes the theoretical foundation of the strategy used for achieving the desired sensitivity and selectivity. SUPER-SRS provides fast and selective imaging while being blind to optical properties of the substrate such as color, texture, or laser speckle. We describe the strategy of combining coherent vibrational excitation with a reference pulse in order to detect stimulated Raman gain or loss. A theoretical model is used to reproduce experimental spectra and to determine the ideal pulse parameters for best sensitivity, selectivity, and resolution when detecting one or more compounds simultaneously.
NASA Astrophysics Data System (ADS)
Han, Jin-Hee; Kim, Hee-Joo; Lakshmana, Sudheendra; Gee, Shirley J.; Hammock, Bruce D.; Kennedy, Ian M.
2011-03-01
A nanoarray based-single molecule detection system was developed for detecting proteins with extremely high sensitivity. The nanoarray was able to effectively trap nanoparticles conjugated with biological sample into nanowells by integrating with an electrophoretic particle entrapment system (EPES). The nanoarray/EPES is superior to other biosensor using immunoassays in terms of saving the amounts of biological solution and enhancing kinetics of antibody binding due to reduced steric hindrance from the neighboring biological molecules. The nanoarray patterned onto a layer of PMMA and LOL on conductive and transparent indium tin oxide (ITO)-glass slide by using e-beam lithography. The suspension of 500 nm-fluorescent (green emission)-carboxylated polystyrene (PS) particles coated with protein-A followed by BDE 47 polyclonal antibody was added to the chip that was connected to the positive voltage. The droplet was covered by another ITO-coated-glass slide and connected to a ground terminal. After trapping the particles into the nanowells, the solution of different concentrations of anti-rabbit- IgG labeled with Alexa 532 was added for an immunoassay. A single molecule detection system could quantify the anti-rabbit IgG down to atto-mole level by counting photons emitted from the fluorescent dye bound to a single nanoparticle in a nanowell.
acdc – Automated Contamination Detection and Confidence estimation for single-cell genome data
Lux, Markus; Kruger, Jan; Rinke, Christian; ...
2016-12-20
A major obstacle in single-cell sequencing is sample contamination with foreign DNA. To guarantee clean genome assemblies and to prevent the introduction of contamination into public databases, considerable quality control efforts are put into post-sequencing analysis. Contamination screening generally relies on reference-based methods such as database alignment or marker gene search, which limits the set of detectable contaminants to organisms with closely related reference species. As genomic coverage in the tree of life is highly fragmented, there is an urgent need for a reference-free methodology for contaminant identification in sequence data. We present acdc, a tool specifically developed to aidmore » the quality control process of genomic sequence data. By combining supervised and unsupervised methods, it reliably detects both known and de novo contaminants. First, 16S rRNA gene prediction and the inclusion of ultrafast exact alignment techniques allow sequence classification using existing knowledge from databases. Second, reference-free inspection is enabled by the use of state-of-the-art machine learning techniques that include fast, non-linear dimensionality reduction of oligonucleotide signatures and subsequent clustering algorithms that automatically estimate the number of clusters. The latter also enables the removal of any contaminant, yielding a clean sample. Furthermore, given the data complexity and the ill-posedness of clustering, acdc employs bootstrapping techniques to provide statistically profound confidence values. Tested on a large number of samples from diverse sequencing projects, our software is able to quickly and accurately identify contamination. Results are displayed in an interactive user interface. Acdc can be run from the web as well as a dedicated command line application, which allows easy integration into large sequencing project analysis workflows. Acdc can reliably detect contamination in single-cell genome data. In
acdc – Automated Contamination Detection and Confidence estimation for single-cell genome data
DOE Office of Scientific and Technical Information (OSTI.GOV)
Lux, Markus; Kruger, Jan; Rinke, Christian
A major obstacle in single-cell sequencing is sample contamination with foreign DNA. To guarantee clean genome assemblies and to prevent the introduction of contamination into public databases, considerable quality control efforts are put into post-sequencing analysis. Contamination screening generally relies on reference-based methods such as database alignment or marker gene search, which limits the set of detectable contaminants to organisms with closely related reference species. As genomic coverage in the tree of life is highly fragmented, there is an urgent need for a reference-free methodology for contaminant identification in sequence data. We present acdc, a tool specifically developed to aidmore » the quality control process of genomic sequence data. By combining supervised and unsupervised methods, it reliably detects both known and de novo contaminants. First, 16S rRNA gene prediction and the inclusion of ultrafast exact alignment techniques allow sequence classification using existing knowledge from databases. Second, reference-free inspection is enabled by the use of state-of-the-art machine learning techniques that include fast, non-linear dimensionality reduction of oligonucleotide signatures and subsequent clustering algorithms that automatically estimate the number of clusters. The latter also enables the removal of any contaminant, yielding a clean sample. Furthermore, given the data complexity and the ill-posedness of clustering, acdc employs bootstrapping techniques to provide statistically profound confidence values. Tested on a large number of samples from diverse sequencing projects, our software is able to quickly and accurately identify contamination. Results are displayed in an interactive user interface. Acdc can be run from the web as well as a dedicated command line application, which allows easy integration into large sequencing project analysis workflows. Acdc can reliably detect contamination in single-cell genome data. In
Yi, Ping; Chen, Zhuqin; Zhao, Yan; Guo, Jianxin; Fu, Huabin; Zhou, Yuanguo; Yu, Lili; Li, Li
2009-03-01
The discovery of fetal DNA in maternal plasma has opened up an approach for noninvasive diagnosis. We have now assessed the possibility of detecting single-nucleotide differences between fetal and maternal DNA in maternal plasma by polymerase chain reaction (PCR)/ligase detection reaction((LDR)/capillary electrophoresis. PCR/LDR/capillary electrophoresis was applied to detect the genotype of c.454-397T>gene (ESR1) from experimental DNA models of maternal plasma at different sensitivity levels and 13 maternal plasma samples.alphaC in estrogen receptor. (1) Our results demonstrated that the technique could discriminate low abundance single-nucleotide mutation with a mutant/normal allele ratio up to 1:10 000. (2) Examination of ESR1 c.454-397T>C genotypes by using the method of restriction fragment length analysis was performed in 25 pregnant women, of whom 13 pregnant women had homozygous genotypes. The c.454-397T>C genotypes of paternally inherited fetal DNA in maternal plasma of these 13 women were detected by PCR/LDR/capillary electrophoresis, which were accordant with the results of umbilical cord blood. PCR/LDR/capillary electrophoresis has very high sensitivity to distinguish low abundance single nucleotide differences and can discriminate point mutations and single-nucleotide polymorphisms(SNPs) of paternally inherited fetal DNA in maternal plasma.
GAGG:ce single crystalline films: New perspective scintillators for electron detection in SEM.
Bok, Jan; Lalinský, Ondřej; Hanuš, Martin; Onderišinová, Zuzana; Kelar, Jakub; Kučera, Miroslav
2016-04-01
Single crystal scintillators are frequently used for electron detection in scanning electron microscopy (SEM). We report gadolinium aluminum gallium garnet (GAGG:Ce) single crystalline films as a new perspective scintillators for the SEM. For the first time, the epitaxial garnet films were used in a practical application: the GAGG:Ce scintillator was incorporated into a SEM scintillation electron detector and it showed improved image quality. In order to prove the GAGG:Ce quality accurately, the scintillation properties were examined using electron beam excitation and compared with frequently used scintillators in the SEM. The results demonstrate excellent emission efficiency of the GAGG:Ce single crystalline films together with their very fast scintillation decay useful for demanding SEM applications. Copyright © 2016 Elsevier B.V. All rights reserved.
NASA Astrophysics Data System (ADS)
Han, Minah; Jang, Hanjoo; Baek, Jongduk
2018-03-01
We investigate lesion detectability and its trends for different noise structures in single-slice and multislice CBCT images with anatomical background noise. Anatomical background noise is modeled using a power law spectrum of breast anatomy. Spherical signal with a 2 mm diameter is used for modeling a lesion. CT projection data are acquired by the forward projection and reconstructed by the Feldkamp-Davis-Kress algorithm. To generate different noise structures, two types of reconstruction filters (Hanning and Ram-Lak weighted ramp filters) are used in the reconstruction, and the transverse and longitudinal planes of reconstructed volume are used for detectability evaluation. To evaluate single-slice images, the central slice, which contains the maximum signal energy, is used. To evaluate multislice images, central nine slices are used. Detectability is evaluated using human and model observer studies. For model observer, channelized Hotelling observer (CHO) with dense difference-of-Gaussian (D-DOG) channels are used. For all noise structures, detectability by a human observer is higher for multislice images than single-slice images, and the degree of detectability increase in multislice images depends on the noise structure. Variation in detectability for different noise structures is reduced in multislice images, but detectability trends are not much different between single-slice and multislice images. The CHO with D-DOG channels predicts detectability by a human observer well for both single-slice and multislice images.
Single photon detection imaging of Cherenkov light emitted during radiation therapy
NASA Astrophysics Data System (ADS)
Adamson, Philip M.; Andreozzi, Jacqueline M.; LaRochelle, Ethan; Gladstone, David J.; Pogue, Brian W.
2018-03-01
Cherenkov imaging during radiation therapy has been developed as a tool for dosimetry, which could have applications in patient delivery verification or in regular quality audit. The cameras used are intensified imaging sensors, either ICCD or ICMOS cameras, which allow important features of imaging, including: (1) nanosecond time gating, (2) amplification by 103-104, which together allow for imaging which has (1) real time capture at 10-30 frames per second, (2) sensitivity at the level of single photon event level, and (3) ability to suppress background light from the ambient room. However, the capability to achieve single photon imaging has not been fully analyzed to date, and as such was the focus of this study. The ability to quantitatively characterize how a single photon event appears in amplified camera imaging from the Cherenkov images was analyzed with image processing. The signal seen at normal gain levels appears to be a blur of about 90 counts in the CCD detector, after going through the chain of photocathode detection, amplification through a microchannel plate PMT, excitation onto a phosphor screen and then imaged on the CCD. The analysis of single photon events requires careful interpretation of the fixed pattern noise, statistical quantum noise distributions, and the spatial spread of each pulse through the ICCD.
Feasibility of detecting Aflatoxin B1 in single maize kernels using hyperspectral imaging
USDA-ARS?s Scientific Manuscript database
The feasibility of detecting Aflatoxin B1 (AFB1) in single maize kernel inoculated with Aspergillus flavus conidia in the field, as well as its spatial distribution in the kernels, was assessed using near-infrared hyperspectral imaging (HSI) technique. Firstly, an image mask was applied to a pixel-b...
DOE Office of Scientific and Technical Information (OSTI.GOV)
Wetzel, D.; Shi, Y; Reffner, J
This reports the first detection of chemical heterogeneity in octenyl succinic anhydride modified single starch granules using a Fourier transform infrared (FT-IR) microspectroscopical technique that combines diffraction-limited infrared microspectroscopy with a step size that is less than the mask projected spot size focused on the plane of the sample. The high spatial resolution was achieved with the combination of the application of a synchrotron infrared source and the confocal image plane masking system of the double-pass single-mask Continuum{reg_sign} infrared microscope. Starch from grains such as corn and wheat exists in granules. The size of the granules depends on the plantmore » producing the starch. Granules used in this study typically had a median size of 15 {micro}m. In the production of modified starch, an acid anhydride typically is reacted with OH groups of the starch polymer. The resulting esterification adds the ester carbonyl (1723 cm{sup -1}) organic functional group to the polymer and the hydrocarbon chain of the ester contributes to the CH{sub 2} stretching vibration to enhance the intensity of the 2927 cm{sup -1} band. Detection of the relative modifying population on a single granule was accomplished by ratioing the baseline adjusted peak area of the carbonyl functional group to that of a carbohydrate band. By stepping a confocally defined infrared beam as small as 5 {micro}m x 5 {micro}m across a starch granule 1 {micro}m at a time in both the x and y directions, the heterogeneity is detected with the highest possible spatial resolution.« less
Zhou, Ruobo; Kunzelmann, Simone; Webb, Martin R.; Ha, Taekjip
2011-01-01
Single molecule detection is useful for characterizing nanoscale objects such as biological macromolecules, nano-particles and nano-devices with nano-meter spatial resolution. Fluorescence resonance energy transfer (FRET) is widely used as a single-molecule assay to monitor intramolecular dynamics in the distance range of 3–8 nm. Here we demonstrate that self-quenching of two rhodamine derivatives can be used to detect small conformational dynamics corresponding to sub-nanometer distance changes in a FRET-insensitive short range at the single molecule level. A ParM protein mutant labeled with two rhodamines works as a single molecule ADP sensor which has 20 times brighter fluorescence signal in the ADP bound state than the unbound state. Single molecule time trajectories show discrete transitions between fluorescence on and off states that can be directly ascribed to ADP binding and dissociation events. The conformational changes observed with 20:1 contrast are only 0.5 nm in magnitude and are between crystallographic distances of 1.6 nm and 2.1 nm, demonstrating exquisite sensitivity to short distance scale changes. The systems also allowed us to gain information on the photophysics of self-quenching induced by rhodamine stacking: (1) photobleaching of either of the two rhodamines eliminates quenching of the other rhodamine fluorophore and (2) photobleaching from the highly quenched, stacked state is only two-fold slower than from the unstacked state. PMID:22023515
Evidence of complex contagion of information in social media: An experiment using Twitter bots.
Mønsted, Bjarke; Sapieżyński, Piotr; Ferrara, Emilio; Lehmann, Sune
2017-01-01
It has recently become possible to study the dynamics of information diffusion in techno-social systems at scale, due to the emergence of online platforms, such as Twitter, with millions of users. One question that systematically recurs is whether information spreads according to simple or complex dynamics: does each exposure to a piece of information have an independent probability of a user adopting it (simple contagion), or does this probability depend instead on the number of sources of exposure, increasing above some threshold (complex contagion)? Most studies to date are observational and, therefore, unable to disentangle the effects of confounding factors such as social reinforcement, homophily, limited attention, or network community structure. Here we describe a novel controlled experiment that we performed on Twitter using 'social bots' deployed to carry out coordinated attempts at spreading information. We propose two Bayesian statistical models describing simple and complex contagion dynamics, and test the competing hypotheses. We provide experimental evidence that the complex contagion model describes the observed information diffusion behavior more accurately than simple contagion. Future applications of our results include more effective defenses against malicious propaganda campaigns on social media, improved marketing and advertisement strategies, and design of effective network intervention techniques.
Jain, Deepika; Sheth, Heena; Bender, Filitsa H; Weisbord, Steven D; Green, Jamie A
2014-01-01
Studies have shown that a single-item question might be useful in identifying patients with limited health literacy. However, the utility of the approach has not been studied in patients receiving maintenance peritoneal dialysis (PD). We assessed health literacy in a cohort of 31 PD patients by administering the Rapid Estimate of Adult Literacy in Medicine (REALM) and a single-item health literacy (SHL) screening question "How confident are you filling out medical forms by yourself?" (Extremely, Quite a bit, Somewhat, A little bit, or Not at all). To determine the accuracy of the single-item question for detecting limited health literacy, we performed sensitivity and specificity analyses of the SHL and plotted the area under the receiver operating characteristic (AUROC) curve using the REALM as a reference standard. Using a cut-off of "Somewhat" or less confident, the sensitivity of the SHL for detecting limited health literacy was 80%, and the specificity was 88%. The positive likelihood ratio was 6.9. The SHL had an AUROC of 0.79 (95% confidence interval: 0.52 to 1.00). Our results show that the SHL could be effective in detecting limited health literacy in PD patients.
Hiatt, Joseph B.; Pritchard, Colin C.; Salipante, Stephen J.; O'Roak, Brian J.; Shendure, Jay
2013-01-01
The detection and quantification of genetic heterogeneity in populations of cells is fundamentally important to diverse fields, ranging from microbial evolution to human cancer genetics. However, despite the cost and throughput advances associated with massively parallel sequencing, it remains challenging to reliably detect mutations that are present at a low relative abundance in a given DNA sample. Here we describe smMIP, an assay that combines single molecule tagging with multiplex targeted capture to enable practical and highly sensitive detection of low-frequency or subclonal variation. To demonstrate the potential of the method, we simultaneously resequenced 33 clinically informative cancer genes in eight cell line and 45 clinical cancer samples. Single molecule tagging facilitated extremely accurate consensus calling, with an estimated per-base error rate of 8.4 × 10−6 in cell lines and 2.6 × 10−5 in clinical specimens. False-positive mutations in the single molecule consensus base-calls exhibited patterns predominantly consistent with DNA damage, including 8-oxo-guanine and spontaneous deamination of cytosine. Based on mixing experiments with cell line samples, sensitivity for mutations above 1% frequency was 83% with no false positives. At clinically informative sites, we identified seven low-frequency point mutations (0.2%–4.7%), including BRAF p.V600E (melanoma, 0.2% alternate allele frequency), KRAS p.G12V (lung, 0.6%), JAK2 p.V617F (melanoma, colon, two lung, 0.3%–1.4%), and NRAS p.Q61R (colon, 4.7%). We anticipate that smMIP will be broadly adoptable as a practical and effective method for accurately detecting low-frequency mutations in both research and clinical settings. PMID:23382536
Atkinson, C; Emery, V C; Griffiths, P D
2014-02-01
Newborn screening for congenital cytomegalovirus (CCMV) using dried blood spots (DBS) has been proposed because many developed countries have DBS screening programmes in place for other diseases. The aim of this study was to develop a rapid, single tube nested polymerase chain reaction (PCR) method for enhanced detection of CMV from DBS compared to existing (single target) real time PCRs. The new method was compared with existing real time PCRs for sensitivity and specificity. Overall sensitivity of the single target PCR assays in both asymptomatic and symptomatic infants with laboratory confirmed congenital CMV was 69% (CMV PCR or culture positive before day 21 of life). In contrast, the single tube nested assay had an increased sensitivity of 81% with100% specificity. Overall the assay detected CMV from a DBS equivalent to an original blood sample which contained 500IU/ml. In conclusion this single tube nested methodology allows simultaneous amplification and detection of CMV DNA in 1.5h removing the associated contamination risk of a two step nested PCR. Owing to its increased sensitivity, it has the potential to be used as a screening assay and ultimately allow early identification and intervention for children with congenital CMV. Copyright © 2013 Elsevier B.V. All rights reserved.
Ultralow power trapping and fluorescence detection of single particles on an optofluidic chip.
Kühn, S; Phillips, B S; Lunt, E J; Hawkins, A R; Schmidt, H
2010-01-21
The development of on-chip methods to manipulate particles is receiving rapidly increasing attention. All-optical traps offer numerous advantages, but are plagued by large required power levels on the order of hundreds of milliwatts and the inability to act exclusively on individual particles. Here, we demonstrate a fully integrated electro-optical trap for single particles with optical excitation power levels that are five orders of magnitude lower than in conventional optical force traps. The trap is based on spatio-temporal light modulation that is implemented using networks of antiresonant reflecting optical waveguides. We demonstrate the combination of on-chip trapping and fluorescence detection of single microorganisms by studying the photobleaching dynamics of stained DNA in E. coli bacteria. The favorable size scaling facilitates the trapping of single nanoparticles on integrated optofluidic chips.
Colorimetric detection of UV light-induced single-strand DNA breaks using gold nanoparticles.
Kim, Joong Hyun; Chung, Chan Ho; Chung, Bong Hyun
2013-02-21
We developed a colorimetric method to specifically detect single-strand DNA breaks using gold nanoparticles. In our assay, broken DNA cannot stabilize gold nanoparticles to prevent salt-induced aggregation as good as intact DNA can, and this effect can be easily observed with the naked eye as a red-to-purple color change.
Detecting the Presence of Nora Virus in "Drosophila" Utilizing Single Fly RT-PCR
ERIC Educational Resources Information Center
Munn, Bethany; Ericson, Brad; Carlson, Darby J.; Carlson, Kimberly A.
2015-01-01
A single fly RT-PCR protocol has recently been developed to detect the presence of the persistent, horizontally transmitted Nora virus in "Drosophila." Wild-caught flies from Ohio were tested for the presence of the virus, with nearly one-fifth testing positive. The investigation presented can serve as an ideal project for biology…
Shachaf, Catherine M; Elchuri, Sailaja V; Koh, Ai Leen; Zhu, Jing; Nguyen, Lienchi N; Mitchell, Dennis J; Zhang, Jingwu; Swartz, Kenneth B; Sun, Lei; Chan, Selena; Sinclair, Robert; Nolan, Garry P
2009-01-01
Detection of single cell epitopes has been a mainstay of immunophenotyping for over three decades, primarily using fluorescence techniques for quantitation. Fluorescence has broad overlapping spectra, limiting multiplexing abilities. To expand upon current detection systems, we developed a novel method for multi-color immuno-detection in single cells using "Composite Organic-Inorganic Nanoparticles" (COINs) Raman nanoparticles. COINs are Surface-Enhanced Raman Scattering (SERS) nanoparticles, with unique Raman spectra. To measure Raman spectra in single cells, we constructed an automated, compact, low noise and sensitive Raman microscopy device (Integrated Raman BioAnalyzer). Using this technology, we detected proteins expressed on the surface in single cells that distinguish T-cells among human blood cells. Finally, we measured intracellular phosphorylation of Stat1 (Y701) and Stat6 (Y641), with results comparable to flow cytometry. Thus, we have demonstrated the practicality of applying COIN nanoparticles for measuring intracellular phosphorylation, offering new possibilities to expand on the current fluorescent technology used for immunoassays in single cells.
NASA Astrophysics Data System (ADS)
Schoendube, Jonas; Yusof, Azmi; Kalkandjiev, Kiril; Zengerle, Roland; Koltay, Peter
2015-02-01
This work presents the microfabrication and experimental evaluation of a dispenser chip, designed for isolation and printing of single cells by combining impedance sensing and drop-on-demand dispensing. The dispenser chip features 50 × 55 µm (width × height) microchannels, a droplet generator and microelectrodes for impedance measurements. The chip is fabricated by sandwiching a dry film photopolymer (TMMF) between a silicon and a Pyrex wafer. TMMF has been used to define microfluidic channels, to serve as low temperature (75 °C) bonding adhesive and as etch mask during 300 µm deep HF etching of the Pyrex wafer. Due to the novel fabrication technology involving the dry film resist, it became possible to fabricate facing electrodes at the top and bottom of the channel and to apply electrical impedance sensing for particle detection with improved performance. The presented microchip is capable of dispensing liquid and detecting microparticles via impedance measurement. Single polystyrene particles of 10 µm size could be detected with a mean signal amplitude of 0.39 ± 0.13 V (n=439 ) at particle velocities of up to 9.6 mm s-1 inside the chip.
Single-molecule detection of dihydroazulene photo-thermal reaction using break junction technique
NASA Astrophysics Data System (ADS)
Huang, Cancan; Jevric, Martyn; Borges, Anders; Olsen, Stine T.; Hamill, Joseph M.; Zheng, Jue-Ting; Yang, Yang; Rudnev, Alexander; Baghernejad, Masoud; Broekmann, Peter; Petersen, Anne Ugleholdt; Wandlowski, Thomas; Mikkelsen, Kurt V.; Solomon, Gemma C.; Brøndsted Nielsen, Mogens; Hong, Wenjing
2017-05-01
Charge transport by tunnelling is one of the most ubiquitous elementary processes in nature. Small structural changes in a molecular junction can lead to significant difference in the single-molecule electronic properties, offering a tremendous opportunity to examine a reaction on the single-molecule scale by monitoring the conductance changes. Here, we explore the potential of the single-molecule break junction technique in the detection of photo-thermal reaction processes of a photochromic dihydroazulene/vinylheptafulvene system. Statistical analysis of the break junction experiments provides a quantitative approach for probing the reaction kinetics and reversibility, including the occurrence of isomerization during the reaction. The product ratios observed when switching the system in the junction does not follow those observed in solution studies (both experiment and theory), suggesting that the junction environment was perturbing the process significantly. This study opens the possibility of using nano-structured environments like molecular junctions to tailor product ratios in chemical reactions.
DEsingle for detecting three types of differential expression in single-cell RNA-seq data.
Miao, Zhun; Deng, Ke; Wang, Xiaowo; Zhang, Xuegong
2018-04-24
The excessive amount of zeros in single-cell RNA-seq data include "real" zeros due to the on-off nature of gene transcription in single cells and "dropout" zeros due to technical reasons. Existing differential expression (DE) analysis methods cannot distinguish these two types of zeros. We developed an R package DEsingle which employed Zero-Inflated Negative Binomial model to estimate the proportion of real and dropout zeros and to define and detect 3 types of DE genes in single-cell RNA-seq data with higher accuracy. The R package DEsingle is freely available at https://github.com/miaozhun/DEsingle and is under Bioconductor's consideration now. zhangxg@tsinghua.edu.cn. Supplementary data are available at Bioinformatics online.
NASA Astrophysics Data System (ADS)
McCoy, Michael J.; Habermann, Timothy J.; Hanke, Craig J.; Adar, Fran; Campbell, William B.; Nithipatikom, Kasem
1999-04-01
We developed a confocal Raman microspectroscopic technique to study ligand-receptor bindings in single cells using Raman-labeled ligands and surface-enhanced Raman scattering (SERS). The adrenal zona glomerulosa (ZG) cells were used as a model in this study. ZG cells have a high density of angiotensin II (AII) receptors on the cellular membrane. There are two identified subtypes of AII receptors,namely AT1 and AT2 receptors. AII is a peptidic hormone, which upon binding to its receptors, stimulates the release of aldosterone from ZG cells. The cellular localization of these receptors subtypes was detected in single ZG cells by using immunocomplexation of receptors with specific antibodies and confocal Raman microspectroscopy. In the binding study, we used biotin-labeled AII to bind to its receptors in ZG cells. Then, avidin and Raman-labeled AII. The binding was measure directly on the single ZG cells. The results showed that the binding was displaced with unlabeled AII and specific AII antagonists. This is a rapid and sensitive technique for detection of cellular ligand bindings as well as antagonists screening in drug discovery.
Fast Detection of Airports on Remote Sensing Images with Single Shot MultiBox Detector
NASA Astrophysics Data System (ADS)
Xia, Fei; Li, HuiZhou
2018-01-01
This paper introduces a method for fast airport detection on remote sensing images (RSIs) using Single Shot MultiBox Detector (SSD). To our knowledge, this could be the first study which introduces an end-to-end detection model into airport detection on RSIs. Based on the common low-level features between natural images and RSIs, a convolution neural network trained on large amounts of natural images was transferred to tackle the airport detection problem with limited annotated data. To deal with the specific characteristics of RSIs, some related parameters in the SSD, such as the scales and layers, were modified for more accurate and rapider detection. The experiments show that the proposed method could achieve 83.5% Average Recall at 8 FPS on RSIs with the size of 1024*1024. In contrast to Faster R-CNN, an improvement on AP and speed could be obtained.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Kersell, Heath; Shirato, Nozomi; Cummings, Marvin
We use a nanofabricated scanning tunneling microscope tip as a detector to investigate local X-ray induced tunneling and electron emission from a single cobalt nanocluster on a Au(111) surface. The tip-detector is positioned a few angstroms above the nanocluster, and ramping the incident X-ray energy across the Co photoabsorption K-edge enables the detection of element specific electrons. Atomic-scale spatial dependent changes in the X-ray absorption cross section are directly measured by taking the X-ray induced current as a function of X-ray energy. From the measured sample and tip currents, element specific X-ray induced current components can be separated and therebymore » the corresponding yields for the X-ray induced processes of the single cobalt nanocluster can be determined. The detection of element specific synchrotron X-ray induced electrons of a single nanocluster opens a new avenue for materials characterization on a one particle at-a-time basis.« less
DOE Office of Scientific and Technical Information (OSTI.GOV)
Kersell, Heath; Shirato, Nozomi; Cummings, Marvin
Here, we use a nanofabricated scanning tunneling microscope tip as a detector to investigate local X-ray induced tunneling and electron emission from a single cobalt nanocluster on a Au(111) surface. The tip-detector is positioned a few angstroms above the nanocluster, and ramping the incident X-ray energy across the Co photoabsorption K-edge enables the detection of element specific electrons. Atomic-scale spatial dependent changes in the X-ray absorption cross section are directly measured by taking the X-ray induced current as a function of X-ray energy. From the measured sample and tip currents, element specific X-ray induced current components can be separated andmore » thereby the corresponding yields for the X-ray induced processes of the single cobalt nanocluster can be determined. The detection of element specific synchrotron X-ray induced electrons of a single nanocluster opens a new avenue for materials characterization on a one particle at-a-time basis.« less
Kersell, Heath; Shirato, Nozomi; Cummings, Marvin; ...
2017-09-05
Here, we use a nanofabricated scanning tunneling microscope tip as a detector to investigate local X-ray induced tunneling and electron emission from a single cobalt nanocluster on a Au(111) surface. The tip-detector is positioned a few angstroms above the nanocluster, and ramping the incident X-ray energy across the Co photoabsorption K-edge enables the detection of element specific electrons. Atomic-scale spatial dependent changes in the X-ray absorption cross section are directly measured by taking the X-ray induced current as a function of X-ray energy. From the measured sample and tip currents, element specific X-ray induced current components can be separated andmore » thereby the corresponding yields for the X-ray induced processes of the single cobalt nanocluster can be determined. The detection of element specific synchrotron X-ray induced electrons of a single nanocluster opens a new avenue for materials characterization on a one particle at-a-time basis.« less
SQUID-detected FMR: Resonance in single crystalline and polycrystalline yttrium iron garnet
NASA Astrophysics Data System (ADS)
O'Reilly, J. M.; Stamenov, P.
2018-04-01
Here two new techniques for the detection of broadband (100 MHz-20 GHz) ferromagnetic resonance (FMR)/ferrimagnetic resonance in single and poly-crystalline materials, which rely on SQUID-based gradiometry detection of small changes in the magnetisation, are developed. In the first method, small changes in the along-the-applied-field projection of the coupled magnetic moment (Δmz) are detected as the material is driven into resonance. Absolute measurement of the longitudinal component of the magnetisation and the resonance induced lowering of this moment makes estimation of the precession cone angle accessible, which is typically difficult to extract using conventional cavity or stripline based detection methods. The second method invokes the change in Δmz with the resonance-induced thermal heating (d/mz d T ). Magnetisation dynamics in bulk Y3Fe5O12 are observed over a broad range of experimental temperatures (4 K-400 K) and fields (10-500 mT). The inhomogeneous microwave excitation allows for the observation of higher magnetostatic modes and the convenient tracking of very broad resonances. The two SQUID-detection techniques when combined with conventional broadband vector network analyser-FMR, low-frequency magnetic susceptibility, and DC magnetometry, all easily realised, essentially concurrently, using the same module, greatly expand the amount of static and dynamic information accessible.
Rapid single cell detection of Staphylococcus aureus by aptamer-conjugated gold nanoparticles
Chang, Yi-Chung; Yang, Chia-Ying; Sun, Ruei-Lin; Cheng, Yi-Feng; Kao, Wei-Chen; Yang, Pan-Chyr
2013-01-01
Staphylococcus aureus is one of the most important human pathogens, causing more than 500,000 infections in the United States each year. Traditional methods for bacterial culture and identification take several days, wasting precious time for patients who are suffering severe bacterial infections. Numerous nucleic acid-based detection methods have been introduced to address this deficiency; however, the costs and requirement for expensive equipment may limit the widespread use of such technologies. Thus, there is an unmet demand of new platform technology to improve the bacterial detection and identification in clinical practice. In this study, we developed a rapid, ultra-sensitive, low cost, and non-polymerase chain reaction (PCR)-based method for bacterial identification. Using this method, which measures the resonance light-scattering signal of aptamer-conjugated gold nanoparticles, we successfully detected single S. aureus cell within 1.5 hours. This new platform technology may have potential to develop a rapid and sensitive bacterial testing at point-of-care. PMID:23689505
Rapid single cell detection of Staphylococcus aureus by aptamer-conjugated gold nanoparticles.
Chang, Yi-Chung; Yang, Chia-Ying; Sun, Ruei-Lin; Cheng, Yi-Feng; Kao, Wei-Chen; Yang, Pan-Chyr
2013-01-01
Staphylococcus aureus is one of the most important human pathogens, causing more than 500,000 infections in the United States each year. Traditional methods for bacterial culture and identification take several days, wasting precious time for patients who are suffering severe bacterial infections. Numerous nucleic acid-based detection methods have been introduced to address this deficiency; however, the costs and requirement for expensive equipment may limit the widespread use of such technologies. Thus, there is an unmet demand of new platform technology to improve the bacterial detection and identification in clinical practice. In this study, we developed a rapid, ultra-sensitive, low cost, and non-polymerase chain reaction (PCR)-based method for bacterial identification. Using this method, which measures the resonance light-scattering signal of aptamer-conjugated gold nanoparticles, we successfully detected single S. aureus cell within 1.5 hours. This new platform technology may have potential to develop a rapid and sensitive bacterial testing at point-of-care.
Bui, Phuc Tan; Nishino, Tomoaki; Shiigi, Hiroshi; Nagaoka, Tsutomu
2015-01-31
A DNA molecule was utilized as a probe tip to achieve single-molecule genetic diagnoses. Hybridization of the probe and target DNAs resulted in electron tunneling along the emergent double-stranded DNA. Simple stationary monitoring of the tunneling current leads to single-molecule DNA detection and discovery of base mismatches and methylation.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Howder, Collin R.; Bell, David M.; Anderson, Scott L.
2014-01-15
An instrument designed for non-destructive mass analysis of single trapped nanoparticles is described. The heart of the instrument is a 3D quadrupole (Paul) trap constructed to give optical access to the trap center along ten directions, allowing passage of lasers for particle heating and detection, particle injection, collection of scattered or fluorescent photons for particle detection and mass analysis, and collection of particles on TEM grids for analysis, as needed. Nanoparticles are injected using an electrospray ionization (ESI) source, and conditions are described for spraying and trapping polymer particles, bare metal particles, and ligand stabilized particles with masses ranging frommore » 200 kDa to >3 GDa. Conditions appropriate to ESI and injection of different types of particles are described. The instrument is equipped with two ion guides separating the ESI source and nanoparticle trap. The first ion guide is mostly to allow desolvation and differential pumping before the particles enter the trap section of the instrument. The second is a linear quadrupole guide, which can be operated in mass selective or mass band-pass modes to limit transmission to species with mass-to-charge ratios in the range of interest. With a little experience, the design allows injection of single particles into the trap upon demand.« less
Shachaf, Catherine M.; Elchuri, Sailaja V.; Koh, Ai Leen; Zhu, Jing; Nguyen, Lienchi N.; Mitchell, Dennis J.; Zhang, Jingwu; Swartz, Kenneth B.; Sun, Lei; Chan, Selena; Sinclair, Robert; Nolan, Garry P.
2009-01-01
Background Detection of single cell epitopes has been a mainstay of immunophenotyping for over three decades, primarily using fluorescence techniques for quantitation. Fluorescence has broad overlapping spectra, limiting multiplexing abilities. Methodology/Principal Findings To expand upon current detection systems, we developed a novel method for multi-color immuno-detection in single cells using “Composite Organic-Inorganic Nanoparticles” (COINs) Raman nanoparticles. COINs are Surface-Enhanced Raman Scattering (SERS) nanoparticles, with unique Raman spectra. To measure Raman spectra in single cells, we constructed an automated, compact, low noise and sensitive Raman microscopy device (Integrated Raman BioAnalyzer). Using this technology, we detected proteins expressed on the surface in single cells that distinguish T-cells among human blood cells. Finally, we measured intracellular phosphorylation of Stat1 (Y701) and Stat6 (Y641), with results comparable to flow cytometry. Conclusions/Significance Thus, we have demonstrated the practicality of applying COIN nanoparticles for measuring intracellular phosphorylation, offering new possibilities to expand on the current fluorescent technology used for immunoassays in single cells. PMID:19367337
Zhu, Jing; Ding, Yongshun; Liu, Xingti; Wang, Lei; Jiang, Wei
2014-09-15
Highly sensitive and selective detection strategy for single-base mutations is essential for risk assessment of malignancy and disease prognosis. In this work, a fluorescent detection method for single-base mutation was proposed based on high selectivity of toehold-mediated strand displacement reaction (TSDR) and powerful signal amplification capability of isothermal DNA amplification. A discrimination probe was specially designed with a stem-loop structure and an overhanging toehold domain. Hybridization between the toehold domain and the perfect matched target initiated the TSDR along with the unfolding of the discrimination probe. Subsequently, the target sequence acted as a primer to initiate the polymerization and nicking reactions, which released a great abundant of short sequences. Finally, the released strands were annealed with the reporter probe, launching another polymerization and nicking reaction to produce lots of G-quadruplex DNA, which could bind the N-methyl mesoporphyrin IX to yield an enhanced fluorescence response. However, when there was even a single base mismatch in the target DNA, the TSDR was suppressed and so subsequent isothermal DNA amplification and fluorescence response process could not occur. The proposed approach has been successfully implemented for the identification of the single-base mutant sequences in the human KRAS gene with a detection limit of 1.8 pM. Furthermore, a recovery of 90% was obtained when detecting the target sequence in spiked HeLa cells lysate, demonstrating the feasibility of this detection strategy for single-base mutations in biological samples. Copyright © 2014 Elsevier B.V. All rights reserved.
Low-cost coherent receiver for long-reach optical access network using single-ended detection.
Zhang, Xuebing; Li, Zhaohui; Li, Jianping; Yu, Changyuan; Lau, Alan Pak Tao; Lu, Chao
2014-09-15
A low-cost coherent receiver using two 2×3 optical hybrids and single-ended detection is proposed for long-reach optical access network. This structure can detect the two polarization components of polarization division multiplexing (PDM) signals. Polarization de-multiplexing and signal-to-signal beat interference (SSBI) cancellation are realized by using only three photodiodes. Simulation results for 40 Gb/s PDM-OFDM transmissions indicate that the low-cost coherent receiver has 3.2 dB optical signal-to-noise ratio difference compared with the theoretical value.
Using a nanopore for single molecule detection and single cell transfection.
Nelson, Edward M; Kurz, Volker; Shim, Jiwook; Timp, Winston; Timp, Gregory
2012-07-07
We assert that it is possible to trap and identify proteins, and even (conceivably) manipulate proteins secreted from a single cell (i.e. the secretome) through transfection via electroporation by exploiting the exquisite control over the electrostatic potential available in a nanopore. These capabilities may be leveraged for single cell analysis and transfection with single molecule resolution, ultimately enabling a careful scrutiny of tissue heterogeneity.
X-ray detection capability of a BaCl{sub 2} single crystal scintillator
DOE Office of Scientific and Technical Information (OSTI.GOV)
Koshimizu, Masanori; CREST, Japan Science and Technology Agency, Sanbancho, Chiyoda-ku, Tokyo 102-0075; Onodera, Kazuya
2012-01-15
The x-ray detection capability of a scintillation detector equipped with a BaCl{sub 2} single crystal was evaluated. The scintillation decay kinetics can be expressed by a sum of two exponential decay components. The fast and slow components have lifetimes of 1.5 and 85 ns, respectively. The total light output is 5% that of YAP:Ce. A subnanosecond timing resolution was obtained. The detection efficiency of a 67.41 keV x-ray is 87% for a detector equipped with a BaCl{sub 2} crystal 6-mm thick. Thus, excellent timing resolution and high detection efficiency can be simultaneously achieved. Additionally, luminescence decay characteristics under vacuum ultravioletmore » excitation have been investigated. Radiative decay of self-trapped excitons is thought to be responsible for the fast scintillation component.« less
Yang, Ming; Peng, Zhihui; Ning, Yi; Chen, Yongzhe; Zhou, Qin; Deng, Le
2013-05-22
In this paper, a panel of single-stranded DNA aptamers with high affinity and specificity against Salmonella Paratyphi A was selected from an enriched oligonucleotide pool by a whole-cell-Systematic Evolution of Ligands by Exponential Enrichment (SELEX) procedure, during which four other Salmonella serovars were used as counter-selection targets. It was determined through a fluorescence assay that the selected aptamers had high binding ability and specificity to this pathogen. The dissociation constant of these aptamers were up to nanomolar range, and aptamer Apt22 with the lowest Kd (47 ± 3 nM) was used in cell imaging experiments. To detect this bacteria with high specificity and cost-efficiently, a novel useful detection method was also constructed based on the noncovalent self-assembly of single-walled carbon nanotubes (SWNTs) and DNAzyme-labeled aptamer detection probes. The amounts of target bacteria could be quantified by exploiting chemoluminescence intensity changes at 420 nm and the detection limit of the method was 103 cfu/mL. This study demonstrated the applicability of Salmonella specific aptamers and their potential for use in the detection of Salmonella in food, clinical and environmental samples.
A Single-Chip CMOS Pulse Oximeter with On-Chip Lock-In Detection.
He, Diwei; Morgan, Stephen P; Trachanis, Dimitrios; van Hese, Jan; Drogoudis, Dimitris; Fummi, Franco; Stefanni, Francesco; Guarnieri, Valerio; Hayes-Gill, Barrie R
2015-07-14
Pulse oximetry is a noninvasive and continuous method for monitoring the blood oxygen saturation level. This paper presents the design and testing of a single-chip pulse oximeter fabricated in a 0.35 µm CMOS process. The chip includes photodiode, transimpedance amplifier, analogue band-pass filters, analogue-to-digital converters, digital signal processor and LED timing control. The experimentally measured AC and DC characteristics of individual circuits including the DC output voltage of the transimpedance amplifier, transimpedance gain of the transimpedance amplifier, and the central frequency and bandwidth of the analogue band-pass filters, show a good match (within 1%) with the circuit simulations. With modulated light source and integrated lock-in detection the sensor effectively suppresses the interference from ambient light and 1/f noise. In a breath hold and release experiment the single chip sensor demonstrates consistent and comparable performance to commercial pulse oximetry devices with a mean of 1.2% difference. The single-chip sensor enables a compact and robust design solution that offers a route towards wearable devices for health monitoring.
A Single-Chip CMOS Pulse Oximeter with On-Chip Lock-In Detection
He, Diwei; Morgan, Stephen P.; Trachanis, Dimitrios; van Hese, Jan; Drogoudis, Dimitris; Fummi, Franco; Stefanni, Francesco; Guarnieri, Valerio; Hayes-Gill, Barrie R.
2015-01-01
Pulse oximetry is a noninvasive and continuous method for monitoring the blood oxygen saturation level. This paper presents the design and testing of a single-chip pulse oximeter fabricated in a 0.35 µm CMOS process. The chip includes photodiode, transimpedance amplifier, analogue band-pass filters, analogue-to-digital converters, digital signal processor and LED timing control. The experimentally measured AC and DC characteristics of individual circuits including the DC output voltage of the transimpedance amplifier, transimpedance gain of the transimpedance amplifier, and the central frequency and bandwidth of the analogue band-pass filters, show a good match (within 1%) with the circuit simulations. With modulated light source and integrated lock-in detection the sensor effectively suppresses the interference from ambient light and 1/f noise. In a breath hold and release experiment the single chip sensor demonstrates consistent and comparable performance to commercial pulse oximetry devices with a mean of 1.2% difference. The single-chip sensor enables a compact and robust design solution that offers a route towards wearable devices for health monitoring. PMID:26184225
Detection of mandarin in orange juice by single-nucleotide polymorphism qPCR assay.
Aldeguer, Miriam; López-Andreo, María; Gabaldón, José A; Puyet, Antonio
2014-02-15
A dual-probe real time PCR (qPCR) DNA-based analysis was devised for the identification of mandarin in orange juice. A single nucleotide polymorphism at the trnL-trnF intergenic region of the chloroplast chromosome was confirmed in nine orange (Citrus sinensis) and thirteen commercial varieties of mandarin, including Citrus reticulata and Citrus unshiu species and a mandarin × tangelo hybrid. Two short minor-groove binding fluorescent probes targeting the polymorphic sequence were used in the dual-probe qPCR, which allowed the detection of both species in single-tube reactions. The similarity of PCR efficiencies allowed a simple estimation of the ratio mandarin/orange in the juice samples, which correlated to the measured difference of threshold cycle values for both probes. The limit of detection of the assay was 5% of mandarin in orange juice, both when the juice was freshly prepared (not from concentrate) or reconstituted from concentrate, which would allow the detection of fraudulently added mandarin juice. The possible use of the dual-probe system for quantitative measurements was also tested on fruit juice mixtures. qPCR data obtained from samples containing equal amounts of mandarin and orange juice revealed that the mandarin target copy number was approximately 2.6-fold higher than in orange juice. The use of a matrix-adapted control as calibrator to compensate the resulting C(T) bias allowed accurate quantitative measurements to be obtained. Copyright © 2013 Elsevier Ltd. All rights reserved.
Long-range depth profiling of camouflaged targets using single-photon detection
NASA Astrophysics Data System (ADS)
Tobin, Rachael; Halimi, Abderrahim; McCarthy, Aongus; Ren, Ximing; McEwan, Kenneth J.; McLaughlin, Stephen; Buller, Gerald S.
2018-03-01
We investigate the reconstruction of depth and intensity profiles from data acquired using a custom-designed time-of-flight scanning transceiver based on the time-correlated single-photon counting technique. The system had an operational wavelength of 1550 nm and used a Peltier-cooled InGaAs/InP single-photon avalanche diode detector. Measurements were made of human figures, in plain view and obscured by camouflage netting, from a stand-off distance of 230 m in daylight using only submilliwatt average optical powers. These measurements were analyzed using a pixelwise cross correlation approach and compared to analysis using a bespoke algorithm designed for the restoration of multilayered three-dimensional light detection and ranging images. This algorithm is based on the optimization of a convex cost function composed of a data fidelity term and regularization terms, and the results obtained show that it achieves significant improvements in image quality for multidepth scenarios and for reduced acquisition times.
Miki, Shigehito; Yamashita, Taro; Wang, Zhen; Terai, Hirotaka
2014-04-07
We present the characterization of two-dimensionally arranged 64-pixel NbTiN superconducting nanowire single-photon detector (SSPD) array for spatially resolved photon detection. NbTiN films deposited on thermally oxidized Si substrates enabled the high-yield production of high-quality SSPD pixels, and all 64 SSPD pixels showed uniform superconducting characteristics within the small range of 7.19-7.23 K of superconducting transition temperature and 15.8-17.8 μA of superconducting switching current. Furthermore, all of the pixels showed single-photon sensitivity, and 60 of the 64 pixels showed a pulse generation probability higher than 90% after photon absorption. As a result of light irradiation from the single-mode optical fiber at different distances between the fiber tip and the active area, the variations of system detection efficiency (SDE) in each pixel showed reasonable Gaussian distribution to represent the spatial distributions of photon flux intensity.
Rapid detection of single E. coli bacteria using a graphene-based field-effect transistor device.
Thakur, Bhawana; Zhou, Guihua; Chang, Jingbo; Pu, Haihui; Jin, Bing; Sui, Xiaoyu; Yuan, Xiaochen; Yang, Ching-Hong; Magruder, Matthew; Chen, Junhong
2018-07-01
Contamination of surface and drinking water due to the presence of Escherichia coli bacteria is a major cause of water-borne disease outbreak. To address unmet challenges for practical pathogen detection in contaminated samples, we report fabrication of thermally reduced graphene oxide-based field-effect transistor (rGO FET) passivated with an ultrathin layer of Al 2 O 3 for real-time detection of E. coli bacteria. The sensor could detect a single E. coli cell within 50 s in a 1 µL sample volume. The ultrathin layer of Al 2 O 3 acted as a barrier between rGO and potential interferents present in the sample. E. coli specific antibodies anchored on gold nanoparticles acted as probes for selective capture of E. coli. The high density of negative charge on the surface of E. coli cells strongly modulates the concentration of majority charge carriers in the rGO monolayer, thereby allowing real-time monitoring of E. coli concentration in a given sample. With a low detection limit of single cell, the FET sensor had a linear range of 1-100 CFU in 1 µL volume of sample (i.e., 10 3 to 10 5 CFU/ mL). The biosensor with good selectivity and rapid detection was further successfully demonstrated for E. coli sensing in river water. The rGO-based FET sensor provides a low cost and label-free approach, and can be mass produced for detection of a broad spectrum of pathogens in water or other liquid media. Copyright © 2018 Elsevier B.V. All rights reserved.
Grepstad, Jon Olav; Kaspar, Peter; Solgaard, Olav; Johansen, Ib-Rune; Sudbø, Aasmund S
2012-03-26
A sensor designed to detect bio-molecules is presented. The sensor exploits a planar 2D photonic crystal (PC) membrane with sub-micron thickness and through holes, to induce high optical fields that allow detection of nano-particles smaller than the diffraction limit of an optical microscope. We report on our design and fabrication of a PC membrane with a nano-particle trapped inside. We have also designed and built an imaging system where an optical microscope and a CCD camera are used to take images of the PC membrane. Results show how the trapped nano-particle appears as a bright spot in the image. In a first experimental realization of the imaging system, single particles with a radius of 75 nm can be detected.
Kollander, Barbro; Widemo, Fredrik; Ågren, Erik; Larsen, Erik H; Loeschner, Katrin
2017-03-01
This study investigated whether game meat may contain nanoparticles of lead from ammunition. Lead nanoparticles in the range 40 to 750 nm were detected by ICP-MS in single particle mode in game shot with lead-containing bullets. The median diameter of the detected nanoparticles was around 60 nm. The particle mass concentration ranged from 290 to 340 ng/g meat and the particle number concentrations from 27 to 50 million particles/g meat. The size limit of detection strongly depended on the level of dissolved lead and was in the range of 40 to 80 nm. In game meat sampled more than 10 cm away from the wound channel, no lead particles with a diameter larger than 40 nm were detected. In addition to dissolved lead in meat that originated from particulates, the presence of lead nano particles in game meat represents a hitherto unattended source of lead with a largely unknown toxicological impact to humans. Graphical Abstract Detection of lead nanoparticles in game meat by single particle ICP-MS following use of leadcontaining bullets.
Zhu, Zhi; Zhang, Wenhua; Leng, Xuefei; Zhang, Mingxia; Guan, Zhichao; Lu, Jiangquan; Yang, Chaoyong James
2012-10-21
Genetic alternations can serve as highly specific biomarkers to distinguish fatal bacteria or cancer cells from their normal counterparts. However, these mutations normally exist in very rare amount in the presence of a large excess of non-mutated analogs. Taking the notorious pathogen E. coli O157:H7 as the target analyte, we have developed an agarose droplet-based microfluidic ePCR method for highly sensitive, specific and quantitative detection of rare pathogens in the high background of normal bacteria. Massively parallel singleplex and multiplex PCR at the single-cell level in agarose droplets have been successfully established. Moreover, we challenged the system with rare pathogen detection and realized the sensitive and quantitative analysis of a single E. coli O157:H7 cell in the high background of 100,000 excess normal K12 cells. For the first time, we demonstrated rare pathogen detection through agarose droplet microfluidic ePCR. Such a multiplex single-cell agarose droplet amplification method enables ultra-high throughput and multi-parameter genetic analysis of large population of cells at the single-cell level to uncover the stochastic variations in biological systems.
Gaibani, Paolo; Campoli, Caterina; Lewis, Russell E; Volpe, Silvia Lidia; Scaltriti, Erika; Giannella, Maddalena; Pongolini, Stefano; Berlingeri, Andrea; Cristini, Francesco; Bartoletti, Michele; Tedeschi, Sara; Ambretti, Simone
2018-06-01
KPC-producing Klebsiella pneumoniae (KPC-Kp) represent a serious problem worldwide. Herein, we describe the evolution of ceftazidime/avibactam resistance by sequencing longitudinal clinical isolates from a patient with KPC-Kp bloodstream infection undergoing ceftazidime/avibactam treatment. WGS was performed on one ceftazidime/avibactam-susceptible KPC-Kp (BOT-CA-S) and two phenotypically different ceftazidime/avibactam-resistant KPC-Kp with low (BOT-CA-R) and high (BOT-EMO) carbapenem MICs. The population diversity was assessed by the frequency of allele mutations and population analysis profiles (PAPs). Phylogenetic analysis demonstrated clonal relatedness of the KPC-Kp isolates, all belonging to the clone ST1519. The D179Y mutation in blaKPC-3 was detected in both of the ceftazidime/avibactam-resistant KPC-Kp, whereas it was absent in the ceftazidime/avibactam-susceptible isolate. The mutation emerged independently in the two ceftazidime/avibactam-resistant isolates and was associated with a significant reduction in carbapenem MICs in BOT-CA-R, but not in BOT-EMO. WGS analysis revealed that the frequency of the D179Y mutation was 96.32% and 51.05% in BOT-CA-R and BOT-EMO, respectively. PAP results demonstrated that carbapenem resistance in BOT-EMO was due to the coexistence of mixed subpopulations harbouring WT and mutated blaKPC-3. A bacterial subpopulation with high ceftazidime/avibactam resistance for BOT-EMO KPC-Kp showed low carbapenem MICs, whereas a subpopulation with high meropenem resistance had a low MIC of ceftazidime/avibactam. Our analysis indicates that mixed subpopulations of ceftazidime/avibactam-resistant KPC-Kp emerge after ceftazidime/avibactam treatment. The evolution of different subpopulations that are highly resistant to ceftazidime/avibactam likely contributes to treatment failure, thereby highlighting the need for combination treatment strategies to limit selection of ceftazidime/avibactam-resistant KPC-Kp subpopulations.
Single-shot detection and direct control of carrier phase drift of midinfrared pulses.
Manzoni, Cristian; Först, Michael; Ehrke, Henri; Cavalleri, Andrea
2010-03-01
We introduce a scheme for single-shot detection and correction of the carrier-envelope phase (CEP) drift of femtosecond pulses at mid-IR wavelengths. Difference frequency mixing between the mid-IR field and a near-IR gate pulse generates a near-IR frequency-shifted pulse, which is then spectrally interfered with a replica of the gate pulse. The spectral interference pattern contains shot-to-shot information of the CEP of the mid-IR field, and it can be used for simultaneous correction of its slow drifts. We apply this technique to detect and compensate long-term phase drifts at 17 microm wavelength, reducing fluctuations to only 110 mrad over hours of operation.
Distributed multi-sensor particle filter for bearings-only tracking
NASA Astrophysics Data System (ADS)
Zhang, Jungen; Ji, Hongbing
2012-02-01
In this article, the classical bearings-only tracking (BOT) problem for a single target is addressed, which belongs to the general class of non-linear filtering problems. Due to the fact that the radial distance observability of the target is poor, the algorithm-based sequential Monte-Carlo (particle filtering, PF) methods generally show instability and filter divergence. A new stable distributed multi-sensor PF method is proposed for BOT. The sensors process their measurements at their sites using a hierarchical PF approach, which transforms the BOT problem from Cartesian coordinate to the logarithmic polar coordinate and separates the observable components from the unobservable components of the target. In the fusion centre, the target state can be estimated by utilising the multi-sensor optimal information fusion rule. Furthermore, the computation of a theoretical Cramer-Rao lower bound is given for the multi-sensor BOT problem. Simulation results illustrate that the proposed tracking method can provide better performances than the traditional PF method.
Lee, James W.; Thundat, Thomas G.
2006-04-25
An apparatus for carrying out the separation, detection, and/or counting of single molecules at nanometer scale. Molecular separation is achieved by driving single molecules through a microfluidic or nanofluidic medium using programmable and coordinated electric fields. In various embodiments, the fluidic medium is a strip of hydrophilic material on nonconductive hydrophobic surface, a trough produced by parallel strips of hydrophobic nonconductive material on a hydrophilic base, or a covered passageway produced by parallel strips of hydrophobic nonconductive material on a hydrophilic base together with a nonconductive cover on the parallel strips of hydrophobic nonconductive material. The molecules are detected and counted using nanoelectrode-gated electron tunneling methods, dielectric monitoring, and other methods.
Single-cell imaging techniques for the real-time detection of IP₃ in live cells.
Nelson, Carl P
2013-01-01
Inositol 1,4,5-trisphosphate (IP(3)) is a ubiquitous second messenger, derived from the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP(2)) by enzymes of the phospholipase C (PLC) family. Binding of IP(3) to its cognate receptor in the endoplasmic reticulum membrane leads to release of Ca(2+) into the cytoplasm, which is involved in the regulation of an array of cellular functions. Traditional techniques for the detection of IP(3) have required the extraction of a large number of cells, with limitations in the time resolution of changes in IP(3) and an inability to obtain detailed information on the dynamics of this second messenger in single cells. Recent progress in this field has led to the development of a number of genetically encoded fluorescent biosensors, which upon recombinant expression are able selectively to detect real-time changes in IP(3) in single live cells. In this chapter, I detail protocols for the expression, visualization (by confocol or fluorescence microscopy), and interpretation of data obtained with such biosensors expressed in mammalian cells.
Modulation and detection of single neuron activity using spin transfer nano-oscillators
NASA Astrophysics Data System (ADS)
Algarin, Jose Miguel; Ramaswamy, Bharath; Venuti, Lucy; Swierzbinski, Matthew; Villar, Pablo; Chen, Yu-Jin; Krivorotov, Ilya; Weinberg, Irving N.; Herberholz, Jens; Araneda, Ricardo; Shapiro, Benjamin; Waks, Edo
2017-09-01
The brain is a complex network of interconnected circuits that exchange electrical signals with each other. These electrical signals provide insight on how neural circuits code information, and give rise to sensations, thoughts, emotions and actions. Currents methods to detect and modulate these electrical signals use implanted electrodes or optical fields with light sensitive dyes in the brain. These techniques require complex surgeries or suffer low resolution. In this talk we explore a new method to both image and stimulate single neurons using spintronics. We propose using a Spin Transfer Nano-Oscillators (STNOs) as a nanoscale sensor that converts neuronal action potentials to microwave field oscillations that can be detected wirelessly by magnetic induction. We will describe our recent proof-of-concept demonstration of both detection and wireless modulation of neuronal activity using STNOs. For detection we use electrodes to connect a STNO to a lateral giant crayfish neuron. When we stimulate the neuron, the STNO responds to the neuronal activity with a corresponding microwave signal. For modulation, we stimulate the STNOs wirelessly using an inductively coupled solenoid. The STNO rectifies the induced microwave signal to produce a direct voltage. This direct voltage from the STNO, when applied in the vicinity of a mammalian neuron, changes the frequency of electrical signals produced by the neuron.
Torchetti, Enza Maria; Navarro, Beatriz; Di Serio, Francesco
2012-12-01
The spread of viroids belonging to the genus Pospiviroid (family Pospiviroidae), recorded recently in ornamentals and vegetables in several European countries, calls for fast, efficient and sensitive detection methods. Based on bioinformatics analyses of sequence identity among all pospiviroids, a digoxigenin-labeled polyprobe (POSPIprobe) was developed that, when tested by dot-blot and Northern-blot hybridization, detected Potato spindle tuber viroid, Citrus exocortis viroid, Columnea latent viroid, Mexican papita viroid, Tomato planta macho viroid, Tomato apical stunt viroid, Pepper chat fruit viroid and Chrysanthemum stunt viroid. The end-point detection limits of the POSPIprobe ranged from 5(-2) to 5(-4), and from 5(-1) to 5(-3) for nucleic acid preparations obtained by phenol extraction and silica-capture, respectively, similar to those of single probes. Based on sequence identity, the POSPIprobe is expected to detect also the two pospiviroid species not tested in this study (Tomato chlorotic dwarf viroid and Iresine viroid-1). Dot-blot assays with the POSPIprobe were validated by testing 68 samples from tomato, chrysanthemum and argyranthemum infected by different pospiviroids as revealed by RT-PCR, thus confirming the potential of this polyprobe for quarantine, certification and survey programs. Copyright © 2012 Elsevier B.V. All rights reserved.
Toward photostable multiplex analyte detection on a single mode planar optical waveguide
NASA Astrophysics Data System (ADS)
Mukundan, Harshini; Xie, Hongzhi; Anderson, Aaron; Grace, W. Kevin; Martinez, Jennifer S.; Swanson, Basil
2009-02-01
We have developed a waveguide-based optical biosensor for the sensitive and specific detection of biomarkers associated with disease. Our technology combines the superior optical properties of single-mode planar waveguides, the robust nature of functionalized self-assembled monolayer sensing films and the specificity of fluorescence sandwich immunoassays to detect biomarkers in complex biological samples such as serum, urine and sputum. We have previously reported the adaptation of our technology to the detection of biomarkers associated with breast cancer and anthrax. However, these approaches primarily used phospholipid bilayers as the functional film and organic dyes (ex: AlexaFluors) as the fluorescence reporter. Organic dyes are easily photodegraded and are not amenable to multiplexing because of their narrow Stokes' shift. Here we have developed strategies for conjugation of the detector antibodies with quantum dots for use in a multiplex detection platform. We have previously evaluated dihydroxylipoic acid quantum dots for the detection of a breast cancer biomarker. In this manuscript, we investigate the detection of the Bacillus anthracis protective antigen using antibodies conjugated with polymer-coated quantum dots. Kinetics of binding on the waveguide-based biosensor is reported. We compare the sensitivity of quantum dot labeled antibodies to those labeled with AlexaFluor and demonstrate the photostability of the former in our assay platform. In addition, we compare sulfydryl labeling of the antibody in the hinge region to that of nonspecific amine labeling. This is but the first step in developing a multiplex assay for such biomarkers on our waveguide platform.
Single Upconversion Nanoparticle-Bacterium Cotrapping for Single-Bacterium Labeling and Analysis.
Xin, Hongbao; Li, Yuchao; Xu, Dekang; Zhang, Yueli; Chen, Chia-Hung; Li, Baojun
2017-04-01
Detecting and analyzing pathogenic bacteria in an effective and reliable manner is crucial for the diagnosis of acute bacterial infection and initial antibiotic therapy. However, the precise labeling and analysis of bacteria at the single-bacterium level are a technical challenge but very important to reveal important details about the heterogeneity of cells and responds to environment. This study demonstrates an optical strategy for single-bacterium labeling and analysis by the cotrapping of single upconversion nanoparticles (UCNPs) and bacteria together. A single UCNP with an average size of ≈120 nm is first optically trapped. Both ends of a single bacterium are then trapped and labeled with single UCNPs emitting green light. The labeled bacterium can be flexibly moved to designated locations for further analysis. Signals from bacteria of different sizes are detected in real time for single-bacterium analysis. This cotrapping method provides a new approach for single-pathogenic-bacterium labeling, detection, and real-time analysis at the single-particle and single-bacterium level. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Su, Hui
2001-01-01
Laser-induced fluorescence detection is one of the most sensitive detection techniques and it has found enormous applications in various areas. The purpose of this research was to develop detection approaches based on laser-induced fluorescence detection in two different areas, heterogeneous catalysts screening and single cell study. First, the author introduced laser-induced imaging (LIFI) as a high-throughput screening technique for heterogeneous catalysts to explore the use of this high-throughput screening technique in discovery and study of various heterogeneous catalyst systems. This scheme is based on the fact that the creation or the destruction of chemical bonds alters the fluorescence properties ofmore » suitably designed molecules. By irradiating the region immediately above the catalytic surface with a laser, the fluorescence intensity of a selected product or reactant can be imaged by a charge-coupled device (CCD) camera to follow the catalytic activity as a function of time and space. By screening the catalytic activity of vanadium pentoxide catalysts in oxidation of naphthalene, they demonstrated LIFI has good detection performance and the spatial and temporal resolution needed for high-throughput screening of heterogeneous catalysts. The sample packing density can reach up to 250 x 250 subunits/cm 2 for 40-μm wells. This experimental set-up also can screen solid catalysts via near infrared thermography detection. In the second part of this dissertation, the author used laser-induced native fluorescence coupled with capillary electrophoresis (LINF-CE) and microscope imaging to study the single cell degranulation. On the basis of good temporal correlation with events observed through an optical microscope, they have identified individual peaks in the fluorescence electropherograms as serotonin released from the granular core on contact with the surrounding fluid.« less
Wen, Hong; Wang, Longye; Xie, Ping; Song, Liang; Tang, Jie; Liao, Runfa
2017-01-01
In this paper, we propose an adaptive single differential coherent detection (SDCD) scheme for the binary phase shift keying (BPSK) signals in IEEE 802.15.4 Wireless Sensor Networks (WSNs). In particular, the residual carrier frequency offset effect (CFOE) for differential detection is adaptively estimated, with only linear operation, according to the changing channel conditions. It was found that the carrier frequency offset (CFO) and chip signal-to-noise ratio (SNR) conditions do not need a priori knowledge. This partly benefits from that the combination of the trigonometric approximation sin−1(x)≈x and a useful assumption, namely, the asymptotic or high chip SNR, is considered for simplification of the full estimation scheme. Simulation results demonstrate that the proposed algorithm can achieve an accurate estimation and the detection performance can completely meet the requirement of the IEEE 802.15.4 standard, although with a little loss of reliability and robustness as compared with the conventional optimal single-symbol detector. PMID:29278404
Zhang, Gaoyuan; Wen, Hong; Wang, Longye; Xie, Ping; Song, Liang; Tang, Jie; Liao, Runfa
2017-12-26
In this paper, we propose an adaptive single differential coherent detection (SDCD) scheme for the binary phase shift keying (BPSK) signals in IEEE 802.15.4 Wireless Sensor Networks (WSNs). In particular, the residual carrier frequency offset effect (CFOE) for differential detection is adaptively estimated, with only linear operation, according to the changing channel conditions. It was found that the carrier frequency offset (CFO) and chip signal-to-noise ratio (SNR) conditions do not need a priori knowledge. This partly benefits from that the combination of the trigonometric approximation sin - 1 ( x ) ≈ x and a useful assumption, namely, the asymptotic or high chip SNR, is considered for simplification of the full estimation scheme. Simulation results demonstrate that the proposed algorithm can achieve an accurate estimation and the detection performance can completely meet the requirement of the IEEE 802.15.4 standard, although with a little loss of reliability and robustness as compared with the conventional optimal single-symbol detector.
Meemon, Panomsak; Rolland, Jannick P.
2010-01-01
Phase-Resolved Doppler Optical Coherence Tomography (PR-DOCT) allows visualization and characterization of the location, direction, velocity, and profile of flow activity embedded in a static sample structure. The detectable Velocity Dynamic Range (VDR) of each particular PR-DOCT system is governed by a detectable Doppler phase shift, a flow angle, and an acquisition time interval used to determine the Doppler phase shift. In general, the lower boundary of the detectable Doppler phase shift is limited by the phase stability of the system, while the upper boundary is limited by the π phase ambiguity. For a given range of detectable Doppler phase shift, shortening the acquisition duration will increase not only the maximum detectable velocity but unfortunately also the minimum detectable velocity, which may lead to the invisibility of a slow flow. In this paper, we present an alternative acquisition scheme for PR-DOCT that extends the lower limit of the velocity dynamic range, while maintaining the maximum detectable velocity, hence increasing the overall VDR of PR-DOCT system. The essence of the approach is to implement a technique of multi-scale measurement to simultaneously acquire multiple VDRs in a single measurement. We demonstrate an example of implementation of the technique in a dual VDR DOCT, where two Doppler maps having different detectable VDRs were simultaneously detected, processed, and displayed in real time. One was a fixed VDR DOCT capable of measuring axial velocity of up to 10.9 mm/s without phase unwrapping. The other was a variable VDR DOCT capable of adjusting its detectable VDR to reveal slow flow information down to 11.3 μm/s. The technique is shown to effectively extend the overall detectable VDR of the PR-DOCT system. Examples of real time Doppler imaging of an African frog tadpole are demonstrated using the dual-VDR DOCT system. PMID:21258521
Giri, Sidhartha; Rajan, Anand K; Kumar, Nirmal; Dhanapal, Pavithra; Venkatesan, Jayalakshmi; Iturriza-Gomara, Miren; Taniuchi, Mami; John, Jacob; Abraham, Asha Mary; Kang, Gagandeep
2017-08-01
Although, culture is considered the gold standard for poliovirus detection from stool samples, real-time PCR has emerged as a faster and more sensitive alternative. Detection of poliovirus from the stool of recently vaccinated children by culture, single and multiplex real-time PCR was compared. Of the 80 samples tested, 55 (68.75%) were positive by culture compared to 61 (76.25%) and 60 (75%) samples by the single and one step multiplex real-time PCR assays respectively. Real-time PCR (singleplex and multiplex) is more sensitive than culture for poliovirus detection in stool, although the difference was not statistically significant. © 2017 Wiley Periodicals, Inc.
Single vs. dual color fire detection systems: operational tradeoffs
NASA Astrophysics Data System (ADS)
Danino, Meir; Danan, Yossef; Sinvani, Moshe
2017-10-01
In attempt to supply a reasonable fire plume detection, multinational cooperation with significant capital is invested in the development of two major Infra-Red (IR) based fire detection alternatives, single-color IR (SCIR) and dual-color IR (DCIR). False alarm rate was expected to be high not only as a result of real heat sources but mainly due to the IR natural clutter especially solar reflections clutter. SCIR uses state-of-the-art technology and sophisticated algorithms to filter out threats from clutter. On the other hand, DCIR are aiming at using additional spectral band measurements (acting as a guard), to allow the implementation of a simpler and more robust approach for performing the same task. In this paper we present the basics of SCIR & DCIR architecture and the main differences between them. In addition, we will present the results from a thorough study conducted for the purpose of learning about the added value of the additional data available from the second spectral band. Here we consider the two CO2 bands of 4-5 micron and of 2.5-3 micron band as well as off peak band (guard). The findings of this study refer also to Missile warning systems (MWS) efficacy, in terms of operational value. We also present a new approach for tunable filter to such sensor.
Single-Cell Detection of Secreted Aβ and sAPPα from Human IPSC-Derived Neurons and Astrocytes.
Liao, Mei-Chen; Muratore, Christina R; Gierahn, Todd M; Sullivan, Sarah E; Srikanth, Priya; De Jager, Philip L; Love, J Christopher; Young-Pearse, Tracy L
2016-02-03
Secreted factors play a central role in normal and pathological processes in every tissue in the body. The brain is composed of a highly complex milieu of different cell types and few methods exist that can identify which individual cells in a complex mixture are secreting specific analytes. By identifying which cells are responsible, we can better understand neural physiology and pathophysiology, more readily identify the underlying pathways responsible for analyte production, and ultimately use this information to guide the development of novel therapeutic strategies that target the cell types of relevance. We present here a method for detecting analytes secreted from single human induced pluripotent stem cell (iPSC)-derived neural cells and have applied the method to measure amyloid β (Aβ) and soluble amyloid precursor protein-alpha (sAPPα), analytes central to Alzheimer's disease pathogenesis. Through these studies, we have uncovered the dynamic range of secretion profiles of these analytes from single iPSC-derived neuronal and glial cells and have molecularly characterized subpopulations of these cells through immunostaining and gene expression analyses. In examining Aβ and sAPPα secretion from single cells, we were able to identify previously unappreciated complexities in the biology of APP cleavage that could not otherwise have been found by studying averaged responses over pools of cells. This technique can be readily adapted to the detection of other analytes secreted by neural cells, which would have the potential to open new perspectives into human CNS development and dysfunction. We have established a technology that, for the first time, detects secreted analytes from single human neurons and astrocytes. We examine secretion of the Alzheimer's disease-relevant factors amyloid β (Aβ) and soluble amyloid precursor protein-alpha (sAPPα) and present novel findings that could not have been observed without a single-cell analytical platform. First, we
Kerman, Kagan; Saito, Masato; Tamiya, Eiichi
2008-08-01
Here we report an electrochemical biosensor that would allow for simple and rapid analysis of nucleic acids in combination with nuclease activity on nucleic acids and electroactive bionanoparticles. The detection of single-nucleotide polymorphisms (SNPs) using PNA probes takes advantage of the significant structural and physicochemical differences between the full hybrids and SNPs in PNA/DNA and DNA/DNA duplexes. Ferrocene-conjugated chitosan nanoparticles (Chi-Fc) were used as the electroactive indicator of hybridization. Chi-Fc had no affinity towards the neutral PNA probe immobilized on a gold electrode (AuE) surface. When the PNA probe on the electrode surface hybridized with a full-complementary target DNA, Chi-Fc electrostatically attached to the negatively-charged phosphate backbone of DNA on the surface and gave rise to a high electrochemical oxidation signal from ferrocene at approximately 0.30 V. Exposing the surface to a single-stranded DNA specific nuclease, Nuclease S1, was found to be very effective for removing the nonspecifically adsorbed SNP DNA. An SNP in the target DNA to PNA made it susceptible to the enzymatic digestion. After the enzymatic digestion and subsequent exposure to Chi-Fc, the presence of SNPs was determined by monitoring the changes in the electrical current response of Chi-Fc. The method provided a detection limit of 1 fM (S/N = 3) for the target DNA oligonucleotide. Additionally, asymmetric PCR was employed to detect the presence of genetically modified organism (GMO) in standard Roundup Ready soybean samples. PNA-mediated PCR amplification of real DNA samples was performed to detect SNPs related to alcohol dehydrogenase (ALDH). Chitosan nanoparticles are promising biomaterials for various analytical and pharmaceutical applications.
Detection of hail signatures from single-polarization C-band radar reflectivity
NASA Astrophysics Data System (ADS)
Kunz, Michael; Kugel, Petra I. S.
2015-02-01
Five different criteria that estimate hail signatures from single-polarization radar data are statistically evaluated over a 15-year period by categorical verification against loss data provided by a building insurance company. The criteria consider different levels or thresholds of radar reflectivity, some of them complemented by estimates of the 0 °C level or cloud top temperature. Applied to reflectivity data from a single C-band radar in southwest Germany, it is found that all criteria are able to reproduce most of the past damage-causing hail events. However, the criteria substantially overestimate hail occurrence by up to 80%, mainly due to the verification process using damage data. Best results in terms of highest Heidke Skill Score HSS or Critical Success Index CSI are obtained for the Hail Detection Algorithm (HDA) and the Probability of Severe Hail (POSH). Radar-derived hail probability shows a high spatial variability with a maximum on the lee side of the Black Forest mountains and a minimum in the broad Rhine valley.
NASA Astrophysics Data System (ADS)
Chen, Jiji; Wang, Chungang; Irudayaraj, Joseph
2009-07-01
A one-step rapid and ultrasensitive immunoassay capable of detecting proteins in blood serum is developed using gold nanoprobes and fluorescence correlation spectroscopy (FCS). In this approach we take advantage of the inherent photoluminescence property of gold nanoparticles (GNPs) to develop a fluorophore-free assay to observe binding entities by monitoring the diffusion of bound versus unbound molecules in a limited confocal volume. 40-nm GNPs conjugated separately with rabbit anti-IgG (Fc) and goat anti-IgG (Fab) when incubated in blood serum containing IgG forms a sandwich structure constituting dimers and oligomers that can be differentiated by to detect IgG in blood serum at a limit of detection (LOD) of 5 pg/ml. The novelty of integrating GNPs with FCS to develop a sensitive blood immunoassay brings single molecule methods one step closer to the clinic.
Gun muzzle flash detection using a CMOS single photon avalanche diode
NASA Astrophysics Data System (ADS)
Merhav, Tomer; Savuskan, Vitali; Nemirovsky, Yael
2013-10-01
Si based sensors, in particular CMOS Image sensors, have revolutionized low cost imaging systems but to date have hardly been considered as possible candidates for gun muzzle flash detection, due to performance limitations, and low SNR in the visible spectrum. In this study, a CMOS Single Photon Avalanche Diode (SPAD) module is used to record and sample muzzle flash events in the visible spectrum, from representative weapons, common on the modern battlefield. SPADs possess two crucial properties for muzzle flash imaging - Namely, very high photon detection sensitivity, coupled with a unique ability to convert the optical signal to a digital signal at the source pixel, thus practically eliminating readout noise. This enables high sampling frequencies in the kilohertz range without SNR degradation, in contrast to regular CMOS image sensors. To date, the SPAD has not been utilized for flash detection in an uncontrolled environment, such as gun muzzle flash detection. Gun propellant manufacturers use alkali salts to suppress secondary flashes ignited during the muzzle flash event. Common alkali salts are compounds based on Potassium or Sodium, with spectral emission lines around 769nm and 589nm, respectively. A narrow band filter around the Potassium emission doublet is used in this study to favor the muzzle flash signal over solar radiation. This research will demonstrate the SPAD's ability to accurately sample and reconstruct the temporal behavior of the muzzle flash in the visible wavelength under the specified imaging conditions. The reconstructed signal is clearly distinguishable from background clutter, through exploitation of flash temporal characteristics.
Nagarajan, Yagnesh; Rongala, Jay; Luang, Sukanya; Shadiac, Nadim; Sutton, Tim; Tyerman, Stephen D.; McPhee, Gordon; Voelcker, Nicolas H.; Lee, Jung-Goo
2016-01-01
Plant growth and survival depend upon the activity of membrane transporters that control the movement and distribution of solutes into, around, and out of plants. Although many plant transporters are known, their intrinsic properties make them difficult to study. In barley (Hordeum vulgare), the root anion-permeable transporter Bot1 plays a key role in tolerance to high soil boron, facilitating the efflux of borate from cells. However, its three-dimensional structure is unavailable and the molecular basis of its permeation function is unknown. Using an integrative platform of computational, biophysical, and biochemical tools as well as molecular biology, electrophysiology, and bioinformatics, we provide insight into the origin of transport function of Bot1. An atomistic model, supported by atomic force microscopy measurements, reveals that the protein folds into 13 transmembrane-spanning and five cytoplasmic α-helices. We predict a trimeric assembly of Bot1 and the presence of a Na+ ion binding site, located in the proximity of a pore that conducts anions. Patch-clamp electrophysiology of Bot1 detects Na+-dependent polyvalent anion transport in a Nernstian manner with channel-like characteristics. Using alanine scanning, molecular dynamics simulations, and transport measurements, we show that conductance by Bot1 is abolished by removal of the Na+ ion binding site. Our data enhance the understanding of the permeation functions of Bot1. PMID:26672067
Chen, Jiyun; Xu, Xiaomin; Huang, Zhimei; Luo, Yuan; Tang, Lijuan; Jiang, Jian-Hui
2018-01-02
A novel dNAD platform (BEAMing LAMP) by combining emulsion micro-reactors, single-molecule magnetic capture and on-bead loop-mediated isothermal amplification has been developed for DNA detection, which enables absolute and high-precision quantification of a target with a detection limit of 300 copies.
Automatic detection of respiration rate from ambulatory single-lead ECG.
Boyle, Justin; Bidargaddi, Niranjan; Sarela, Antti; Karunanithi, Mohan
2009-11-01
Ambulatory electrocardiography is increasingly being used in clinical practice to detect abnormal electrical behavior of the heart during ordinary daily activities. The utility of this monitoring can be improved by deriving respiration, which previously has been based on overnight apnea studies where patients are stationary, or the use of multilead ECG systems for stress testing. We compared six respiratory measures derived from a single-lead portable ECG monitor with simultaneously measured respiration air flow obtained from an ambulatory nasal cannula respiratory monitor. Ten controlled 1-h recordings were performed covering activities of daily living (lying, sitting, standing, walking, jogging, running, and stair climbing) and six overnight studies. The best method was an average of a 0.2-0.8 Hz bandpass filter and RR technique based on lengthening and shortening of the RR interval. Mean error rates with the reference gold standard were +/-4 breaths per minute (bpm) (all activities), +/-2 bpm (lying and sitting), and +/-1 breath per minute (overnight studies). Statistically similar results were obtained using heart rate information alone (RR technique) compared to the best technique derived from the full ECG waveform that simplifies data collection procedures. The study shows that respiration can be derived under dynamic activities from a single-lead ECG without significant differences from traditional methods.
Perez-Guaita, David; Andrew, Dean; Heraud, Philip; Beeson, James; Anderson, David; Richards, Jack; Wood, Bayden R
2016-06-23
New highly sensitive tools for malaria diagnostics are urgently needed to enable the detection of infection in asymptomatic carriers and patients with low parasitemia. In pursuit of a highly sensitive diagnostic tool that can identify parasite infections at the single cell level, we have been exploring Fourier transform infrared (FTIR) microscopy using a Focal Plane Array (FPA) imaging detector. Here we report for the first time the application of a new optic configuration developed by Agilent that incorporates 25× condenser and objective Cassegrain optics with a high numerical aperture (NA = 0.81) along with additional high magnification optics within the microscope to provide 0.66 micron pixel resolution (total IR system magnification of 61×) to diagnose malaria parasites at the single cell level on a conventional glass microscope slide. The high quality images clearly resolve the parasite's digestive vacuole demonstrating sub-cellular resolution using this approach. Moreover, we have developed an algorithm that first detects the cells in the infrared image, and secondly extracts the average spectrum. The average spectrum is then run through a model based on Partial Least Squares-Discriminant Analysis (PLS-DA), which diagnoses unequivocally the infected from normal cells. The high quality images, and the fact this measurement can be achieved without a synchrotron source on a conventional glass slide, shows promise as a potential gold standard for malaria detection at the single cell level.
Single-channel EEG-based mental fatigue detection based on deep belief network.
Pinyi Li; Wenhui Jiang; Fei Su
2016-08-01
Mental fatigue has a pernicious influence on road and work place safety as well as a negative symptom of many acute and chronic illnesses, since the ability of concentrating, responding and judging quickly decreases during the fatigue or drowsiness stage. Electroencephalography (EEG) has been proven to be a robust physiological indicator of human cognitive state over the last few decades. But most existing EEG-based fatigue detection methods have poor performance in accuracy. This paper proposed a single-channel EEG-based mental fatigue detection method based on Deep Belief Network (DBN). The fused nonliear features from specified sub-bands and dynamic analysis, a total of 21 features are extracted as the input of the DBN to discriminate three classes of mental state including alert, slight fatigue and severe fatigue. Experimental results show the good performance of the proposed model comparing with those state-of-art methods.
Gu, Li-Qun; Ding, Shu; Gao, Changlu
2011-01-01
The molecular-scale pore structure, called nanopore, can be formed from protein ion channels by genetic engineering or fabricated on solid substrates using fashion nanotechnology. Target molecules in interaction with the functionalized lumen of nanopore, can produce characteristic changes in the pore conductance, which act as fingerprints, allowing us to identify single molecules and simultaneously quantify each target species in the mixture. Nanopore sensors have been created for tremendous biomedical detections, with targets ranging from metal ions, drug compounds and cellular second messengers, to proteins and DNAs. Here we will review our recent discoveries with a lab-in-hand glass nanopore: single-molecule discrimination of chiral enantiomers with a trapped cyclodextrin, and sensing of bioterrorist agent ricin. PMID:19964179
Luo, Jun; Li, Junhua; Yang, Hang; Yu, Junping; Wei, Hongping
2017-10-01
Accurate and rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) is needed to screen MRSA carriers and improve treatment. The current widely used duplex PCR methods are not able to differentiate MRSA from coexisting methicillin-susceptible S. aureus (MSSA) or other methicillin-resistant staphylococci. In this study, we aimed to develop a direct method for accurate and rapid detection of MRSA in clinical samples from open environments, such as nasal swabs. The new molecular assay is based on detecting the cooccurrence of nuc and mecA markers in a single bacterial cell by utilizing droplet digital PCR (ddPCR) with the chimeric lysin ClyH for cell lysis. The method consists of (i) dispersion of an intact single bacterium into nanoliter droplets, (ii) temperature-controlled release of genomic DNA (gDNA) by ClyH at 37°C, and (iii) amplification and detection of the markers ( nuc and mecA ) using standard TaqMan chemistries with ddPCR. Results were analyzed based on MRSA index ratios used for indicating the presence of the duplex-positive markers in droplets. The method was able to achieve an absolute limit of detection (LOD) of 2,900 CFU/ml for MRSA in nasal swabs spiked with excess amounts of Escherichia coli , MSSA, and other mecA -positive bacteria within 4 h. Initial testing of 104 nasal swabs showed that the method had 100% agreement with the standard culture method, while the normal duplex qPCR method had only about 87.5% agreement. The single-bacterium duplex ddPCR assay is rapid and powerful for more accurate detection of MRSA directly from clinical specimens. Copyright © 2017 American Society for Microbiology.
Luo, Jun; Li, Junhua; Yang, Hang; Yu, Junping
2017-01-01
ABSTRACT Accurate and rapid identification of methicillin-resistant Staphylococcus aureus (MRSA) is needed to screen MRSA carriers and improve treatment. The current widely used duplex PCR methods are not able to differentiate MRSA from coexisting methicillin-susceptible S. aureus (MSSA) or other methicillin-resistant staphylococci. In this study, we aimed to develop a direct method for accurate and rapid detection of MRSA in clinical samples from open environments, such as nasal swabs. The new molecular assay is based on detecting the cooccurrence of nuc and mecA markers in a single bacterial cell by utilizing droplet digital PCR (ddPCR) with the chimeric lysin ClyH for cell lysis. The method consists of (i) dispersion of an intact single bacterium into nanoliter droplets, (ii) temperature-controlled release of genomic DNA (gDNA) by ClyH at 37°C, and (iii) amplification and detection of the markers (nuc and mecA) using standard TaqMan chemistries with ddPCR. Results were analyzed based on MRSA index ratios used for indicating the presence of the duplex-positive markers in droplets. The method was able to achieve an absolute limit of detection (LOD) of 2,900 CFU/ml for MRSA in nasal swabs spiked with excess amounts of Escherichia coli, MSSA, and other mecA-positive bacteria within 4 h. Initial testing of 104 nasal swabs showed that the method had 100% agreement with the standard culture method, while the normal duplex qPCR method had only about 87.5% agreement. The single-bacterium duplex ddPCR assay is rapid and powerful for more accurate detection of MRSA directly from clinical specimens. PMID:28724560
Sacchetti, P; Zvara, P; Plante, M K
1999-06-01
The information revolution triggered by the rapid growth of the Internet has allowed healthcare providers and patients to access a rapidly expanding volume of information. To address the quality of this information, a survey of the data on a single urology-related topic available on the Internet was performed. The search on the World Wide Web (Web) was performed using the search engine HotBot and search directory Yahoo. The Web pages were assessed according to their relevancy to the topic chosen. Relevance rates were derived from the number of relevant sites divided by the total number of sites found. Relevant sites were subsequently ranked for quality on the basis of their accuracy, comprehensiveness, and objectivity. HotBot was then subsequently divided by domain, with each assessed separately. Yahoo was analyzed in its entirety. The resources were then compared for relevance and quality of information. When using the keyword "Viagra," HotBot responded with 15,109 hits. Yahoo presented 51 hits under the category, "Health: Pharmacy: Drugs and Medications: Specific Drugs and Medications: Viagra (Sildenafil)." The relevance rate for the first 50 hits in the search engine HotBot was 0.08. The relevance rates for the edu and org domains found by HotBot were 0.22 and 0.24, respectively; those for com and net were both 0.10. The relevance rate for the search directory Yahoo was 0.20. For relevant sites, the quality of the information presented was significantly higher in the Yahoo and in the HotBot domains hosted by nonprofit organizations when compared with HotBot in general and with its commercially oriented domains. HotBot overall was found to contain seven excellent sites, of which only three were found within Yahoo. Although the medical information available on the Web has proliferated at a remarkable rate, the number of Web sites providing complete, nonbiased information continues to represent only a small portion of the total. We have shown that the search
Bloch oscillating transistor as the readout element for hot electron bolometers
NASA Astrophysics Data System (ADS)
Hassel, Juha; Seppä, Heikki; Lindell, Rene; Hakonen, Pertti
2004-10-01
In this paper we analyse the properties of the Bloch oscillating transistor as a preamplifier in cryogenic devices. We consider here especially the readout of hot electron bolometers (HEBs) based on Normal-Superconductor-Insulator tunnel junctions, but the results also apply more generally. We show that one can get an equivalent noise voltage below 1 nV/√Hz with a single BOT. By using N BOTs in a parallel array configuration, a further reduction by factor √N may be achieved.
NASA Astrophysics Data System (ADS)
Falzone, Nadia; Myhra, Sverre; Chakalova, Radka; Hill, Mark A.; Thomson, James; Vallis, Katherine A.
2013-11-01
The interactions between energetic ions and biological and/or organic target materials have recently attracted theoretical and experimental attention, due to their implications for detector and device technologies, and for therapeutic applications. Most of the attention has focused on detection of the primary ionization tracks, and their effects, while recoil target atom tracks remain largely unexplored. Detection of tracks by a negative tone photoresist (SU-8), followed by standard development, in combination with analysis by atomic force microscopy, shows that both primary and recoil tracks are revealed as conical spikes, and can be characterized at high spatial resolution. The methodology has the potential to provide detailed information about single impact events, which may lead to more effective and informative detector technologies and advanced therapeutic procedures. In comparison with current characterization methods the advantageous features include: greater spatial resolution by an order of magnitude (20 nm) detection of single primary and associated recoil tracks; increased range of fluence (to 2.5 × 109 cm-2) sensitivity to impacts at grazing angle incidence; and better definition of the lateral interaction volume in target materials.
Sensitive detection of multiple pathogens using a single DNA probe.
Nordin, Noordiana; Yusof, Nor Azah; Abdullah, Jaafar; Radu, Son; Hushiarian, Roozbeh
2016-12-15
A simple but promising electrochemical DNA nanosensor was designed, constructed and applied to differentiate a few food-borne pathogens. The DNA probe was initially designed to have a complementary region in Vibrio parahaemolyticus (VP) genome and to make different hybridization patterns with other selected pathogens. The sensor was based on a screen printed carbon electrode (SPCE) modified with polylactide-stabilized gold nanoparticles (PLA-AuNPs) and methylene blue (MB) was employed as the redox indicator binding better to single-stranded DNA. The immobilization and hybridization events were assessed using differential pulse voltammetry (DPV). The fabricated biosensor was able to specifically distinguish complementary, non-complementary and mismatched oligonucleotides. DNA was measured in the range of 2.0×10(-9)-2.0×10(-13)M with a detection limit of 5.3×10(-12)M. The relative standard deviation for 6 replications of DPV measurement of 0.2µM complementary DNA was 4.88%. The fabricated DNA biosensor was considered stable and portable as indicated by a recovery of more than 80% after a storage period of 6 months at 4-45°C. Cross-reactivity studies against various food-borne pathogens showed a reliably sensitive detection of VP. Copyright © 2016 Elsevier B.V. All rights reserved.
Collins, Valerie M; Daly, Donna M; Liaskos, Marina; McKay, Neil G; Sellers, Donna; Chapple, Christopher; Grundy, David
2013-11-01
To investigate the direct effect of onabotulinumtoxinA (OnaBotA) on bladder afferent nerve activity and release of ATP and acetylcholine (ACh) from the urothelium. Bladder afferent nerve activity was recorded using an in vitro mouse preparation enabling simultaneous recordings of afferent nerve firing and intravesical pressure during bladder distension. Intraluminal and extraluminal ATP, ACh, and nitric oxide (NO) release were measured using the luciferin-luciferase and Amplex(®) Red assays (Molecular Probes, Carlsbad, CA, USA), and fluorometric assay kit, respectively. OnaBotA (2U), was applied intraluminally, during bladder distension, and its effect was monitored for 2 h after application. Whole-nerve activity was analysed to classify the single afferent units responding to physiological (low-threshold [LT] afferent <15 mmHg) and supra-physiological (high-threshold [HT] afferent >15 mmHg) distension pressures. Bladder distension evoked reproducible pressure-dependent increases in afferent nerve firing. After exposure to OnaBotA, both LT and HT afferent units were significantly attenuated. OnaBotA also significantly inhibited ATP release from the urothelium and increased NO release. These data indicate that OnaBotA attenuates the bladder afferent nerves involved in micturition and bladder sensation, suggesting that OnaBotA may exert its clinical effects on urinary urgency and the other symptoms of overactive bladder syndrome through its marked effect on afferent nerves. © 2013 The Authors. BJU International © 2013 BJU International.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Martinenghi, E., E-mail: edoardo.martinenghi@polimi.it; Di Sieno, L.; Contini, D.
2016-07-15
We present the design and preliminary characterization of the first detection module based on Silicon Photomultiplier (SiPM) tailored for single-photon timing applications. The aim of this work is to demonstrate, thanks to the design of a suitable module, the possibility to easily exploit SiPM in many applications as an interesting detector featuring large active area, similarly to photomultipliers tubes, but keeping the advantages of solid state detectors (high quantum efficiency, low cost, compactness, robustness, low bias voltage, and insensitiveness to magnetic field). The module integrates a cooled SiPM with a total photosensitive area of 1 mm{sup 2} together with themore » suitable avalanche signal read-out circuit, the signal conditioning, the biasing electronics, and a Peltier cooler driver for thermal stabilization. It is able to extract the single-photon timing information with resolution better than 100 ps full-width at half maximum. We verified the effective stabilization in response to external thermal perturbations, thus proving the complete insensitivity of the module to environment temperature variations, which represents a fundamental parameter to profitably use the instrument for real-field applications. We also characterized the single-photon timing resolution, the background noise due to both primary dark count generation and afterpulsing, the single-photon detection efficiency, and the instrument response function shape. The proposed module can become a reliable and cost-effective building block for time-correlated single-photon counting instruments in applications requiring high collection capability of isotropic light and detection efficiency (e.g., fluorescence decay measurements or time-domain diffuse optics systems).« less
NASA Astrophysics Data System (ADS)
Martinenghi, E.; Di Sieno, L.; Contini, D.; Sanzaro, M.; Pifferi, A.; Dalla Mora, A.
2016-07-01
We present the design and preliminary characterization of the first detection module based on Silicon Photomultiplier (SiPM) tailored for single-photon timing applications. The aim of this work is to demonstrate, thanks to the design of a suitable module, the possibility to easily exploit SiPM in many applications as an interesting detector featuring large active area, similarly to photomultipliers tubes, but keeping the advantages of solid state detectors (high quantum efficiency, low cost, compactness, robustness, low bias voltage, and insensitiveness to magnetic field). The module integrates a cooled SiPM with a total photosensitive area of 1 mm2 together with the suitable avalanche signal read-out circuit, the signal conditioning, the biasing electronics, and a Peltier cooler driver for thermal stabilization. It is able to extract the single-photon timing information with resolution better than 100 ps full-width at half maximum. We verified the effective stabilization in response to external thermal perturbations, thus proving the complete insensitivity of the module to environment temperature variations, which represents a fundamental parameter to profitably use the instrument for real-field applications. We also characterized the single-photon timing resolution, the background noise due to both primary dark count generation and afterpulsing, the single-photon detection efficiency, and the instrument response function shape. The proposed module can become a reliable and cost-effective building block for time-correlated single-photon counting instruments in applications requiring high collection capability of isotropic light and detection efficiency (e.g., fluorescence decay measurements or time-domain diffuse optics systems).
Kirchner, Elsa A; Kim, Su Kyoung
2018-01-01
Event-related potentials (ERPs) are often used in brain-computer interfaces (BCIs) for communication or system control for enhancing or regaining control for motor-disabled persons. Especially results from single-trial EEG classification approaches for BCIs support correlations between single-trial ERP detection performance and ERP expression. Hence, BCIs can be considered as a paradigm shift contributing to new methods with strong influence on both neuroscience and clinical applications. Here, we investigate the relevance of the choice of training data and classifier transfer for the interpretability of results from single-trial ERP detection. In our experiments, subjects performed a visual-motor oddball task with motor-task relevant infrequent ( targets ), motor-task irrelevant infrequent ( deviants ), and motor-task irrelevant frequent ( standards ) stimuli. Under dual-task condition, a secondary senso-motor task was performed, compared to the simple-task condition. For evaluation, average ERP analysis and single-trial detection analysis with different numbers of electrodes were performed. Further, classifier transfer was investigated between simple and dual task. Parietal positive ERPs evoked by target stimuli (but not by deviants) were expressed stronger under dual-task condition, which is discussed as an increase of task emphasis and brain processes involved in task coordination and change of task set. Highest classification performance was found for targets irrespective whether all 62, 6 or 2 parietal electrodes were used. Further, higher detection performance of targets compared to standards was achieved under dual-task compared to simple-task condition in case of training on data from 2 parietal electrodes corresponding to results of ERP average analysis. Classifier transfer between tasks improves classification performance in case that training took place on more varying examples (from dual task). In summary, we showed that P300 and overlaying parietal
Single-beam dielectric-microsphere trapping with optical heterodyne detection
NASA Astrophysics Data System (ADS)
Rider, Alexander D.; Blakemore, Charles P.; Gratta, Giorgio; Moore, David C.
2018-01-01
A technique to levitate and measure the three-dimensional position of micrometer-sized dielectric spheres with heterodyne detection is presented. The two radial degrees of freedom are measured by interfering light transmitted through the microsphere with a reference wavefront, while the axial degree of freedom is measured from the phase of the light reflected from the surface of the microsphere. This method pairs the simplicity and accessibility of single-beam optical traps to a measurement of displacement that is intrinsically calibrated by the wavelength of the trapping light and has exceptional immunity to stray light. A theoretical shot noise limit of 1.3 ×10-13 m /√{Hz } for the radial degrees of freedom, and 3.0 ×10-15 m /√{Hz } for the axial degree of freedom can be obtained in the system described. The measured acceleration noise in the radial direction is 7.5 ×10-5 (m /s2) /√{Hz } .
Single- and Dual-Process Models of Biased Contingency Detection
2016-01-01
Abstract. Decades of research in causal and contingency learning show that people’s estimations of the degree of contingency between two events are easily biased by the relative probabilities of those two events. If two events co-occur frequently, then people tend to overestimate the strength of the contingency between them. Traditionally, these biases have been explained in terms of relatively simple single-process models of learning and reasoning. However, more recently some authors have found that these biases do not appear in all dependent variables and have proposed dual-process models to explain these dissociations between variables. In the present paper we review the evidence for dissociations supporting dual-process models and we point out important shortcomings of this literature. Some dissociations seem to be difficult to replicate or poorly generalizable and others can be attributed to methodological artifacts. Overall, we conclude that support for dual-process models of biased contingency detection is scarce and inconclusive. PMID:27025532
Single- and Dual-Process Models of Biased Contingency Detection.
Vadillo, Miguel A; Blanco, Fernando; Yarritu, Ion; Matute, Helena
2016-01-01
Decades of research in causal and contingency learning show that people's estimations of the degree of contingency between two events are easily biased by the relative probabilities of those two events. If two events co-occur frequently, then people tend to overestimate the strength of the contingency between them. Traditionally, these biases have been explained in terms of relatively simple single-process models of learning and reasoning. However, more recently some authors have found that these biases do not appear in all dependent variables and have proposed dual-process models to explain these dissociations between variables. In the present paper we review the evidence for dissociations supporting dual-process models and we point out important shortcomings of this literature. Some dissociations seem to be difficult to replicate or poorly generalizable and others can be attributed to methodological artifacts. Overall, we conclude that support for dual-process models of biased contingency detection is scarce and inconclusive.
Maruyama, Kohei; Takeyama, Haruko; Nemoto, Etsuo; Tanaka, Tsuyoshi; Yoda, Kiyoshi; Matsunaga, Tadashi
2004-09-20
Single nucleotide polymorphism (SNP) detection for aldehyde dehydrogenase 2 (ALDH2) gene based on DNA thermal dissociation curve analysis was successfully demonstrated using an automated system with bacterial magnetic particles (BMPs) by developing a new method for avoiding light scattering caused by nanometer-size particles when using commercially available fluorescent dyes such as FITC, Cy3, and Cy5 as labeling chromophores. Biotin-labeled PCR products in ALDH2, two allele-specific probes (Cy3-labeled detection probe for ALDH2*1 and Cy5-labeled detection probe for ALDH2*2), streptavidin-immobilized BMPs (SA-BMPs) were simultaneously mixed. The mixture was denatured at 70 degrees C for 3 min, cooled slowly to 25 degrees C, and incubated for 10 min, allowing the DNA duplex to form between Cy3- or Cy5-labeled detection probes and biotin-labeled PCR products on SA-BMPs. Then duplex DNA-BMP complex was heated to 58 degrees C, a temperature determined by dissociation curve analysis and a dissociated single-base mismatched detection probe was removed at the same temperature under precise control. Furthermore, fluorescence signal from the detection probe was liberated into the supernatant from completely matched duplex DNA-BMP complex by heating to 80 degrees C and measured. In the homozygote target DNA (ALDH2*1/*1 and ALDH2*2/*2), the fluorescence signals from single-base mismatched were decreased to background level, indicating that mismatched hybridization was efficiently removed by the washing process. In the heterozygote target DNA (ALDH2*1/*2), each fluorescence signals was at a similar level. Therefore, three genotypes of SNP in ALDH2 gene were detected using the automated detection system with BMPs. Copyright 2004 Wiley Periodicals, Inc.
Non-destructive single-pass low-noise detection of ions in a beamline
DOE Office of Scientific and Technical Information (OSTI.GOV)
Schmidt, Stefan; Institut für Kernchemie, Johannes Gutenberg–Universität Mainz, 55099 Mainz; Murböck, Tobias
2015-11-15
We have conceived, built, and operated a device for the non-destructive single-pass detection of charged particles in a beamline. The detector is based on the non-resonant pick-up and subsequent low-noise amplification of the image charges induced in a cylindrical electrode surrounding the particles’ beam path. The first stage of the amplification electronics is designed to be operated from room temperature down to liquid helium temperature. The device represents a non-destructive charge counter as well as a sensitive timing circuit. We present the concept and design details of the device. We have characterized its performance and show measurements with low-energy highlymore » charged ions (such as Ar{sup 13+}) passing through one of the electrodes of a cylindrical Penning trap. This work demonstrates a novel approach of non-destructive, low noise detection of charged particles which is, depending on the bunch structure, suitable, e.g., for ion traps, low-energy beamlines or accelerator transfer sections.« less
All-optical in-depth detection of the acoustic wave emitted by a single gold nanorod
NASA Astrophysics Data System (ADS)
Xu, Feng; Guillet, Yannick; Ravaine, Serge; Audoin, Bertrand
2018-04-01
A single gold nanorod dropped on the surface of a silica substrate is used as a transient optoacoustic source of gigahertz hypersounds. We demonstrate the all-optical detection of the as-generated acoustic wave front propagating in the silica substrate. For this purpose, time-resolved femtosecond pump-probe experiments are performed in a reflection configuration. The fundamental breathing mode of the nanorod is detected at 23 GHz by interferometry, and the longitudinal acoustic wave radiated in the silica substrate is detected by time-resolved Brillouin scattering. By tuning the optical probe wavelength from 750 to 900 nm, hypersounds with wavelengths of 260-315 nm are detected in the silica substrate, with corresponding acoustic frequencies in the range of 19-23 GHz. To confirm the origin of these hypersounds, we theoretically analyze the influence of the acoustic excitation spectrum on the temporal envelope of the transient reflectivity. This analysis proves that the acoustic wave detected in the silica substrate results from the excitation of the breathing mode of the nanorod. These results pave the way for performing local in-depth elastic nanoscopy.
Andrei, Veronica; Sharpe, Erica; Vasilescu, Alina; Andreescu, Silvana
2016-08-15
We report the development and characterization of a disposable single use electrochemical sensor based on the oxidase-like activity of nanoceria particles for the detection of phenolic antioxidants. The use of nanoceria in the sensor design enables oxidation of phenolic compounds, particularly those with ortho-dihydroxybenzene functionality, to their corresponding quinones at the surface of a screen printed carbon electrode. Detection is carried out by electrochemical reduction of the resulting quinone at a low applied potential of -0.1V vs the Ag/AgCl electrode. The sensor was optimized and characterized with respect to particle loading, applied potential, response time, detection limit, linear concentration range and sensitivity. The method enabled rapid detection of common phenolic antioxidants including caffeic acid, gallic acid and quercetin in the µM concentration range, and demonstrated good functionality for the analysis of antioxidant content in several wine samples. The intrinsic oxidase-like activity of nanoceria shows promise as a robust tool for sensitive and cost effective analysis of antioxidants using electrochemical detection. Copyright © 2016 Elsevier B.V. All rights reserved.
Computer simulation of gene detection without PCR by single molecule detection
NASA Astrophysics Data System (ADS)
Davis, Lloyd M.; Williams, John G.; Lamb, Don T.
1999-01-01
Pioneer Hi-Bred is developing a low-cost method for rapid screening of DNA, for use in research on elite crop seed genetics. Unamplified genomic DNA with the requisite base sequence is simultaneously labeled by two different colored fluorescent probes, which hybridize near the selected gene. Dual-channel single molecule detection (SMD) within a flow cell, then provides a sensitive and specific assay for the gene. The technique has been demonstrated using frequency- doubled Nd:YAG laser excitation of two visible-wavelength dyes. A prototype instrument employing infrared fluorophores and laser diodes for excitation has been developed. Here, we report results from a Monte Carlo simulation of the new instrument, in which experimentally determined photophysical parameters for candidate infrared dyes are used for parametric studies of experimental operating conditions. Fluorophore photostability is found to be a key factor in determining the instrument sensitivity. Most infrared dyes have poor photostability, resulting in inefficient SMD. However, the normalized cross-correlation function of the photon signals from each of the two channels can still yield a discernable peak, provided that the concentration of dual- labeled molecules is sufficiently high. Further, for low concentrations, processing of the two photon streams with Gaussian -weighted sliding sum digital filters and selection of simultaneously occurring peaks can also provide a sensitive indicator of the presence of dual-labeled molecules, although accidental coincidences must be considered in the interpretation of results.
NASA Astrophysics Data System (ADS)
Singh, Swati; Kumar, Ashok; Khare, Shashi; Mulchandani, Ashok; Rajesh
2014-11-01
A specific and ultrasensitive, label free single-walled carbon nanotubes (SWNTs) based chemiresistive genosensor was fabricated for the early detection of Streptococcus pyogenes infection in human causing rheumatic heart disease. The mga gene of S. pyogenes specific 24 mer ssDNA probe was covalently immobilized on SWNT through a molecular bilinker, 1-pyrenemethylamine, using carbodiimide coupling reaction. The sensor was characterized by the current-voltage (I-V) characteristic curve and scanning electron microscopy. The sensing performance of the sensor was studied with respect to changes in conductance in SWNT channel based on hybridization of the target S. pyogenes single stranded genomic DNA (ssG-DNA) to its complementary 24 mer ssDNA probe. The sensor shows negligible response to non-complementary Staphylococcus aureus ssG-DNA, confirming the specificity of the sensor only with S. pyogenes. The genosensor exhibited a linear response to S. pyogenes G-DNA from 1 to1000 ng ml-1 with a limit of detection of 0.16 ng ml-1.
Host Immunity via Mutable Virtualized Large-Scale Network Containers
2016-07-25
and constrain the distributed persistent inside crawlers that have va.lid credentials to access the web services. The main idea is to add a marker...to each web page URL and use the URL path and user inforn1ation contained in the marker to help accurately detect crawlers at its earliest stage...more than half of all website traffic, and malicious bots contributes almost one third of the traffic. As one type of bots, web crawlers have been
Dendrimer enriched single-use aptasensor for impedimetric detection of activated protein C.
Erdem, Arzum; Congur, Gulsah
2014-05-01
A novel impedimetric aptasensor for detection of human activated protein C (APC) was introduced for the first time in the present study. An enhanced sensor response was obtained using poly(amidoamine) (PAMAM) dendrimer having 16 succinamic acid surface groups (generation 2, G2-PS), that was modified onto the surface of screen printed graphite electrode (G2-PS/SPE). An amino modified DNA aptamer was then immobilized onto the surface of G2-PS modified SPE. The selective interaction of APT with its cognate protein, APC was investigated using different electrochemical techniques; differential pulse voltammetry (DPV), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The microscopic characterization was consecutively performed before/after each modification/interaction step using scanning electron microscopy (SEM) and atomic force microscopy (AFM). The selectivity of aptasensor was tested in the presence of numerous proteins; protein C, thrombin, bovine serum albumin, factor Va and chromogenic substrate in different buffer mediums. The APC detection in the artificial serum; fetal bovine serum (FBS) was also performed impedimetrically. This dendrimer modified aptasensor technology brings several advantages: being single-use, fast screening with low-cost per measurement and resulting in sensitive detection of APC with the detection limits of 0.74 μg/mL (0.46 pmol in 35 μL sample) in buffer medium, and 2.03 μg/mL (1.27 pmol in 35 μL sample) in serum. Copyright © 2014 Elsevier B.V. All rights reserved.
Fiedler, Lorenz; Wöstmann, Malte; Graversen, Carina; Brandmeyer, Alex; Lunner, Thomas; Obleser, Jonas
2017-06-01
Conventional, multi-channel scalp electroencephalography (EEG) allows the identification of the attended speaker in concurrent-listening ('cocktail party') scenarios. This implies that EEG might provide valuable information to complement hearing aids with some form of EEG and to install a level of neuro-feedback. To investigate whether a listener's attentional focus can be detected from single-channel hearing-aid-compatible EEG configurations, we recorded EEG from three electrodes inside the ear canal ('in-Ear-EEG') and additionally from 64 electrodes on the scalp. In two different, concurrent listening tasks, participants (n = 7) were fitted with individualized in-Ear-EEG pieces and were either asked to attend to one of two dichotically-presented, concurrent tone streams or to one of two diotically-presented, concurrent audiobooks. A forward encoding model was trained to predict the EEG response at single EEG channels. Each individual participants' attentional focus could be detected from single-channel EEG response recorded from short-distance configurations consisting only of a single in-Ear-EEG electrode and an adjacent scalp-EEG electrode. The differences in neural responses to attended and ignored stimuli were consistent in morphology (i.e. polarity and latency of components) across subjects. In sum, our findings show that the EEG response from a single-channel, hearing-aid-compatible configuration provides valuable information to identify a listener's focus of attention.
Detection of single electron spin resonance in a double quantum dota)
NASA Astrophysics Data System (ADS)
Koppens, F. H. L.; Buizert, C.; Vink, I. T.; Nowack, K. C.; Meunier, T.; Kouwenhoven, L. P.; Vandersypen, L. M. K.
2007-04-01
Spin-dependent transport measurements through a double quantum dot are a valuable tool for detecting both the coherent evolution of the spin state of a single electron, as well as the hybridization of two-electron spin states. In this article, we discuss a model that describes the transport cycle in this regime, including the effects of an oscillating magnetic field (causing electron spin resonance) and the effective nuclear fields on the spin states in the two dots. We numerically calculate the current flow due to the induced spin flips via electron spin resonance, and we study the detector efficiency for a range of parameters. The experimental data are compared with the model and we find a reasonable agreement.
Yan, Hainian; Yagyu, Fumihiro; Okitsu, Shoko; Nishio, Osamu; Ushijima, Hiroshi
2003-12-01
A reverse transcription (RT) single-round multiplex polymerase chain reaction (smPCR) assay was developed to detect simultaneously Norovirus genogroup I and II, Sapovirus and astrovirus. A total of 377 diarrhea stool samples (screened for rotavirus- and adenorivus-negative) from four regions in Japan during July 2000 to June 2001 were examined by RT-smPCR. The positive rate was 16.4% (62 out of 377 stool samples). Norovirus, Sapovirus and astrovirus were detected in 42, 16, 4 of 60 positive samples, respectively. Coinfection was not found in these samples. Infections occurred mainly in November, December and January. The key elements of the RT-smPCR are (i) the cDNA synthesis with the Superscript RTII and random primer at 42 degrees C for 1 h, at 99 degrees C for 5 min, and (ii) single-round multiplex PCR by using Taq polymerase mixed together with a mixture of four different primer pairs (G1-SKF/G1-SKR for Norovirus genogroup I, COG2F/G2-SKR for Norovirus genogroup II, SLV5317/SLV5749 for Sapovirus, PreCAP1/82b for astrovirus). All of the four primer pairs amplify the capsid region of target viral genome, produce four size-specific amplicons of 330, 387, 434, 719 bp for Norovirus genogroup I and II, Sapovirus and astrovirus, respectively. This assay provides a more rapid and efficient way to detect these viruses from fecal samples in a single test, and also offers the potential for their molecular detection in food and environmental samples.
Detection and direction discrimination of single vortex rings by harbour seals (Phoca vitulina).
Krüger, Yvonne; Hanke, Wolf; Miersch, Lars; Dehnhardt, Guido
2018-04-25
Harbour seals possess highly sensitive vibrissae that enable them to track hydrodynamic trails left behind by a swimming fish. Most of these trails contain vortex rings as a main hydrodynamic component. They may reveal information about their generator as the trails differ depending on the fish species, the fish's body shape, size and swimming style. In addition, fish generate single vortex rings in diverse natural situations. In this study, the ability of blindfolded stationary harbour seals to detect and analyse single vortex rings regarding directional information has been investigated. In three different behavioural experiments, the animals were trained to respond to single artificially generated vortex rings. The results show that harbour seals are able to respond to a variety of different vortex rings upon vibrissal stimulation. The investigation of the minimum hydrodynamically perceivable angle revealed that it is at least as small as 5.7 deg, which was the smallest adjustable angle. Moreover, harbour seals are capable of analysing the travel direction of a vortex ring perceived by the mystacial vibrissae irrespective of whether the vibrissae were stimulated ipsilaterally or contralaterally. In situations in which no complex hydrodynamic trail is available, it is advantageous for a hunting seal to be able to extract information from a single vortex ring. © 2018. Published by The Company of Biologists Ltd.
Analysis of hybrid subcarrier multiplexing of OCDMA based on single photodiode detection
NASA Astrophysics Data System (ADS)
Ahmad, N. A. A.; Junita, M. N.; Aljunid, S. A.; Rashidi, C. B. M.; Endut, R.
2017-11-01
This paper analyzes the performance of subcarrier multiplexing (SCM) of spectral amplitude coding optical code multiple access (SAC-OCDMA) by applying Recursive Combinatorial (RC) code based on single photodiode detection (SPD). SPD is used in the receiver part to reduce the effect of multiple access interference (MAI) which contributes as a dominant noise in incoherent SAC-OCDMA systems. Results indicate that the SCM OCDMA network performance could be improved by using lower data rates and higher number of weight. Total number of users can also be enhanced by adding lower data rates and higher number of subcarriers.
Detecting uniaxial single domain grains with a modified IRM technique
NASA Astrophysics Data System (ADS)
Mitra, R.; Tauxe, L.; Gee, J. S.
2011-12-01
Mid-ocean ridge basalt (MORB) specimens have often been found to have high ratios of saturation remanence to saturation magnetization (Mrs/Ms). This has been attributed either to dominant cubic anisotropy or to insufficient saturating field leading to overestimation of Mrs/Ms of a dominantly uniaxial single domain (USD) assemblage. To resolve this debate, we develop an independent technique to detect USD assemblages. The experimental protocol involves subjecting the specimen to bidirectional impulse fields at each step. The experiment is similar to the conventional isothermal remanent magnetization (IRM) acquisition experiment but the field is applied twice, in antiparallel directions. We define a new parameter, IRAT, as the ratio of the remanences at each field step and show it to have characteristic behaviour for the two assemblages; IRAT ˜1 at all field steps for USD and <1 with a strong field dependence for multi-axial single domain (MSD) grains. We verified the theoretical predictions experimentally with representative USD and MSD specimens. Experiments with MORBs gave low IRATs for specimens having high Mrs/Ms. This argues for a dominant MSD assemblage in the MORBs, possibly cubic in nature. Although undersaturation of the samples can indeed be a contributing factor to the exceptionally high Mrs/Ms, this study shows that the nature of the assemblage cannot be dominantly USD.
Wood-Bouwens, Christina; Lau, Billy T; Handy, Christine M; Lee, HoJoon; Ji, Hanlee P
2017-09-01
We describe a single-color digital PCR assay that detects and quantifies cancer mutations directly from circulating DNA collected from the plasma of cancer patients. This approach relies on a double-stranded DNA intercalator dye and paired allele-specific DNA primer sets to determine an absolute count of both the mutation and wild-type-bearing DNA molecules present in the sample. The cell-free DNA assay uses an input of 1 ng of nonamplified DNA, approximately 300 genome equivalents, and has a molecular limit of detection of three mutation DNA genome-equivalent molecules per assay reaction. When using more genome equivalents as input, we demonstrated a sensitivity of 0.10% for detecting the BRAF V600E and KRAS G12D mutations. We developed several mutation assays specific to the cancer driver mutations of patients' tumors and detected these same mutations directly from the nonamplified, circulating cell-free DNA. This rapid and high-performance digital PCR assay can be configured to detect specific cancer mutations unique to an individual cancer, making it a potentially valuable method for patient-specific longitudinal monitoring. Copyright © 2017 American Society for Investigative Pathology and the Association for Molecular Pathology. Published by Elsevier Inc. All rights reserved.
Single-photon technique for the detection of periodic extraterrestrial laser pulses.
Leeb, W R; Poppe, A; Hammel, E; Alves, J; Brunner, M; Meingast, S
2013-06-01
To draw humankind's attention to its existence, an extraterrestrial civilization could well direct periodic laser pulses toward Earth. We developed a technique capable of detecting a quasi-periodic light signal with an average of less than one photon per pulse within a measurement time of a few tens of milliseconds in the presence of the radiation emitted by an exoplanet's host star. Each of the electronic events produced by one or more single-photon avalanche detectors is tagged with precise time-of-arrival information and stored. From this we compute a histogram displaying the frequency of event-time differences in classes with bin widths on the order of a nanosecond. The existence of periodic laser pulses manifests itself in histogram peaks regularly spaced at multiples of the-a priori unknown-pulse repetition frequency. With laser sources simulating both the pulse source and the background radiation, we tested a detection system in the laboratory at a wavelength of 850 nm. We present histograms obtained from various recorded data sequences with the number of photons per pulse, the background photons per pulse period, and the recording time as main parameters. We then simulated a periodic signal hypothetically generated on a planet orbiting a G2V-type star (distance to Earth 500 light-years) and show that the technique is capable of detecting the signal even if the received pulses carry as little as one photon on average on top of the star's background light.
Lapitan Jr., Lorico D. S.; Guo, Yuan
2015-01-01
Single nucleotide polymorphisms (SNPs) constitute the most common types of genetic variations in the human genome. A number of SNPs have been linked to the development of life threatening diseases including cancer, cardiovascular diseases and neurodegenerative diseases. The ability for ultrasensitive and accurate detection of low abundant disease-related SNPs in bodily fluids (e.g. blood, serum, etc.) holds a significant value in the development of non-invasive future biodiagnostic tools. Over the past two decades, nanomaterials have been utilized in a myriad of biosensing applications due to their ability of detecting extremely low quantities of biologically important biomarkers with high sensitivity and accuracy. Of particular interest is the application of such technologies in the detection of SNPs. The use of various nanomaterials, coupled with different powerful signal amplification strategies, has paved the way for a new generation of ultrasensitive SNP biodiagnostic assays. Over the past few years, several ultrasensitive SNP biosensors capable of detecting specific targets down to the ultra-low regimes (ca. aM and below) and therefore holding great promises for early clinical diagnosis of diseases have been developed. This mini review will highlight some of the most recent, significant advances in nanomaterial-based ultrasensitive SNP sensing technologies capable of detecting specific targets on the attomolar (10–18 M) regime or below. In particular, the design of novel, powerful signal amplification strategies that hold the key to the ultrasensitivity is highlighted. PMID:25785914
Urtnasan, Erdenebayar; Park, Jong-Uk; Joo, Eun-Yeon; Lee, Kyoung-Joung
2018-04-23
In this study, we propose a method for the automated detection of obstructive sleep apnea (OSA) from a single-lead electrocardiogram (ECG) using a convolutional neural network (CNN). A CNN model was designed with six optimized convolution layers including activation, pooling, and dropout layers. One-dimensional (1D) convolution, rectified linear units (ReLU), and max pooling were applied to the convolution, activation, and pooling layers, respectively. For training and evaluation of the CNN model, a single-lead ECG dataset was collected from 82 subjects with OSA and was divided into training (including data from 63 patients with 34,281 events) and testing (including data from 19 patients with 8571 events) datasets. Using this CNN model, a precision of 0.99%, a recall of 0.99%, and an F 1 -score of 0.99% were attained with the training dataset; these values were all 0.96% when the CNN was applied to the testing dataset. These results show that the proposed CNN model can be used to detect OSA accurately on the basis of a single-lead ECG. Ultimately, this CNN model may be used as a screening tool for those suspected to suffer from OSA.
Detection and capture of single circulating melanoma cells using photoacoustic flowmetry
NASA Astrophysics Data System (ADS)
O'Brien, Christine; Mosley, Jeffrey; Goldschmidt, Benjamin S.; Viator, John A.
2010-02-01
Photoacoustic flowmetry has been used to detect single circulating melanoma cells in vitro. Circulating melanoma cells are those cells that travel in the blood and lymph systems to create secondary tumors and are the hallmark of metastasis. This technique involves taking blood samples from patients, separating the white blood and melanoma cells from whole blood and irradiating them with a pulsed laser in a flowmetry set up. Rapid, visible wavelength laser pulses on the order of 5 ns can induce photoacoustic waves in melanoma cells due to their melanin content, while surrounding white blood cells remain acoustically passive. We have developed a system that identifies rare melanoma cells and captures them in 50 microliter volumes using suction applied near the photoacoustic detection chamber. The 50 microliter sample is then diluted and the experiment is repeated using the new sample until only a melanoma cell remains. We have tested this system on dyed microspheres ranging in size from 300 to 500 microns. Capture of circulating melanoma cells may provide the opportunity to study metastatic cells for basic understanding of the spread of cancer and to optimize patient specific therapies.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Gust, I.D.; Feinstone, S.M.; Purcell, R.H.
1980-01-01
A sensitive ''Farr'' assay, utilizing /sup 125/I-labelled DNA was developed for detecting antibody to single-stranded DNA (anti-ssDNA). The test was shown to be specific and as sensitive as assays using /sup 14/C-labelled DNA, for the detection of antibody in patients with connective tissue diseases. Groups of sera from patients with naturally acquired viral hepatitis and experimentally infected chimpanzees were tested for anti-ssDNA by the /sup 125/I assay and by counterimmunoelectrophoresis (CIEP). No consistent pattern was observed with either technique, indicating the elevated levels of this antibody are not as reliable markers of parenchymal liver damage as had been previously suggested.
Katz, Ben; Minke, Baruch
2012-01-01
Drosophila photoreceptor cells use the ubiquitous G-protein-mediated phospholipase C (PLC) cascade to achieve ultimate single photon sensitivity. This is manifested in the single photon responses (quantum bumps). In photoreceptor cells, dark activation of Gqα molecules occurs spontaneously and produces unitary dark events (dark bumps). A high rate of spontaneous Gqα activation and dark bump production potentially hampers single photon detection. We found that in wild type flies the in vivo rate of spontaneous Gqα activation is very high. Nevertheless, this high rate is not manifested in a substantially high rate of dark bumps. Therefore, it is unclear how phototransduction suppresses dark bump production, arising from spontaneous Gqα activation, while still maintaining high-fidelity representation of single photons. In this study we show that reduced PLC catalytic activity selectively suppressed production of dark bumps but not light-induced bumps. Manipulations of PLC activity using PLC mutant flies and Ca2+ modulations revealed that a critical level of PLC activity is required to induce bump production. The required minimal level of PLC activity, selectively suppressed random production of single Gqα-activated dark bumps despite a high rate of spontaneous Gqα activation. This minimal PLC activity level is reliably obtained by photon induced synchronized activation of several neighboring Gqα molecules activating several PLC molecules, but not by random activation of single Gqα molecules. We thus demonstrate how a G-protein-mediated transduction system, with PLC as its target, selectively suppresses its intrinsic noise while preserving reliable signaling. PMID:22357856
Ardley, Nicholas D; Lau, Ken K; Buchan, Kevin
2013-12-01
Cervical spine injuries occur in 4-8 % of adults with head trauma. Dual acquisition technique has been traditionally used for the CT scanning of brain and cervical spine. The purpose of this study was to determine the efficacy of radiation dose reduction by using a single acquisition technique that incorporated both anatomical regions with a dedicated neck detection algorithm. Thirty trauma patients for brain and cervical spine CT were included and were scanned with the single acquisition technique. The radiation doses from the single CT acquisition technique with the neck detection algorithm, which allowed appropriate independent dose administration relevant to brain and cervical spine regions, were recorded. Comparison was made both to the doses calculated from the simulation of the traditional dual acquisitions with matching parameters, and to the doses of retrospective dual acquisition legacy technique with the same sample size. The mean simulated dose for the traditional dual acquisition technique was 3.99 mSv, comparable to the average dose of 4.2 mSv from 30 previous patients who had CT of brain and cervical spine as dual acquisitions. The mean dose from the single acquisition technique was 3.35 mSv, resulting in a 16 % overall dose reduction. The images from the single acquisition technique were of excellent diagnostic quality. The new single acquisition CT technique incorporating the neck detection algorithm for brain and cervical spine significantly reduces the overall radiation dose by eliminating the unavoidable overlapping range between 2 anatomical regions which occurs with the traditional dual acquisition technique.
NASA Astrophysics Data System (ADS)
Luo, Qingying; Liu, Lin; Yang, Cai; Yuan, Jing; Feng, Hongtao; Chen, Yan; Zhao, Peng; Yu, Zhiqiang; Jin, Zongwen
2018-03-01
MicroRNAs (miRNAs) are single stranded endogenous molecules composed of only 18-24 nucleotides which are critical for gene expression regulating the translation of messenger RNAs. Conventional methods based on enzyme-assisted nucleic acid amplification techniques have many problems, such as easy contamination, high cost, susceptibility to false amplification, and tendency to have sequence mismatches. Here we report a rapid, ratiometric, enzyme-free, sensitive, and highly selective single-step miRNA detection using three-way junction assembled (or self-assembled) FRET probes. The developed strategy can be operated within the linear range from subnanomolar to hundred nanomolar concentrations of miRNAs. In comparison with the traditional approaches, our method showed high sensitivity for the miRNA detection and extreme selectivity for the efficient discrimination of single-base mismatches. The results reveal that the strategy paved a new avenue for the design of novel highly specific probes applicable in diagnostics and potentially in microscopic imaging of miRNAs in real biological environments.
NASA Astrophysics Data System (ADS)
Fiedler, Lorenz; Wöstmann, Malte; Graversen, Carina; Brandmeyer, Alex; Lunner, Thomas; Obleser, Jonas
2017-06-01
Objective. Conventional, multi-channel scalp electroencephalography (EEG) allows the identification of the attended speaker in concurrent-listening (‘cocktail party’) scenarios. This implies that EEG might provide valuable information to complement hearing aids with some form of EEG and to install a level of neuro-feedback. Approach. To investigate whether a listener’s attentional focus can be detected from single-channel hearing-aid-compatible EEG configurations, we recorded EEG from three electrodes inside the ear canal (‘in-Ear-EEG’) and additionally from 64 electrodes on the scalp. In two different, concurrent listening tasks, participants (n = 7) were fitted with individualized in-Ear-EEG pieces and were either asked to attend to one of two dichotically-presented, concurrent tone streams or to one of two diotically-presented, concurrent audiobooks. A forward encoding model was trained to predict the EEG response at single EEG channels. Main results. Each individual participants’ attentional focus could be detected from single-channel EEG response recorded from short-distance configurations consisting only of a single in-Ear-EEG electrode and an adjacent scalp-EEG electrode. The differences in neural responses to attended and ignored stimuli were consistent in morphology (i.e. polarity and latency of components) across subjects. Significance. In sum, our findings show that the EEG response from a single-channel, hearing-aid-compatible configuration provides valuable information to identify a listener’s focus of attention.
Germline contamination and leakage in whole genome somatic single nucleotide variant detection.
Sendorek, Dorota H; Caloian, Cristian; Ellrott, Kyle; Bare, J Christopher; Yamaguchi, Takafumi N; Ewing, Adam D; Houlahan, Kathleen E; Norman, Thea C; Margolin, Adam A; Stuart, Joshua M; Boutros, Paul C
2018-01-31
The clinical sequencing of cancer genomes to personalize therapy is becoming routine across the world. However, concerns over patient re-identification from these data lead to questions about how tightly access should be controlled. It is not thought to be possible to re-identify patients from somatic variant data. However, somatic variant detection pipelines can mistakenly identify germline variants as somatic ones, a process called "germline leakage". The rate of germline leakage across different somatic variant detection pipelines is not well-understood, and it is uncertain whether or not somatic variant calls should be considered re-identifiable. To fill this gap, we quantified germline leakage across 259 sets of whole-genome somatic single nucleotide variant (SNVs) predictions made by 21 teams as part of the ICGC-TCGA DREAM Somatic Mutation Calling Challenge. The median somatic SNV prediction set contained 4325 somatic SNVs and leaked one germline polymorphism. The level of germline leakage was inversely correlated with somatic SNV prediction accuracy and positively correlated with the amount of infiltrating normal cells. The specific germline variants leaked differed by tumour and algorithm. To aid in quantitation and correction of leakage, we created a tool, called GermlineFilter, for use in public-facing somatic SNV databases. The potential for patient re-identification from leaked germline variants in somatic SNV predictions has led to divergent open data access policies, based on different assessments of the risks. Indeed, a single, well-publicized re-identification event could reshape public perceptions of the values of genomic data sharing. We find that modern somatic SNV prediction pipelines have low germline-leakage rates, which can be further reduced, especially for cloud-sharing, using pre-filtering software.
Multiplexed Detection of Cytokines Based on Dual Bar-Code Strategy and Single-Molecule Counting.
Li, Wei; Jiang, Wei; Dai, Shuang; Wang, Lei
2016-02-02
Cytokines play important roles in the immune system and have been regarded as biomarkers. While single cytokine is not specific and accurate enough to meet the strict diagnosis in practice, in this work, we constructed a multiplexed detection method for cytokines based on dual bar-code strategy and single-molecule counting. Taking interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) as model analytes, first, the magnetic nanobead was functionalized with the second antibody and primary bar-code strands, forming a magnetic nanoprobe. Then, through the specific reaction of the second antibody and the antigen that fixed by the primary antibody, sandwich-type immunocomplex was formed on the substrate. Next, the primary bar-code strands as amplification units triggered multibranched hybridization chain reaction (mHCR), producing nicked double-stranded polymers with multiple branched arms, which were served as secondary bar-code strands. Finally, the secondary bar-code strands hybridized with the multimolecule labeled fluorescence probes, generating enhanced fluorescence signals. The numbers of fluorescence dots were counted one by one for quantification with epi-fluorescence microscope. By integrating the primary and secondary bar-code-based amplification strategy and the multimolecule labeled fluorescence probes, this method displayed an excellent sensitivity with the detection limits were both 5 fM. Unlike the typical bar-code assay that the bar-code strands should be released and identified on a microarray, this method is more direct. Moreover, because of the selective immune reaction and the dual bar-code mechanism, the resulting method could detect the two targets simultaneously. Multiple analysis in human serum was also performed, suggesting that our strategy was reliable and had a great potential application in early clinical diagnosis.
NASA Astrophysics Data System (ADS)
Nygren, David
2015-10-01
To proceed toward effective ``discovery class'' ton-scale detectors in the search for neutrino-less double beta decay, a robust technique for rejection of all radioactivity-induced backgrounds is urgently needed. An efficient technique for detection of the barium daughter in the decay 136Xe -->136Ba + 2e- would provide a long-sought pathway toward this goal. Single-molecule fluorescent imaging appears to offer a new way to detect the barium daughter atom, which emerges naturally in an ionized state in pure xenon. A doubly charged barium ion can initiate a chelation process with a non-fluorescent precursor molecule, leading to a highly fluorescent complex. Repeated photo-excitation of the complex can reveal both presence and location of a single ionized atom with high precision and selectivity. Detection within the active volume of a xenon gas Time Projection Chamber operating at high pressure would be automatic, and with a capability for redundant confirmation.
A New Single-Step PCR Assay for the Detection of the Zoonotic Malaria Parasite Plasmodium knowlesi
Lucchi, Naomi W.; Poorak, Mitra; Oberstaller, Jenna; DeBarry, Jeremy; Srinivasamoorthy, Ganesh; Goldman, Ira; Xayavong, Maniphet; da Silva, Alexandre J.; Peterson, David S.; Barnwell, John W.; Kissinger, Jessica; Udhayakumar, Venkatachalam
2012-01-01
Background Recent studies in Southeast Asia have demonstrated substantial zoonotic transmission of Plasmodium knowlesi to humans. Microscopically, P. knowlesi exhibits several stage-dependent morphological similarities to P. malariae and P. falciparum. These similarities often lead to misdiagnosis of P. knowlesi as either P. malariae or P. falciparum and PCR-based molecular diagnostic tests are required to accurately detect P. knowlesi in humans. The most commonly used PCR test has been found to give false positive results, especially with a proportion of P. vivax isolates. To address the need for more sensitive and specific diagnostic tests for the accurate diagnosis of P. knowlesi, we report development of a new single-step PCR assay that uses novel genomic targets to accurately detect this infection. Methodology and Significant Findings We have developed a bioinformatics approach to search the available malaria parasite genome database for the identification of suitable DNA sequences relevant for molecular diagnostic tests. Using this approach, we have identified multi-copy DNA sequences distributed in the P. knowlesi genome. We designed and tested several novel primers specific to new target sequences in a single-tube, non-nested PCR assay and identified one set of primers that accurately detects P. knowlesi. We show that this primer set has 100% specificity for the detection of P. knowlesi using three different strains (Nuri, H, and Hackeri), and one human case of malaria caused by P. knowlesi. This test did not show cross reactivity with any of the four human malaria parasite species including 11 different strains of P. vivax as well as 5 additional species of simian malaria parasites. Conclusions The new PCR assay based on novel P. knowlesi genomic sequence targets was able to accurately detect P. knowlesi. Additional laboratory and field-based testing of this assay will be necessary to further validate its utility for clinical diagnosis of P. knowlesi. PMID
Zhu, Shuyun; Liu, Zhongyuan; Hu, Lianzhe; Yuan, Yali; Xu, Guobao
2012-12-14
Proteases play a central role in several widespread diseases. Thus, there is a great need for the fast and sensitive detection of various proteolytic enzymes. Herein, we have developed a carbon nanotube (CNT)-based protease biosensing platform that uses peptides as a fluorescence probe for the first time. Single-walled carbon nanohorns (SWCNHs) and thrombin were used to demonstrate this detection strategy. SWCNHs can adsorb a fluorescein-based dye (FAM)-labeled peptide (FAM-pep) and quench the fluorescence of FAM. In contrast, thrombin can cleave FAM-pep on SWCNHs and recover the fluorescence of FAM, which allows the sensitive detection of thrombin. This biosensor has a high sensitivity and selectivity toward thrombin, with a detection limit of 100 pM. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Agarwalla, Bijay Kumar; Hua, Weijie; Zhang, Yu
2015-06-07
The nonlinear optical response of a current-carrying single molecule coupled to two metal leads and driven by a sequence of impulsive optical pulses with controllable phases and time delays is calculated. Coherent (stimulated, heterodyne) detection of photons and incoherent detection of the optically induced current are compared. Using a diagrammatic Liouville space superoperator formalism, the signals are recast in terms of molecular correlation functions which are then expanded in the many-body molecular states. Two dimensional signals in benzene-1,4-dithiol molecule show cross peaks involving charged states. The correlation between optical and charge current signal is also observed.
Gonnissen, J; De Backer, A; den Dekker, A J; Sijbers, J; Van Aert, S
2016-11-01
In the present paper, the optimal detector design is investigated for both detecting and locating light atoms from high resolution scanning transmission electron microscopy (HR STEM) images. The principles of detection theory are used to quantify the probability of error for the detection of light atoms from HR STEM images. To determine the optimal experiment design for locating light atoms, use is made of the so-called Cramér-Rao Lower Bound (CRLB). It is investigated if a single optimal design can be found for both the detection and location problem of light atoms. Furthermore, the incoming electron dose is optimised for both research goals and it is shown that picometre range precision is feasible for the estimation of the atom positions when using an appropriate incoming electron dose under the optimal detector settings to detect light atoms. Copyright © 2016 Elsevier B.V. All rights reserved.
Wu, Wei; Chen, Junhua; Fang, Zhiyuan; Ge, Chenchen; Xiang, Zhicheng; Ouyang, Chuanyan; Lie, Puchang; Xiao, Zhuo; Yu, Luxin; Wang, Lin; Zeng, Lingwen
2013-12-04
Polymerase-free and label-free strategies for DNA detection have shown excellent sensitivity and specificity in various biological samples. Herein, we propose a method for single nucleotide polymorphism (SNP) detection by using self-assembled DNA concatemers. Capture probes, bound to magnetic beads, can joint mediator probes by T4 DNA ligase in the presence of target DNA that is complementary to the capture probe and mediator probe. The mediator probes trigger self-assembly of two auxiliary probes on magnetic beads to form DNA concatemers. Separated by a magnetic rack, the double-stranded concatemers on beads can recruit a great amount of SYBR Green I and eventually result in amplified fluorescent signals. In comparison with reported methods for SNP detection, the concatemer-based approach has significant advantages of low background, simplicity, and ultrasensitivity, making it as a convenient platform for clinical applications. As a proof of concept, BRAF(T1799A) oncogene mutation, a SNP involved in diverse human cancers, was used as a model target. The developed approach using a fluorescent intercalator can detect as low as 0.1 fM target BRAF(T1799A) DNA, which is better than those previously published methods for SNP detection. This method is robust and can be used directly to measure the BRAF(T1799A) DNA in complex human serum with excellent recovery (94-103%). It is expected that this assay principle can be directed toward other SNP genes by simply changing the mediator probe and auxiliary probes. Copyright © 2013 Elsevier B.V. All rights reserved.
Optical tweezers with 2.5 kHz bandwidth video detection for single-colloid electrophoresis
NASA Astrophysics Data System (ADS)
Otto, Oliver; Gutsche, Christof; Kremer, Friedrich; Keyser, Ulrich F.
2008-02-01
We developed an optical tweezers setup to study the electrophoretic motion of colloids in an external electric field. The setup is based on standard components for illumination and video detection. Our video based optical tracking of the colloid motion has a time resolution of 0.2ms, resulting in a bandwidth of 2.5kHz. This enables calibration of the optical tweezers by Brownian motion without applying a quadrant photodetector. We demonstrate that our system has a spatial resolution of 0.5nm and a force sensitivity of 20fN using a Fourier algorithm to detect periodic oscillations of the trapped colloid caused by an external ac field. The electrophoretic mobility and zeta potential of a single colloid can be extracted in aqueous solution avoiding screening effects common for usual bulk measurements.
Ivanov, A N; Younusov, R R; Evtugyn, G A; Arduini, F; Moscone, D; Palleschi, G
2011-07-15
A simple and reliable technique has been developed for the construction of an amperometric acetylcholinesterase biosensor based on screen-printed carbon electrodes. For the first time, one-step modification using single-walled carbon nanotubes and Co phtalocyanine has been proposed to decrease the working potential and to increase the signal of thiocholine oxidation. The biosensor developed made it possible to detect 5-50 ppb of paraoxon and 2-50 ppb of malaoxon with detection limits of 3 and 2 ppb, respectively (incubation 15 min). The biosensor showed high reproducibility when measurements of the substrate and inhibitor were performed (R.S.D. about 1% and 2.5%, respectively). The reliability of the inhibition measurements was confirmed by testing spiked samples of sparkling and tape waters. Copyright © 2011 Elsevier B.V. All rights reserved.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Singh, Swati; Kumar, Ashok, E-mail: rajesh-csir@yahoo.com, E-mail: ashokigib@rediffmail.com; Academy of Scientific and Innovative Research
A specific and ultrasensitive, label free single-walled carbon nanotubes (SWNTs) based chemiresistive genosensor was fabricated for the early detection of Streptococcus pyogenes infection in human causing rheumatic heart disease. The mga gene of S. pyogenes specific 24 mer ssDNA probe was covalently immobilized on SWNT through a molecular bilinker, 1-pyrenemethylamine, using carbodiimide coupling reaction. The sensor was characterized by the current-voltage (I-V) characteristic curve and scanning electron microscopy. The sensing performance of the sensor was studied with respect to changes in conductance in SWNT channel based on hybridization of the target S. pyogenes single stranded genomic DNA (ssG-DNA) to itsmore » complementary 24 mer ssDNA probe. The sensor shows negligible response to non-complementary Staphylococcus aureus ssG-DNA, confirming the specificity of the sensor only with S. pyogenes. The genosensor exhibited a linear response to S. pyogenes G-DNA from 1 to1000 ng ml{sup −1} with a limit of detection of 0.16 ng ml{sup −1}.« less
Epithelial borderline ovarian tumor: Diagnosis and treatment strategy.
Ushijima, Kimio; Kawano, Kouichiro; Tsuda, Naotake; Nishio, Shin; Terada, Atsumu; Kato, Hiroyuki; Tasaki, Kazuto; Matsukuma, Ken
2015-05-01
Epithelial borderline ovarian tumors (BOT) are distinctive from benign tumors and carcinoma. They occur in younger women more often than carcinoma, and there is some difficulty making correct diagnosis of BOT. Two subtypes of BOT, serous and mucinous borderline tumor have different characteristics and very different clinical behavior. Serous borderline tumor (SBT) with micropapillary pattern shows more incidence of extra ovarian disease and often coexists with invasive implant. SBT with micropapillary pattern in advanced stage has showed a worse prognosis than typical SBT. Huge mucinous borderline tumors have histologic heterogeneity, and the accuracy of frozen section diagnosis is relatively low. Extensive sampling is required to reach a correct pathological diagnosis. Mucinous adenoma (intestinal type) also runs the risk of recurrence after cystectomy, or intraoperative rupture of cyst. Laparoscopic procedure for BOT has not increased the risk of recurrence. Fertility preserving procedures are generally accepted, except in advanced stage SBT with invasive implants. Only cystectomy shows a significant risk of recurrence. Re-staging surgery and full staging surgery is not necessary for all BOT. We should not attempt to treat them uniformly, by the single diagnosis of "borderline tumor". It depends on histologic type. Close communication with the pathologist is necessary to gain more detail and ask more pathological samples in order to make the optimal treatment strategy for each individual patients.
Measuring temporal summation in visual detection with a single-photon source.
Holmes, Rebecca; Victora, Michelle; Wang, Ranxiao Frances; Kwiat, Paul G
2017-11-01
Temporal summation is an important feature of the visual system which combines visual signals that arrive at different times. Previous research estimated complete summation to last for 100ms for stimuli judged "just detectable." We measured the full range of temporal summation for much weaker stimuli using a new paradigm and a novel light source, developed in the field of quantum optics for generating small numbers of photons with precise timing characteristics and reduced variance in photon number. Dark-adapted participants judged whether a light was presented to the left or right of their fixation in each trial. In Experiment 1, stimuli contained a stream of photons delivered at a constant rate while the duration was systematically varied. Accuracy should increase with duration as long as the later photons can be integrated with the proceeding ones into a single signal. The temporal integration window was estimated as the point that performance no longer improved, and was found to be 650ms on average. In Experiment 2, the duration of the visual stimuli was kept short (100ms or <30ms) while the number of photons was varied to explore the efficiency of summation over the integration window compared to Experiment 1. There was some indication that temporal summation remains efficient over the integration window, although there is variation between individuals. The relatively long integration window measured in this study may be relevant to studies of the absolute visual threshold, i.e., tests of single-photon vision, where "single" photons should be separated by greater than the integration window to avoid summation. Copyright © 2017 Elsevier Ltd. All rights reserved.
Curreli, Francesca; Robles, Monica A; Friedman-Kien, Alvin E; Flore, Ornella
2003-02-01
Kaposi's sarcoma-associated herpesvirus is a novel herpesvirus linked to AIDS-related neoplasms. Currently it is difficult to evaluate the number of virions in viral preparation or in samples obtained from patients with Kaposi's sarcoma (KS), since no protocol for determining the plaque forming units of KSHV exists. We constructed a fragment of a different size than the target viral DNA to carry out a competitive-quantitative PCR. Both fragment and viral DNA were added to a single PCR reaction to compete for the same set of primers. By knowing the amount of the competitor added to the reaction, we could determine the number of viral DNA molecules. We used this assay successfully to detect and quantify KSHV genomes from KS skin biopsies and pleural effusion lymphoma, and from different viral preparations. To date, this is the most convenient and economic method that allows an accurate and fast viral detection/quantitation with a single PCR.
Data Randomization and Cluster-Based Partitioning for Botnet Intrusion Detection.
Al-Jarrah, Omar Y; Alhussein, Omar; Yoo, Paul D; Muhaidat, Sami; Taha, Kamal; Kim, Kwangjo
2016-08-01
Botnets, which consist of remotely controlled compromised machines called bots, provide a distributed platform for several threats against cyber world entities and enterprises. Intrusion detection system (IDS) provides an efficient countermeasure against botnets. It continually monitors and analyzes network traffic for potential vulnerabilities and possible existence of active attacks. A payload-inspection-based IDS (PI-IDS) identifies active intrusion attempts by inspecting transmission control protocol and user datagram protocol packet's payload and comparing it with previously seen attacks signatures. However, the PI-IDS abilities to detect intrusions might be incapacitated by packet encryption. Traffic-based IDS (T-IDS) alleviates the shortcomings of PI-IDS, as it does not inspect packet payload; however, it analyzes packet header to identify intrusions. As the network's traffic grows rapidly, not only the detection-rate is critical, but also the efficiency and the scalability of IDS become more significant. In this paper, we propose a state-of-the-art T-IDS built on a novel randomized data partitioned learning model (RDPLM), relying on a compact network feature set and feature selection techniques, simplified subspacing and a multiple randomized meta-learning technique. The proposed model has achieved 99.984% accuracy and 21.38 s training time on a well-known benchmark botnet dataset. Experiment results demonstrate that the proposed methodology outperforms other well-known machine-learning models used in the same detection task, namely, sequential minimal optimization, deep neural network, C4.5, reduced error pruning tree, and randomTree.
Detection of non-classical space-time correlations with a novel type of single-photon camera.
Just, Felix; Filipenko, Mykhaylo; Cavanna, Andrea; Michel, Thilo; Gleixner, Thomas; Taheri, Michael; Vallerga, John; Campbell, Michael; Tick, Timo; Anton, Gisela; Chekhova, Maria V; Leuchs, Gerd
2014-07-14
During the last decades, multi-pixel detectors have been developed capable of registering single photons. The newly developed hybrid photon detector camera has a remarkable property that it has not only spatial but also temporal resolution. In this work, we apply this device to the detection of non-classical light from spontaneous parametric down-conversion and use two-photon correlations for the absolute calibration of its quantum efficiency.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Su, Hui
2001-01-01
Laser-induced fluorescence detection is one of the most sensitive detection techniques and it has found enormous applications in various areas. The purpose of this research was to develop detection approaches based on laser-induced fluorescence detection in two different areas, heterogeneous catalysts screening and single cell study. First, we introduced laser-induced imaging (LIFI) as a high-throughput screening technique for heterogeneous catalysts to explore the use of this high-throughput screening technique in discovery and study of various heterogeneous catalyst systems. This scheme is based on the fact that the creation or the destruction of chemical bonds alters the fluorescence properties of suitablymore » designed molecules. By irradiating the region immediately above the catalytic surface with a laser, the fluorescence intensity of a selected product or reactant can be imaged by a charge-coupled device (CCD) camera to follow the catalytic activity as a function of time and space. By screening the catalytic activity of vanadium pentoxide catalysts in oxidation of naphthalene, we demonstrated LIFI has good detection performance and the spatial and temporal resolution needed for high-throughput screening of heterogeneous catalysts. The sample packing density can reach up to 250 x 250 subunits/cm 2 for 40-μm wells. This experimental set-up also can screen solid catalysts via near infrared thermography detection.« less
Direct observation of single-charge-detection capability of nanowire field-effect transistors.
Salfi, J; Savelyev, I G; Blumin, M; Nair, S V; Ruda, H E
2010-10-01
A single localized charge can quench the luminescence of a semiconductor nanowire, but relatively little is known about the effect of single charges on the conductance of the nanowire. In one-dimensional nanostructures embedded in a material with a low dielectric permittivity, the Coulomb interaction and excitonic binding energy are much larger than the corresponding values when embedded in a material with the same dielectric permittivity. The stronger Coulomb interaction is also predicted to limit the carrier mobility in nanowires. Here, we experimentally isolate and study the effect of individual localized electrons on carrier transport in InAs nanowire field-effect transistors, and extract the equivalent charge sensitivity. In the low carrier density regime, the electrostatic potential produced by one electron can create an insulating weak link in an otherwise conducting nanowire field-effect transistor, modulating its conductance by as much as 4,200% at 31 K. The equivalent charge sensitivity, 4 × 10(-5) e Hz(-1/2) at 25 K and 6 × 10(-5) e Hz(-1/2) at 198 K, is orders of magnitude better than conventional field-effect transistors and nanoelectromechanical systems, and is just a factor of 20-30 away from the record sensitivity for state-of-the-art single-electron transistors operating below 4 K (ref. 8). This work demonstrates the feasibility of nanowire-based single-electron memories and illustrates a physical process of potential relevance for high performance chemical sensors. The charge-state-detection capability we demonstrate also makes the nanowire field-effect transistor a promising host system for impurities (which may be introduced intentionally or unintentionally) with potentially long spin lifetimes, because such transistors offer more sensitive spin-to-charge conversion readout than schemes based on conventional field-effect transistors.
Single-Molecule Plasmon Sensing: Current Status and Future Prospects
2017-01-01
Single-molecule detection has long relied on fluorescent labeling with high quantum-yield fluorophores. Plasmon-enhanced detection circumvents the need for labeling by allowing direct optical detection of weakly emitting and completely nonfluorescent species. This review focuses on recent advances in single molecule detection using plasmonic metal nanostructures as a sensing platform, particularly using a single particle–single molecule approach. In the past decade two mechanisms for plasmon-enhanced single-molecule detection have been demonstrated: (1) by plasmonically enhancing the emission of weakly fluorescent biomolecules, or (2) by monitoring shifts of the plasmon resonance induced by single-molecule interactions. We begin with a motivation regarding the importance of single molecule detection, and advantages plasmonic detection offers. We describe both detection mechanisms and discuss challenges and potential solutions. We finalize by highlighting the exciting possibilities in analytical chemistry and medical diagnostics. PMID:28762723
Raghuram, Jayaram; Miller, David J; Kesidis, George
2014-07-01
We propose a method for detecting anomalous domain names, with focus on algorithmically generated domain names which are frequently associated with malicious activities such as fast flux service networks, particularly for bot networks (or botnets), malware, and phishing. Our method is based on learning a (null hypothesis) probability model based on a large set of domain names that have been white listed by some reliable authority. Since these names are mostly assigned by humans, they are pronounceable, and tend to have a distribution of characters, words, word lengths, and number of words that are typical of some language (mostly English), and often consist of words drawn from a known lexicon. On the other hand, in the present day scenario, algorithmically generated domain names typically have distributions that are quite different from that of human-created domain names. We propose a fully generative model for the probability distribution of benign (white listed) domain names which can be used in an anomaly detection setting for identifying putative algorithmically generated domain names. Unlike other methods, our approach can make detections without considering any additional (latency producing) information sources, often used to detect fast flux activity. Experiments on a publicly available, large data set of domain names associated with fast flux service networks show encouraging results, relative to several baseline methods, with higher detection rates and low false positive rates.
Raghuram, Jayaram; Miller, David J.; Kesidis, George
2014-01-01
We propose a method for detecting anomalous domain names, with focus on algorithmically generated domain names which are frequently associated with malicious activities such as fast flux service networks, particularly for bot networks (or botnets), malware, and phishing. Our method is based on learning a (null hypothesis) probability model based on a large set of domain names that have been white listed by some reliable authority. Since these names are mostly assigned by humans, they are pronounceable, and tend to have a distribution of characters, words, word lengths, and number of words that are typical of some language (mostly English), and often consist of words drawn from a known lexicon. On the other hand, in the present day scenario, algorithmically generated domain names typically have distributions that are quite different from that of human-created domain names. We propose a fully generative model for the probability distribution of benign (white listed) domain names which can be used in an anomaly detection setting for identifying putative algorithmically generated domain names. Unlike other methods, our approach can make detections without considering any additional (latency producing) information sources, often used to detect fast flux activity. Experiments on a publicly available, large data set of domain names associated with fast flux service networks show encouraging results, relative to several baseline methods, with higher detection rates and low false positive rates. PMID:25685511
Multi-feature classifiers for burst detection in single EEG channels from preterm infants
NASA Astrophysics Data System (ADS)
Navarro, X.; Porée, F.; Kuchenbuch, M.; Chavez, M.; Beuchée, Alain; Carrault, G.
2017-08-01
Objective. The study of electroencephalographic (EEG) bursts in preterm infants provides valuable information about maturation or prognostication after perinatal asphyxia. Over the last two decades, a number of works proposed algorithms to automatically detect EEG bursts in preterm infants, but they were designed for populations under 35 weeks of post menstrual age (PMA). However, as the brain activity evolves rapidly during postnatal life, these solutions might be under-performing with increasing PMA. In this work we focused on preterm infants reaching term ages (PMA ⩾36 weeks) using multi-feature classification on a single EEG channel. Approach. Five EEG burst detectors relying on different machine learning approaches were compared: logistic regression (LR), linear discriminant analysis (LDA), k-nearest neighbors (kNN), support vector machines (SVM) and thresholding (Th). Classifiers were trained by visually labeled EEG recordings from 14 very preterm infants (born after 28 weeks of gestation) with 36-41 weeks PMA. Main results. The most performing classifiers reached about 95% accuracy (kNN, SVM and LR) whereas Th obtained 84%. Compared to human-automatic agreements, LR provided the highest scores (Cohen’s kappa = 0.71) using only three EEG features. Applying this classifier in an unlabeled database of 21 infants ⩾36 weeks PMA, we found that long EEG bursts and short inter-burst periods are characteristic of infants with the highest PMA and weights. Significance. In view of these results, LR-based burst detection could be a suitable tool to study maturation in monitoring or portable devices using a single EEG channel.
Danne, Thomas; Bluhmki, Tobias; Seufert, Jochen; Kaltheuner, Matthias; Rathmann, Wolfgang; Beyersmann, Jan; Bramlage, Peter
2015-10-07
In patients with type-2 diabetes receiving oral antidiabetic drugs (OADs), the addition of insulin is frequently required to achieve sufficient control over blood glucose levels. It is, however, difficult to predict if, when and in which patients insulin therapy will be needed. We aimed to identify patient related variables associated with the addition of basal insulin to oral therapy resulting in a basal supported oral therapy (BOT). DIVE (DIabetes Versorgungs-Evaluation) is a prospective, observational, multi-centre diabetes registry established in Germany in 2011. For the present explorative analysis, 31,008 patients with type-2 diabetes prescribed at least one OAD were included. Patients who had previously received insulin and those over 90 years old were excluded. The event of interest was defined as the initiation of BOT during the observational period. Cause-specific Cox proportional hazards models based on a competing risk framework were applied for risk quantification. Multivariable adjusted hazard ratios demonstrated that longer diabetes duration, higher BMI, poorer glycaemic control, documentation of any micro- or macrovascular comorbidity, the presence of concomitant non-antidiabetic pharmacotherapies, and greater numbers of prescribed OADs increased the likelihood of BOT initiation. On the other hand BOT initiation was less likely in patients with older age and female gender. Analysing the likelihood of OAD termination without initiation of BOT provided supportive evidence for the variables predictive of BOT initiation. Analysis of the DIVE registry has resulted in the identification of a number of factors that may be predictive for the initiation of BOT for type-2 diabetes patients initially prescribed one or more OADs. Poor glycaemic control, the presence of vascular comorbidities and concomitant medications, and a greater number of OADs were all detected to increase the risk of a switch to BOT. Female gender and younger age showed protective
Ionization detector, electrode configuration and single polarity charge detection method
He, Z.
1998-07-07
An ionization detector, an electrode configuration and a single polarity charge detection method each utilize a boundary electrode which symmetrically surrounds first and second central interlaced and symmetrical electrodes. All of the electrodes are held at a voltage potential of a first polarity type. The first central electrode is held at a higher potential than the second central or boundary electrodes. By forming the first and second central electrodes in a substantially interlaced and symmetrical pattern and forming the boundary electrode symmetrically about the first and second central electrodes, signals generated by charge carriers are substantially of equal strength with respect to both of the central electrodes. The only significant difference in measured signal strength occurs when the charge carriers move to within close proximity of the first central electrode and are received at the first central electrode. The measured signals are then subtracted and compared to quantitatively measure the magnitude of the charge. 10 figs.
Ionization detector, electrode configuration and single polarity charge detection method
He, Zhong
1998-01-01
An ionization detector, an electrode configuration and a single polarity charge detection method each utilize a boundary electrode which symmetrically surrounds first and second central interlaced and symmetrical electrodes. All of the electrodes are held at a voltage potential of a first polarity type. The first central electrode is held at a higher potential than the second central or boundary electrodes. By forming the first and second central electrodes in a substantially interlaced and symmetrical pattern and forming the boundary electrode symmetrically about the first and second central electrodes, signals generated by charge carriers are substantially of equal strength with respect to both of the central electrodes. The only significant difference in measured signal strength occurs when the charge carriers move to within close proximity of the first central electrode and are received at the first central electrode. The measured signals are then subtracted and compared to quantitatively measure the magnitude of the charge.
A novel single-stranded DNA detection method based on organic semiconductor heterojunction
NASA Astrophysics Data System (ADS)
Gu, Wen; Liu, Hongbo; Zhang, Xia; Zhang, Hao; Chen, Xiong; Wang, Jun
2016-12-01
We demonstrate a novel DNA detection method with low-cost and disposable advantages by utilizing F16CuPc/CuPc planar organic heterojunction device. Single-stranded DNA (ssDNA) molecules have been well immobilized on the surface of CuPc film observed by atomic force microscopy, producing an obvious electrical response of the device. The conductivity of the organic heterojunction film was significantly increased by ssDNA immobilization because ssDNA molecules brought additional positive charges at heterojunction interface. Furthermore, the thickness dependence of CuPc upper layer on the electrical response was studied to optimize the sensitivity. This study will be helpful for the development of organic heterojunction based biosensors.
Detection of four important Eimeria species by multiplex PCR in a single assay.
You, Myung-Jo
2014-06-01
The oocysts of some of the recognized species of chicken coccidiosis are difficult to distinguish morphologically. Diagnostic laboratories are increasingly utilizing DNA-based technologies for the specific identification of Eimeria species. This study reports a multiplex polymerase chain reaction (PCR) assay based on internal transcribed spacer-1 (ITS-1) for the simultaneous diagnosis of the Eimeria tenella, Eimeria acervulina, Eimeria maxima, and Eimeria necatrix species, which infect domestic fowl. Primer pairs specific to each species were designed in order to generate a ladder of amplification products ranging from 20 to 25 bp, and a common optimum annealing temperature for these species was determined to be 52.5 °C. Sensitivity tests were performed for each species, showing a detection threshold of 1-5 pg. All the species were amplified homogeneously, and a homogenous band ladder was observed, indicating that the assay permitted the simultaneous detection of all the species in a single-tube reaction. In the phylogenic study, there was a clear species clustering, which was irrespective of geographical location, for all the ITS-1 sequences used. This multiplex PCR assay represents a rapid and potential cost-effective diagnostic method for the detection of some key Eimeria species that infect domestic fowl. Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.
Material-specific detection and classification of single nanoparticles
Person, Steven; Deutsch, Bradley; Mitra, Anirban; Novotny, Lukas
2010-01-01
Detection and classification of nanoparticles is important for environmental monitoring, contamination mitigation, biological label tracking, and bio-defense. Detection techniques involve a trade-off between sensitivity, discrimination, and speed. This paper presents a material-specific dual-color common-path interferometric detection system. Two wavelengths are simultaneously used to discriminate between 60 nm silver and 80 nm diameter gold particles in solution with a detection time of τ ≈ 1 ms. The detection technique is applicable to situations where both particle size and material are of interest. PMID:21142033
A versatile optical microscope for time-dependent single-molecule and single-particle spectroscopy
NASA Astrophysics Data System (ADS)
Li, Hao; Yang, Haw
2018-03-01
This work reports the design and implementation of a multi-function optical microscope for time-dependent spectroscopy on single molecules and single nanoparticles. It integrates the now-routine single-object measurements into one standalone platform so that no reconfiguration is needed when switching between different types of sample or spectroscopy modes. The illumination modes include evanescent field through total internal reflection, dark-field illumination, and epi-excitation onto a diffraction-limited spot suitable for confocal detection. The detection modes include spectrally resolved line imaging, wide-field imaging with dual-color capability, and two-color single-element photon-counting detection. The switch between different spectroscopy and data acquisition modes is fully automated and executed through computer programming. The capability of this microscope is demonstrated through selected proof-of-principle experiments.
A versatile optical microscope for time-dependent single-molecule and single-particle spectroscopy.
Li, Hao; Yang, Haw
2018-03-28
This work reports the design and implementation of a multi-function optical microscope for time-dependent spectroscopy on single molecules and single nanoparticles. It integrates the now-routine single-object measurements into one standalone platform so that no reconfiguration is needed when switching between different types of sample or spectroscopy modes. The illumination modes include evanescent field through total internal reflection, dark-field illumination, and epi-excitation onto a diffraction-limited spot suitable for confocal detection. The detection modes include spectrally resolved line imaging, wide-field imaging with dual-color capability, and two-color single-element photon-counting detection. The switch between different spectroscopy and data acquisition modes is fully automated and executed through computer programming. The capability of this microscope is demonstrated through selected proof-of-principle experiments.
Single photon detection with self-quenching multiplication
NASA Technical Reports Server (NTRS)
Zheng, Xinyu (Inventor); Cunningham, Thomas J. (Inventor); Pain, Bedabrata (Inventor)
2011-01-01
A photoelectronic device and an avalanche self-quenching process for a photoelectronic device are described. The photoelectronic device comprises a nanoscale semiconductor multiplication region and a nanoscale doped semiconductor quenching structure including a depletion region and an undepletion region. The photoelectronic device can act as a single photon detector or a single carrier multiplier. The avalanche self-quenching process allows electrical field reduction in the multiplication region by movement of the multiplication carriers, thus quenching the avalanche.
NASA Astrophysics Data System (ADS)
Wakui, Kentaro; Hayasaka, Kazuhiro; Ido, Tetsuya
2014-12-01
Vacuum ultraviolet (VUV) radiation around 159 nm is obtained toward direct excitation of a single trapped ion. An efficient fluoride-based VUV output coupler is employed for intracavity high-harmonic generation of a Ti:S oscillator. Using this coupler, where we measured its reflectance to be about 90 %, an average power reaching 6.4 W is coupled out from a modest fundamental power of 650 mW. When a single comb component out of 1.9 10 teeth is resonant to the atomic transition, 100s of fluorescence photons per second will be detectable under a realistic condition.
Detecting trace pesticides in real time using single particle aerosol mass spectrometry.
Barker, Zachary; Venkatchalam, Veena; Martin, Audrey N; Farquar, George R; Frank, Matthias
2010-02-28
Pesticides are toxic substances and may cause unintentional harm if improperly used. The ubiquitous nature of pesticides, with frequent use in agriculture and the household, and the potential for harm that pesticides pose to non-target organisms such as wildlife, humans, and pets, demonstrate the need for rapid and effective detection and identification of these compounds. In this study, single particle aerosol mass spectrometry (SPAMS) was used to rapidly detect compounds from four classes of pesticides commonly used in agricultural and household applications. These include permethrin (pyrethroid class), malathion and dichlorvos (organophosphate class), imidacloprid (chloronicotinyl class), and carbaryl (carbamate class). Analytical standards of each compound were diluted and aerosolized using a nebulizer to create particles for analysis in the SPAMS instrument. The resultant dual-polarity time-of-flight mass spectra were then analyzed to identify the characteristic peaks of the compound in each sample. In addition, samples of commercial products containing pesticides, a commercial insecticide spray, containing permethrin, and a canine flea collar, containing carbaryl, were analyzed in their original form using SPAMS without any significant sample preparation. The characteristic mass spectral peaks of the active pesticides in these samples were identified using the mass spectra obtained earlier from the pesticide analytical standards. By successfully identifying pesticides in analytical standards and in commercial products, it is demonstrated herein that the SPAMS system may be capable of pesticide detection in numerous environmental and agricultural situations. Copyright 2010 Elsevier B.V. All rights reserved.
Haugum, K; Brandal, L T; Løbersli, I; Kapperud, G; Lindstedt, B-A
2011-06-01
To compare 167 Norwegian human and nonhuman Escherichia coli O157:H7/NM (nonmotile) isolates with respect to an A/T single nucleotide polymorphism (SNP) in the tir gene and to detect specific SNPs that differentiate STEC O157 into distinct virulence clades (1-3 and 8). We developed a multiplex PCR followed by single base sequencing for detection of the SNPs, and examined the association among SNP genotype, virulence profile (stx and eae status), multilocus variable number of tandem repeats analysis (MLVA) profile and clinical outcome. We found an over-representation of the T allele among human strains compared to nonhuman strains, including 5/6 haemolytic-uraemic syndrome cases. Fourteen strains belonged to clade 8, followed by two clade 2 strains. No clade 1 nor 3 isolates were observed. stx1 in combination with either stx2(EDL933) or stx2c were frequently observed among human strains, whereas stx2c was dominating in nonhuman strains. MLVA indicated that only single cases or small outbreaks with E. coli O157 have been observed in Norway through the years 1993-2008. We observed that the tir-255 A/T SNP and the stx status were different between human and nonhuman O157 strains. No major outbreaks were observed, and only a few strains were differentiated into the virulence clades 2 and 8. The detection of virulence clade-specific SNPs enables the rapid designation of virulent E. coli O157 strains, especially in outbreak situations. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.
Ballari, Rajashekhar V; Martin, Asha; Gowda, Lalitha R
2013-01-01
Brinjal is an important vegetable crop. Major crop loss of brinjal is due to insect attack. Insect-resistant EE-1 brinjal has been developed and is awaiting approval for commercial release. Consumer health concerns and implementation of international labelling legislation demand reliable analytical detection methods for genetically modified (GM) varieties. End-point and real-time polymerase chain reaction (PCR) methods were used to detect EE-1 brinjal. In end-point PCR, primer pairs specific to 35S CaMV promoter, NOS terminator and nptII gene common to other GM crops were used. Based on the revealed 3' transgene integration sequence, primers specific for the event EE-1 brinjal were designed. These primers were used for end-point single, multiplex and SYBR-based real-time PCR. End-point single PCR showed that the designed primers were highly specific to event EE-1 with a sensitivity of 20 pg of genomic DNA, corresponding to 20 copies of haploid EE-1 brinjal genomic DNA. The limits of detection and quantification for SYBR-based real-time PCR assay were 10 and 100 copies respectively. The prior development of detection methods for this important vegetable crop will facilitate compliance with any forthcoming labelling regulations. Copyright © 2012 Society of Chemical Industry.
NASA Astrophysics Data System (ADS)
Choi, Sam B.; Lombard-Banek, Camille; Muñoz-LLancao, Pablo; Manzini, M. Chiara; Nemes, Peter
2018-05-01
The ability to detect peptides and proteins in single cells is vital for understanding cell heterogeneity in the nervous system. Capillary electrophoresis (CE) nanoelectrospray ionization (nanoESI) provides high-resolution mass spectrometry (HRMS) with trace-level sensitivity, but compressed separation during CE challenges protein identification by tandem HRMS with limited MS/MS duty cycle. Here, we supplemented ultrasensitive CE-nanoESI-HRMS with reversed-phase (RP) fractionation to enhance identifications from protein digest amounts that approximate to a few mammalian neurons. An 1 to 20 μg neuronal protein digest was fractionated on a RP column (ZipTip), and 1 ng to 500 pg of peptides were analyzed by a custom-built CE-HRMS system. Compared with the control (no fractionation), RP fractionation improved CE separation (theoretical plates 274,000 versus 412,000 maximum, resp.), which enhanced detection sensitivity (2.5-fold higher signal-to-noise ratio), minimized co-isolation spectral interferences during MS/MS, and increased the temporal rate of peptide identification by up to 57%. From 1 ng of protein digest (<5 neurons), CE with RP fractionation identified 737 protein groups (1,753 peptides), or 480 protein groups ( 1,650 peptides) on average per analysis. The approach was scalable to 500 pg of protein digest ( a single neuron), identifying 225 protein groups (623 peptides) in technical triplicates, or 141 protein groups on average per analysis. Among identified proteins, 101 proteins were products of genes that are known to be transcriptionally active in single neurons during early development of the brain, including those involved in synaptic transmission and plasticity and cytoskeletal organization. [Figure not available: see fulltext.
NASA Astrophysics Data System (ADS)
Choi, Sam B.; Lombard-Banek, Camille; Muñoz-LLancao, Pablo; Manzini, M. Chiara; Nemes, Peter
2017-11-01
The ability to detect peptides and proteins in single cells is vital for understanding cell heterogeneity in the nervous system. Capillary electrophoresis (CE) nanoelectrospray ionization (nanoESI) provides high-resolution mass spectrometry (HRMS) with trace-level sensitivity, but compressed separation during CE challenges protein identification by tandem HRMS with limited MS/MS duty cycle. Here, we supplemented ultrasensitive CE-nanoESI-HRMS with reversed-phase (RP) fractionation to enhance identifications from protein digest amounts that approximate to a few mammalian neurons. An 1 to 20 μg neuronal protein digest was fractionated on a RP column (ZipTip), and 1 ng to 500 pg of peptides were analyzed by a custom-built CE-HRMS system. Compared with the control (no fractionation), RP fractionation improved CE separation (theoretical plates 274,000 versus 412,000 maximum, resp.), which enhanced detection sensitivity (2.5-fold higher signal-to-noise ratio), minimized co-isolation spectral interferences during MS/MS, and increased the temporal rate of peptide identification by up to 57%. From 1 ng of protein digest (<5 neurons), CE with RP fractionation identified 737 protein groups (1,753 peptides), or 480 protein groups ( 1,650 peptides) on average per analysis. The approach was scalable to 500 pg of protein digest ( a single neuron), identifying 225 protein groups (623 peptides) in technical triplicates, or 141 protein groups on average per analysis. Among identified proteins, 101 proteins were products of genes that are known to be transcriptionally active in single neurons during early development of the brain, including those involved in synaptic transmission and plasticity and cytoskeletal organization. [Figure not available: see fulltext.
Ping Kong; Patricia A. Richardson; Chuanxue Hong; Thomas L. Kubisiak
2006-01-01
At the first Sudden Oak Death Science Symposium, we reported on the use of a single strand conformation polymorphism (SSCP) analysis for rapid identification of Phytophthora ramorum in culture. We have since assessed and improved the fingerprinting technique for detecting this pathogen directly from plant tissues. The improved SSCP protocol uses a...
Yip, Mary; Saripan, M Iqbal; Wells, Kevin; Bradley, David A
2015-01-01
Detection of buried improvised explosive devices (IEDs) is a delicate task, leading to a need to develop sensitive stand-off detection technology. The shape, composition and size of the IEDs can be expected to be revised over time in an effort to overcome increasingly sophisticated detection methods. As an example, for the most part, landmines are found through metal detection which has led to increasing use of non-ferrous materials such as wood or plastic containers for chemical based explosives being developed. Monte Carlo simulations have been undertaken considering three different commercially available detector materials (hyperpure-Ge (HPGe), lanthanum(III) bromide (LaBr) and thallium activated sodium iodide (NaI(Tl)), applied at a stand-off distance of 50 cm from the surface and burial depths of 0, 5 and 10 cm, with sand as the obfuscating medium. Target materials representing medium density wood and mild steel have been considered. Each detector has been modelled as a 10 cm thick cylinder with a 20 cm diameter. It appears that HPGe represents the most promising detector for this application. Although it was not the highest density material studied, its excellent energy resolving capability leads to the highest quality spectra from which detection decisions can be inferred. The simulation work undertaken here suggests that a vehicle-born threat detection system could be envisaged using a single betatron and a series of detectors operating in parallel observing the space directly in front of the vehicle path. Furthermore, results show that non-ferrous materials such as wood can be effectively discerned in such remote-operated detection system, with the potential to apply a signature analysis template matching technique for real-time analysis of such data.
Hardware solution for continuous time-resolved burst detection of single molecules in flow
NASA Astrophysics Data System (ADS)
Wahl, Michael; Erdmann, Rainer; Lauritsen, Kristian; Rahn, Hans-Juergen
1998-04-01
Time Correlated Single Photon Counting (TCSPC) is a valuable tool for Single Molecule Detection (SMD). However, existing TCSPC systems did not support continuous data collection and processing as is desirable for applications such as SMD for e.g. DNA-sequencing in a liquid flow. First attempts at using existing instrumentation in this kind of operation mode required additional routing hardware to switch between several memory banks and were not truly continuous. We have designed a hard- and software system to perform continuous real-time TCSPC based upon a modern solid state Time to Digital Converter (TDC). Short dead times of the fully digital TDC design combined with fast Field Programmable Gay Array logic permit a continuous data throughput as high as 3 Mcounts/sec. The histogramming time may be set as short as 100 microsecond(s) . Every histogram or every single fluorescence photon can be real-time tagged at 200 ns resolution in addition to recording its arrival time relative to the excitation pulse. Continuous switching between memory banks permits concurrent histogramming and data read-out. The instrument provides a time resolution of 60 ps and up to 4096 histogram channels. The overall instrument response function in combination with a low cost picosecond diode laser and an inexpensive photomultiplier tube was found to be 180 ps and well sufficient to measure sub-nanosecond fluorescence lifetimes.
Shen, Rui; Liu, Peipei; Zhang, Yiqiu; Yu, Zhao; Chen, Xuyue; Zhou, Lu; Nie, Baoqing; Żaczek, Anna; Chen, Jian; Liu, Jian
2018-04-03
As an important signaling molecule, hydrogen peroxide (H 2 O 2 ) secreted externally by the cells influences cell migration, immunity generation, and cellular communications. Herein, we have developed a microfluidic approach with droplets in combination with Au nanoclusters for the sensitive detection of H 2 O 2 secreted by a single cell. Isolated in the ultrasmall volume (4.2 nL) of a microdroplet, single-cell secreted H 2 O 2 can initiate dramatic fluorescence changes of horseradish peroxidase-Au nanoclusters. We have demonstrated an ultrahigh sensitivity (200-400 attomole H 2 O 2 directly measured from a single cell) with good specificity. It offers a useful research tool to study the cell-to-cell differences of H 2 O 2 secretion at the single-cell level.
smiFISH and FISH-quant - a flexible single RNA detection approach with super-resolution capability.
Tsanov, Nikolay; Samacoits, Aubin; Chouaib, Racha; Traboulsi, Abdel-Meneem; Gostan, Thierry; Weber, Christian; Zimmer, Christophe; Zibara, Kazem; Walter, Thomas; Peter, Marion; Bertrand, Edouard; Mueller, Florian
2016-12-15
Single molecule FISH (smFISH) allows studying transcription and RNA localization by imaging individual mRNAs in single cells. We present smiFISH (single molecule inexpensive FISH), an easy to use and flexible RNA visualization and quantification approach that uses unlabelled primary probes and a fluorescently labelled secondary detector oligonucleotide. The gene-specific probes are unlabelled and can therefore be synthesized at low cost, thus allowing to use more probes per mRNA resulting in a substantial increase in detection efficiency. smiFISH is also flexible since differently labelled secondary detector probes can be used with the same primary probes. We demonstrate that this flexibility allows multicolor labelling without the need to synthesize new probe sets. We further demonstrate that the use of a specific acrydite detector oligonucleotide allows smiFISH to be combined with expansion microscopy, enabling the resolution of transcripts in 3D below the diffraction limit on a standard microscope. Lastly, we provide improved, fully automated software tools from probe-design to quantitative analysis of smFISH images. In short, we provide a complete workflow to obtain automatically counts of individual RNA molecules in single cells. © The Author(s) 2016. Published by Oxford University Press on behalf of Nucleic Acids Research.
Chen, Zhongxue; Ng, Hon Keung Tony; Li, Jing; Liu, Qingzhong; Huang, Hanwen
2017-04-01
In the past decade, hundreds of genome-wide association studies have been conducted to detect the significant single-nucleotide polymorphisms that are associated with certain diseases. However, most of the data from the X chromosome were not analyzed and only a few significant associated single-nucleotide polymorphisms from the X chromosome have been identified from genome-wide association studies. This is mainly due to the lack of powerful statistical tests. In this paper, we propose a novel statistical approach that combines the information of single-nucleotide polymorphisms on the X chromosome from both males and females in an efficient way. The proposed approach avoids the need of making strong assumptions about the underlying genetic models. Our proposed statistical test is a robust method that only makes the assumption that the risk allele is the same for both females and males if the single-nucleotide polymorphism is associated with the disease for both genders. Through simulation study and a real data application, we show that the proposed procedure is robust and have excellent performance compared to existing methods. We expect that many more associated single-nucleotide polymorphisms on the X chromosome will be identified if the proposed approach is applied to current available genome-wide association studies data.
A CMOS enhanced solid-state nanopore based single molecule detection platform.
Chen, Chinhsuan; Yemenicioglu, Sukru; Uddin, Ashfaque; Corgliano, Ellie; Theogarajan, Luke
2013-01-01
Solid-state nanopores have emerged as a single molecule label-free electronic detection platform. Existing transimpedance stages used to measure ionic current nanopores suffer from dynamic range limitations resulting from steady-state baseline currents. We propose a digitally-assisted baseline cancellation CMOS platform that circumvents this issue. Since baseline cancellation is a form of auto-zeroing, the 1/f noise of the system is also reduced. Our proposed design can tolerate a steady state baseline current of 10µA and has a usable bandwidth of 750kHz. Quantitative DNA translocation experiments on 5kbp DNA was performed using a 5nm silicon nitride pore using both the CMOS platform and a commercial system. Comparison of event-count histograms show that the CMOS platform clearly outperforms the commercial system, allowing for unambiguous interpretation of the data.
Lin, Zhimin; Zeng, Ying; Tong, Li; Zhang, Hangming; Zhang, Chi
2017-01-01
The application of electroencephalogram (EEG) generated by human viewing images is a new thrust in image retrieval technology. A P300 component in the EEG is induced when the subjects see their point of interest in a target image under the rapid serial visual presentation (RSVP) experimental paradigm. We detected the single-trial P300 component to determine whether a subject was interested in an image. In practice, the latency and amplitude of the P300 component may vary in relation to different experimental parameters, such as target probability and stimulus semantics. Thus, we proposed a novel method, Target Recognition using Image Complexity Priori (TRICP) algorithm, in which the image information is introduced in the calculation of the interest score in the RSVP paradigm. The method combines information from the image and EEG to enhance the accuracy of single-trial P300 detection on the basis of traditional single-trial P300 detection algorithm. We defined an image complexity parameter based on the features of the different layers of a convolution neural network (CNN). We used the TRICP algorithm to compute for the complexity of an image to quantify the effect of different complexity images on the P300 components and training specialty classifier according to the image complexity. We compared TRICP with the HDCA algorithm. Results show that TRICP is significantly higher than the HDCA algorithm (Wilcoxon Sign Rank Test, p<0.05). Thus, the proposed method can be used in other and visual task-related single-trial event-related potential detection. PMID:29283998
Charles, Paul T; Davis, Jasmine; Adams, André A; Anderson, George P; Liu, Jinny L; Deschamps, Jeffrey R; Kusterbeck, Anne W
2015-11-01
The development of explosives detection technologies has increased significantly over the years as environmental and national security agencies implement tighter pollution control measures and methods for improving homeland security. 2, 4, 6-Trinitrotoluene (TNT), known primarily as a component in munitions, has been targeted for both its toxicity and carcinogenic properties that if present at high concentrations can be a detriment to both humans, marine and plant ecosystems. Enabling end users with environmental detection and monitoring systems capable of providing real-time, qualitative and quantitative chemical analysis of these toxic compounds would be extremely beneficial. Reported herein is the development of a multi-channeled microfluidic device immobilized with single chain fragment variable (scFv) recombinant proteins specific for the explosive, TNT. Fluorescence displacement immunoassays performed under constant flow demonstrated trace level sensitivity and specificity for TNT. The utility of three multi-channeled devices immobilized with either (1) scFv recombinant protein, (2) biotinylated-scFv (bt-scFv) and (3) monoclonal anti-TNT (whole IgG molecule) were investigated and compared. Fluorescence dose response curves, crossreactivity measurements and limits of detection (LOD) for TNT were determined. Fluorescence displacement immunoassays for TNT in natural seawater demonstrated detection limits at sub-parts-per-billion levels (0.5 ppb) utilizing the microfluidic device with immobilized bt-scFv. Published by Elsevier B.V.
Single-silicon CCD-CMOS platform for multi-spectral detection from terahertz to x-rays.
Shalaby, Mostafa; Vicario, Carlo; Hauri, Christoph P
2017-11-15
Charge-coupled devices (CCDs) are a well-established imaging technology in the visible and x-ray frequency ranges. However, the small quantum photon energies of terahertz radiation have hindered the use of this mature semiconductor technological platform in this frequency range, leaving terahertz imaging totally dependent on low-resolution bolometer technologies. Recently, it has been shown that silicon CCDs can detect terahertz photons at a high field, but the detection sensitivity is limited. Here we show that silicon, complementary metal-oxide-semiconductor (CMOS) technology offers enhanced detection sensitivity of almost two orders of magnitude, compared to CCDs. Our findings allow us to extend the low-frequency terahertz cutoff to less than 2 THz, nearly closing the technological gap with electronic imagers operating up to 1 THz. Furthermore, with the silicon CCD/CMOS technology being sensitive to mid-infrared (mid-IR) and the x-ray ranges, we introduce silicon as a single detector platform from 1 EHz to 2 THz. This overcomes the present challenge in spatially overlapping a terahertz/mid-IR pump and x-ray probe radiation at facilities such as free electron lasers, synchrotron, and laser-based x-ray sources.
Comment on ’Single Pentacene Molecules Detected by Fluorescence Excitation in a P-Terphenyl Crystal’
1990-12-10
8217 NO 11 TITLE (include Security Classification) Comment on "Single Pentacene Molecules Detected by Fluorescence Excitation in a p-Terphenyl Crystal" 12...8217 {Continue on reverse it necessary and identify by block numboer) Using h--,Ihly efficient Fluorescence excitation spectroscov of individual pentacene ...molecular impurities in p-terphenvl crystals, we have observed that some pentacene defects exhibit spcntaneous spectral jumps in their resonance frequency at
Vásquez, Gersson; Rey, Alba; Rivera, Camilo; Iregui, Carlos; Orozco, Jahir
2017-01-15
Pathogenic bacteria are responsible for several diseases in humans and in a variety of hosts. Detection of pathogenic bacteria is imperative to avoid and/or fight their potential harmful effects. This work reports on the first amperometric biosensor for the rapid detection of Streptococcus agalactiae (S. agalactiae). The biosensor relies on a single biotinylated antibody that immobilizes the bacteria on a screen-printed carbon electrode while is further linked to a streptavidin-conjugated HRP reporter. The biotinylated antibody provides selectivity to the biosensor whereas serves as an anchoring point to the reporter for further amplification of the electrochemical signal. The resultant immunosensor is simple, responds rapidly, and allows for the selective and highly sensitive quantification of S. agalactiae cells in a concentration range of 10 1 -10 7 CFUml -1 , with a detection limit of 10CFUml -1 . The approach not only enables a rapid detection and quantification of S. agalactiae in environmental samples but also opens up new opportunities for the simple fabrication of electrochemical immunosensors for different target pathogens. Copyright © 2016 Elsevier B.V. All rights reserved.
Wang, Junsheng; Sun, Jinyang; Song, Yongxin; Xu, Yongyi; Pan, Xinxiang; Sun, Yeqing; Li, Dongqing
2013-01-01
Detection of living microalgae cells is very important for ballast water treatment and analysis. Chlorophyll fluorescence is an indicator of photosynthetic activity and hence the living status of plant cells. In this paper, we developed a novel microfluidic biosensor system that can quickly and accurately detect the viability of single microalgae cells based on chlorophyll fluorescence. The system is composed of a laser diode as an excitation light source, a photodiode detector, a signal analysis circuit, and a microfluidic chip as a microalgae cell transportation platform. To demonstrate the utility of this system, six different living and dead algae samples (Karenia mikimotoi Hansen, Chlorella vulgaris, Nitzschia closterium, Platymonas subcordiformis, Pyramidomonas delicatula and Dunaliella salina) were tested. The developed biosensor can distinguish clearly between the living microalgae cells and the dead microalgae cells. The smallest microalgae cells that can be detected by using this biosensor are 3 μm ones. Even smaller microalgae cells could be detected by increasing the excitation light power. The developed microfluidic biosensor has great potential for in situ ballast water analysis. PMID:24287532
Direct detection of a single evoked action potential with MRS in Lumbricus terrestris.
Poplawsky, Alexander J; Dingledine, Raymond; Hu, Xiaoping P
2012-01-01
Functional MRI (fMRI) measures neural activity indirectly by detecting the signal change associated with the hemodynamic response following brain activation. In order to alleviate the temporal and spatial specificity problems associated with fMRI, a number of attempts have been made to detect neural magnetic fields (NMFs) with MRI directly, but have thus far provided conflicting results. In this study, we used MR to detect axonal NMFs in the median giant fiber of the earthworm, Lumbricus terrestris, by examining the free induction decay (FID) with a sampling interval of 0.32 ms. The earthworm nerve cords were isolated from the vasculature and stimulated at the threshold of action potential generation. FIDs were acquired shortly after the stimulation, and simultaneous field potential recordings identified the presence or absence of single evoked action potentials. FIDs acquired when the stimulus did not evoke an action potential were summed as background. The phase of the background-subtracted FID exhibited a systematic change, with a peak phase difference of (-1.2 ± 0.3) × 10(-5) radians occurring at a time corresponding to the timing of the action potential. In addition, we calculated the possible changes in the FID magnitude and phase caused by a simulated action potential using a volume conductor model. The measured phase difference matched the theoretical prediction well in both amplitude and temporal characteristics. This study provides the first evidence for the direct detection of a magnetic field from an evoked action potential using MR. Copyright © 2011 John Wiley & Sons, Ltd.
King, J.R.; Faugeras, F.; Gramfort, A.; Schurger, A.; El Karoui, I.; Sitt, J.D.; Rohaut, B.; Wacongne, C.; Labyt, E.; Bekinschtein, T.; Cohen, L.; Naccache, L.; Dehaene, S.
2017-01-01
Detecting residual consciousness in unresponsive patients is a major clinical concern and a challenge for theoretical neuroscience. To tackle this issue, we recently designed a paradigm that dissociates two electro-encephalographic (EEG) responses to auditory novelty. Whereas a local change in pitch automatically elicits a mismatch negativity (MMN), a change in global sound sequence leads to a late P300b response. The latter component is thought to be present only when subjects consciously perceive the global novelty. Unfortunately, it can be difficult to detect because individual variability is high, especially in clinical recordings. Here, we show that multivariate pattern classifiers can extract subject-specific EEG patterns and predict single-trial local or global novelty responses. We first validate our method with 38 high-density EEG, MEG and intracranial EEG recordings. We empirically demonstrate that our approach circumvents the issues associated with multiple comparisons and individual variability while improving the statistics. Moreover, we confirm in control subjects that local responses are robust to distraction whereas global responses depend on attention. We then investigate 104 vegetative state (VS), minimally conscious state (MCS) and conscious state (CS) patients recorded with high-density EEG. For the local response, the proportion of significant decoding scores (M = 60%) does not vary with the state of consciousness. By contrast, for the global response, only 14% of the VS patients' EEG recordings presented a significant effect, compared to 31% in MCS patients' and 52% in CS patients'. In conclusion, single-trial multivariate decoding of novelty responses provides valuable information in non-communicating patients and paves the way towards real-time monitoring of the state of consciousness. PMID:23859924
Cao, Yingchun; Sanchez, Nancy P; Jiang, Wenzhe; Griffin, Robert J; Xie, Feng; Hughes, Lawrence C; Zah, Chung-en; Tittel, Frank K
2015-02-09
A continuous wave (CW) quantum cascade laser (QCL) based absorption sensor system was demonstrated and developed for simultaneous detection of atmospheric nitrous oxide (N(2)O), methane (CH(4)), and water vapor (H(2)O). A 7.73-µm CW QCL with its wavelength scanned over a spectral range of 1296.9-1297.6 cm(-1) was used to simultaneously target three neighboring strong absorption lines, N(2)O at 1297.05 cm(-1), CH(4) at 1297.486 cm(-1), and H(2)O at 1297.184 cm(-1). An astigmatic multipass Herriott cell with a 76-m path length was utilized for laser based gas absorption spectroscopy at an optimum pressure of 100 Torr. Wavelength modulation and second harmonic detection was employed for data processing. Minimum detection limits (MDLs) of 1.7 ppb for N(2)O, 8.5 ppb for CH(4), and 11 ppm for H(2)O were achieved with a 2-s integration time for individual gas detection. This single QCL based multi-gas detection system possesses applications in environmental monitoring and breath analysis.
Kinde, Hailu; Goodluck, Helen A; Pitesky, Maurice; Friend, Tom D; Campbell, James A; Hill, Ashley E
2015-12-01
Single swabs (cultured individually) are currently used in the Food and Drug Administration (FDA) official method for sampling the environment of commercial laying hens for the detection of Salmonella enterica ssp. serovar Enteritidis (Salmonella Enteritidis). The FDA has also granted provisional acceptance of the National Poultry Improvement Plan's (NPIP) Salmonella isolation and identification methodology for samples taken from table-egg layer flock environments. The NPIP method, as with the FDA method, requires single-swab culturing for the environmental sampling of laying houses for Salmonella Enteritidis. The FDA culture protocol requires a multistep culture enrichment broth, and it is more labor intensive than the NPIP culture protocol, which requires a single enrichment broth. The main objective of this study was to compare the FDA single-swab culturing protocol with that of the NPIP culturing protocol but using a four-swab pool scheme. Single and multi-laboratory testing of replicate manure drag swab sets (n = 525 and 672, respectively) collected from a Salmonella Enteritidis-free commercial poultry flock was performed by artificially contaminating swabs with either Salmonella Enteritidis phage type 4, 8, or 13a at one of two inoculation levels: low, x¯ = 2.5 CFU (range 2.5-2.7), or medium, x¯ = 10.0 CFU (range 7.5-12). For each replicate, a single swab (inoculated), sets of two swabs (one inoculated and one uninoculated), and sets of four swabs (one inoculated and three uninoculated), testing was conducted using the FDA or NPIP culture method. For swabs inoculated with phage type 8, the NPIP method was more efficient (P < 0.05) for all swab sets at both inoculation levels than the reference method. The single swabs in the NPIP method were significantly (P < 0.05) better than four-pool swabs in detecting Salmonella Enteritidis at the lower inoculation level. In the collaborative study (n = 13 labs) using Salmonella Enteritidis phage
NASA Astrophysics Data System (ADS)
Homberg, C.; Bergerat, F.; Angelier, J.; Garcia, S.
2010-02-01
Transform motion along oceanic transforms generally occurs along narrow faults zones. Another class of oceanic transforms exists where the plate boundary is quite large (˜100 km) and includes several subparallel faults. Using a 2-D numerical modeling, we simulate the slip distribution and the crustal stress field geometry within such broad oceanic transforms (BOTs). We examine the possible configurations and evolution of such BOTs, where the plate boundary includes one, two, or three faults. Our experiments show that at any time during the development of the plate boundary, the plate motion is not distributed along each of the plate boundary faults but mainly occurs along a single master fault. The finite width of a BOT results from slip transfer through time with locking of early faults, not from a permanent distribution of deformation over a wide area. Because of fault interaction, the stress field geometry within the BOTs is more complex than that along classical oceanic transforms and includes stress deflections close to but also away from the major faults. Application of this modeling to the 100 km wide Tjörnes Fracture Zone (TFZ) in North Iceland, a major BOT of the Mid-Atlantic Ridge that includes three main faults, suggests that the Dalvik Fault and the Husavik-Flatey Fault developed first, the Grismsey Fault being the latest active structure. Since initiation of the TFZ, the Husavik-Flatey Fault accommodated most of the plate motion and probably persists until now as the main plate structure.
Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals.
Liu, Feng; Liu, Nannan
2016-01-18
The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house single or multiple neurons inside, are considered to make the major contribution in responding to the chemical stimuli. By directly recording action potential in the olfactory sensillum after exposure to stimuli, single sensillum recording (SSR) technique provides a powerful approach for investigating the neural responses of insects to chemical stimuli. For the bed bug, which is a notorious human parasite, multiple types of olfactory sensillum have been characterized. In this study, we demonstrated neural responses of bed bug olfactory sensilla to two chemical stimuli and the dose-dependent responses to one of them using the SSR method. This approach enables researchers to conduct early screening for individual chemical stimuli on the bed bug olfactory sensilla, which would provide valuable information for the development of new bed bug attractants or repellents and benefits the bed bug control efforts.
Using Single Sensillum Recording to Detect Olfactory Neuron Responses of Bed Bugs to Semiochemicals
Liu, Feng; Liu, Nannan
2016-01-01
The insect olfactory system plays an important role in detecting semiochemicals in the environment. In particular, the antennal sensilla which house single or multiple neurons inside, are considered to make the major contribution in responding to the chemical stimuli. By directly recording action potential in the olfactory sensillum after exposure to stimuli, single sensillum recording (SSR) technique provides a powerful approach for investigating the neural responses of insects to chemical stimuli. For the bed bug, which is a notorious human parasite, multiple types of olfactory sensillum have been characterized. In this study, we demonstrated neural responses of bed bug olfactory sensilla to two chemical stimuli and the dose-dependent responses to one of them using the SSR method. This approach enables researchers to conduct early screening for individual chemical stimuli on the bed bug olfactory sensilla, which would provide valuable information for the development of new bed bug attractants or repellents and benefits the bed bug control efforts. PMID:26862929
Faklaris, Orestis; Garrot, Damien; Joshi, Vandana; Druon, Frédéric; Boudou, Jean-Paul; Sauvage, Thierry; Georges, Patrick; Curmi, Patrick A; Treussart, François
2008-12-01
Diamond nanoparticles are promising photoluminescent probes for tracking intracellular processes, due to embedded, perfectly photostable color centers. In this work, the spontaneous internalization of such nanoparticles (diameter 25 nm) in HeLa cancer cells is investigated by confocal microscopy and time-resolved techniques. Nanoparticles are observed inside the cell cytoplasm at the single-particle and single-color-center level, assessed by time-correlation intensity measurements. Improvement of the nanoparticle signal-to-noise ratio inside the cell is achieved using a pulsed-excitation laser and time-resolved detection taking advantage of the long radiative lifetime of the color-center excited state as compared to cell autofluorescence. The internalization pathways are also investigated, with endosomal marking and colocalization analyses. The low colocalization ratio observed proves that nanodiamonds are not trapped in endosomes, a promising result in prospect of drug delivery by these nanoparticles. Low cytotoxicity of these nanoparticles in this cell line is also shown.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Zotova, A. N., E-mail: zotova@imp.sci-nnov.ru
2016-05-15
The contribution of bends and constrictions by a superconducting film to the detection by a single photon superconducting detector is investigated. It has been shown that, for currents smaller than the minimal detection current of a straight film, the detection efficiency of a film with a constriction attains saturation upon an increase in the current, which coincides qualitatively with the behavior of this dependence observed in the experiment. It has also been found that the effect of bends in the film and the external magnetic field on the detection efficiency for low-energy photons is essential, while for high-energy photons nomore » such influence is observed.« less
NASA Astrophysics Data System (ADS)
Lin, Pei; Liu, Xi; Yan, Xiaoqin; Kang, Zhuo; Lei, Yang; Zhao, Yanguang
2012-08-01
Qualitative and quantitative detection of biological and chemical species is crucial in many areas, ranging from clinical diagnosis to homeland security. Due to the advantages of ultrahigh sensitivity, label-free, fast readout and easy fabrication over the traditional detection systems, semiconductor nanowire based electronic devices have emerged as a potential platform. In this paper, we fabricated a single ZnO nanowire-based bioFET sensor for the detection of low and high concentration uric acid solution at the same time. The addition of uric acid with the concentrations from 1 pM to 0.5 mM resulted in the electrical conductance changes of up to 227 nS, and the response time turns out to be in the order of millisecond. The ZnO NW biosensor could easily detect as low as 1 pM of the uric acid with 14.7 nS of conductance increase, which implied that the sensitivity of the biosensor can be below the 1pM concentration.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Martin, Audrey Noreen
2006-01-01
Single Particle Aerosol Mass Spectrometry (SPAMS) was evaluated as a real-time detection technique for single particles of high explosives. Dual-polarity time-of-flight mass spectra were obtained for samples of 2,4,6-trinitrotoluene (TNT), 1,3,5-trinitro-1,3,5-triazinane (RDX), and pentaerythritol tetranitrate (PETN); peaks indicative of each compound were identified. Composite explosives, Comp B, Semtex 1A, and Semtex 1H were also analyzed, and peaks due to the explosive components of each sample were present in each spectrum. Mass spectral variability with laser fluence is discussed. The ability of the SPAMS system to identify explosive components in a single complex explosive particle (~1 pg) without the need formore » consumables is demonstrated. SPAMS was also applied to the detection of Chemical Warfare Agent (CWA) simulants in the liquid and vapor phases. Liquid simulants for sarin, cyclosarin, tabun, and VX were analyzed; peaks indicative of each simulant were identified. Vapor phase CWA simulants were adsorbed onto alumina, silica, Zeolite, activated carbon, and metal powders which were directly analyzed using SPAMS. The use of metal powders as adsorbent materials was especially useful in the analysis of triethyl phosphate (TEP), a VX stimulant, which was undetectable using SPAMS in the liquid phase. The capability of SPAMS to detect high explosives and CWA simulants using one set of operational conditions is established.« less
DOE Office of Scientific and Technical Information (OSTI.GOV)
Teo, Stephanie M.; Ofori-Okai, Benjamin K.; Werley, Christopher A.
Multidimensional spectroscopy at visible and infrared frequencies has opened a window into the transfer of energy and quantum coherences at ultrafast time scales. For these measurements to be performed in a manageable amount of time, one spectral axis is typically recorded in a single laser shot. An analogous rapid-scanning capability for THz measurements will unlock the multidimensional toolkit in this frequency range. Here, we first review the merits of existing single-shot THz schemes and discuss their potential in multidimensional THz spectroscopy. We then introduce improved experimental designs and noise suppression techniques for the two most promising methods: frequency-to-time encoding withmore » linear spectral interferometry and angle-to-time encoding with dual echelons. Both methods, each using electro-optic detection in the linear regime, were able to reproduce the THz temporal waveform acquired with a traditional scanning delay line. Although spectral interferometry had mediocre performance in terms of signal-to-noise, the dual echelon method was easily implemented and achieved the same level of signal-to-noise as the scanning delay line in only 4.5% of the laser pulses otherwise required (or 22 times faster). This reduction in acquisition time will compress day-long scans to hours and hence provides a practical technique for multidimensional THz measurements.« less
Teo, Stephanie M; Ofori-Okai, Benjamin K; Werley, Christopher A; Nelson, Keith A
2015-05-01
Multidimensional spectroscopy at visible and infrared frequencies has opened a window into the transfer of energy and quantum coherences at ultrafast time scales. For these measurements to be performed in a manageable amount of time, one spectral axis is typically recorded in a single laser shot. An analogous rapid-scanning capability for THz measurements will unlock the multidimensional toolkit in this frequency range. Here, we first review the merits of existing single-shot THz schemes and discuss their potential in multidimensional THz spectroscopy. We then introduce improved experimental designs and noise suppression techniques for the two most promising methods: frequency-to-time encoding with linear spectral interferometry and angle-to-time encoding with dual echelons. Both methods, each using electro-optic detection in the linear regime, were able to reproduce the THz temporal waveform acquired with a traditional scanning delay line. Although spectral interferometry had mediocre performance in terms of signal-to-noise, the dual echelon method was easily implemented and achieved the same level of signal-to-noise as the scanning delay line in only 4.5% of the laser pulses otherwise required (or 22 times faster). This reduction in acquisition time will compress day-long scans to hours and hence provides a practical technique for multidimensional THz measurements.
Loeschner, Katrin; Correia, Manuel; López Chaves, Carlos; Rokkjær, Inge; Sloth, Jens J
2018-01-01
This study investigated Chinese noodles for the presence of aluminium-containing nanoparticles by using inductively coupled plasma mass spectrometry in single particle mode (spICP-MS) after enzymatic digestion by α-amylase. The aluminium concentrations in the noodle samples, determined by conventional ICP-MS without or with the use of hydrofluoric acid for digestion, were 5.4 ± 1.9 µg/g and 10.1 ± 2.2 µg/g (N = 21), respectively. Aluminium-containing nanoparticles were detected by spICP-MS in all 21 samples. Depending on the assumed particle composition, Al 2 O 3 or Al 2 O 3 ∙2SiO 2 ∙2H 2 O, the median particle diameters were either below or above 100 nm, respectively. The minimum detectable particle diameter by spICP-MS was between 54 and 83 nm. The mass recovery of aluminium in the form of particles was between 5% and 18%. The presented work reports for the first time the detection of Al-containing particles in food by spICP-MS.
Ma, Xingyi; Sim, Sang Jun
2013-03-21
Even though DNA-based nanosensors have been demonstrated for quantitative detection of analytes and diseases, hybridization events have never been numerically investigated for further understanding of DNA mediated interactions. Here, we developed a nanoscale platform with well-designed capture and detection gold nanoprobes to precisely evaluate the hybridization events. The capture gold nanoprobes were mono-laid on glass and the detection probes were fabricated via a novel competitive conjugation method. The two kinds of probes combined in a suitable orientation following the hybridization with the target. We found that hybridization efficiency was markedly dependent on electrostatic interactions between DNA strands, which can be tailored by adjusting the salt concentration of the incubation solution. Due to the much lower stability of the double helix formed by mismatches, the hybridization efficiencies of single mismatched (MMT) and perfectly matched DNA (PMT) were different. Therefore, we obtained an optimized salt concentration that allowed for discrimination of MMT from PMT without stringent control of temperature or pH. The results indicated this to be an ultrasensitive and precise nanosensor for the diagnosis of genetic diseases.
Ho, Emmie N M; Kwok, W H; Wong, April S Y; Wan, Terence S M
2012-01-13
Quaternary ammonium drugs (QADs) are anticholinergic agents some of which are known to have been abused or misused in equine sports. A recent review of literature shows that the screening methods reported thus far for QADs mainly cover singly-charged QADs. Doubly-charged QADs are extremely polar substances which are difficult to be extracted and poorly retained on reversed-phase columns. It would be ideal if a comprehensive method can be developed which can detect both singly- and doubly-charged QADs. This paper describes an efficient liquid chromatography/tandem mass spectrometry (LC/MS/MS) method for the simultaneous detection and confirmation of 38 singly- and doubly-charged QADs at sub-parts-per-billion (ppb) to low-ppb levels in equine urine after solid-phase extraction. Quaternary ammonium drugs were extracted from equine urine by solid-phase extraction (SPE) using an ISOLUTE(®) CBA SPE column and analysed by LC/MS/MS in the positive electrospray ionisation mode. Separation of the 38 QADs was achieved on a polar group embedded C18 LC column with a mixture of aqueous ammonium formate (pH 3.0, 10 mM) and acetonitrile as the mobile phase. Detection and confirmation of the 38 QADs at sub-ppb to low-ppb levels in equine urine could be achieved within 16 min using selected reaction monitoring (SRM). Matrix interference of the target transitions at the expected retention times was not observed. Other method validation data, including precision and recovery, were acceptable. The method was successfully applied to the analyses of drug-administration samples. Copyright © 2011 Elsevier B.V. All rights reserved.
DOE Office of Scientific and Technical Information (OSTI.GOV)
Gao, Qing, E-mail: qing.gao.chance@gmail.com; Dong, Daoyi, E-mail: daoyidong@gmail.com; Petersen, Ian R., E-mail: i.r.petersen@gmai.com
The purpose of this paper is to solve the fault tolerant filtering and fault detection problem for a class of open quantum systems driven by a continuous-mode bosonic input field in single photon states when the systems are subject to stochastic faults. Optimal estimates of both the system observables and the fault process are simultaneously calculated and characterized by a set of coupled recursive quantum stochastic differential equations.
Wang, Jianchang; Li, Rui; Hu, Lianxia; Sun, Xiaoxia; Wang, Jinfeng; Li, Jing
2016-02-16
Food-borne disease caused by Salmonella has long been, and continues to be, an important global public health problem, necessitating rapid and accurate detection of Salmonella in food. Real time PCR is the most recently developed approach for Salmonella detection. Single primer isothermal amplification (SPIA), a novel gene amplification technique, has emerged as an attractive microbiological testing method. SPIA is performed under a constant temperature, eliminating the need for an expensive thermo-cycler. In addition, SPIA reactions can be accomplished in 30 min, faster than real time PCR that usually takes over 2h. We developed a quantitative fluorescence SPIA-based method for the detection of Salmonella. Using Salmonella Typhimurium genomic DNA as template and a primer targeting Salmonella invA gene, we showed the detection limit of SPIA was 2.0 × 10(1)fg DNA. Its successful amplification of different serotypic Salmonella genomic DNA but not non-Salmonella bacterial DNA demonstrated the specificity of SPIA. Furthermore, this method was validated with artificially contaminated beef. In conclusion, we showed high sensitivity and specificity of SPIA in the detection of Salmonella, comparable to real time PCR. In addition, SPIA is faster and more cost-effective (non-use of expensive cyclers), making it a potential alternative for field detection of Salmonella in resource-limited settings that are commonly encountered in developing countries. Copyright © 2015 Elsevier B.V. All rights reserved.
Comprehensive GMO detection using real-time PCR array: single-laboratory validation.
Mano, Junichi; Harada, Mioko; Takabatake, Reona; Furui, Satoshi; Kitta, Kazumi; Nakamura, Kosuke; Akiyama, Hiroshi; Teshima, Reiko; Noritake, Hiromichi; Hatano, Shuko; Futo, Satoshi; Minegishi, Yasutaka; Iizuka, Tayoshi
2012-01-01
We have developed a real-time PCR array method to comprehensively detect genetically modified (GM) organisms. In the method, genomic DNA extracted from an agricultural product is analyzed using various qualitative real-time PCR assays on a 96-well PCR plate, targeting for individual GM events, recombinant DNA (r-DNA) segments, taxon-specific DNAs, and donor organisms of the respective r-DNAs. In this article, we report the single-laboratory validation of both DNA extraction methods and component PCR assays constituting the real-time PCR array. We selected some DNA extraction methods for specified plant matrixes, i.e., maize flour, soybean flour, and ground canola seeds, then evaluated the DNA quantity, DNA fragmentation, and PCR inhibition of the resultant DNA extracts. For the component PCR assays, we evaluated the specificity and LOD. All DNA extraction methods and component PCR assays satisfied the criteria set on the basis of previous reports.
High-throughput, dual probe biological assays based on single molecule detection
Hollars, Christopher W [Brentwood, CA; Huser, Thomas R [Livermore, CA; Lane, Stephen M [Oakland, CA; Balhorn, Rodney L [Livermore, CA; Bakajin, Olgica [San Leandro, CA; Darrow, Christopher [Pleasanton, CA; Satcher, Jr., Joe H.
2006-07-11
A method and apparatus with the sensitivity to detect and identify single target molecules through the localization of dual, fluorescently labeled probe molecules. This can be accomplished through specific attachment of the taget to a surface or in a two-dimensional (2D) flowing fluid sheet having approximate dimensions of 0.5 .mu.m.times.100 .mu.m.times.100 .mu.m. A device using these methods would have 10.sup.3 10.sup.4 greater throughput than previous one-dimensional (1D) micro-stream devices having 1 .mu.m.sup.3 interrogation volumes and would for the first time allow immuno- and DNA assays at ultra-low (femtomolar) concentrations to be performed in short time periods (.about.10 minutes). The use of novel labels (such as metal or semiconductor nanoparticles) may be incorporated to further extend the sensitivity possibly into the attomolar range.
Histopathologic features of ovarian borderline tumors are not predictive of clinical outcome.
Avril, Stefanie; Hahn, Ellen; Specht, Katja; Hauptmann, Steffen; Höss, Cornelia; Kiechle, Marion; Höfler, Heinz; Schmalfeldt, Barbara
2012-12-01
Ovarian borderline tumors (BOTs) generally have an excellent prognosis, although recurrences and malignant transformation can occur. Our aim was to compare clinicopathologic features of BOT with clinical outcome. In seventy consecutive BOTs clinicopathologic parameters, tumor cell proliferation (Ki67) and in selected cases KRAS, BRAF and p53 mutational status were analyzed with recurrence-free and overall survival as the endpoints. Sixty-one (87%) patients presented with FIGO stage I, 3 stage II, and 6 stage III. Thirty-four patients had serous and 36 mucinous BOT (30 intestinal and 6 endocervical subtypes). Non-invasive peritoneal implants occurred in 9 patients, and no invasive implants were observed. Recurrence-free and overall survival rates were 91% and 99%, respectively, at a mean follow-up of 63 months. Disease recurrence occurred in 6 cases (all FIGO stage I) including 3 serous, 1 mucinous-intestinal, and 2 mucinous-endocervical subtypes. Mean time to recurrence was 27 months (range 8-68). The recurrence rate following fertility-conserving surgery was 31% (5/16) compared to 2% (1/54) after bilateral salpingo-oophorectomy. Neither peritoneal implants (9/70), micropapillary pattern (2/34), microinvasion (4/70), nor increased tumor cell proliferation was associated with a higher recurrence rate. The frequency of KRAS or BRAF mutations was 50% (3/6 recurrences and 3/6 controls; 4 KRAS, 2 BRAF mutations). No p53 mutations (0/12) were detected in primary or recurrent BOTs. Histopathologic parameters were not predictive of BOT recurrence including previously suggested risk factors such as micropapillary pattern and microinvasion. However, fertility-conserving surgery and incomplete surgical staging were associated with a higher risk for recurrence. Copyright © 2012 Elsevier B.V. All rights reserved.
Prototype Test Results for the Single Photon Detection SLR2000 Satellite Laser Ranging System
NASA Technical Reports Server (NTRS)
Zagwodzki, Thomas W.; McGarry, Jan F.; Degnan, John J.; Cheek, Jack W.; Dunn, Peter J.; Patterson, Don; Donovan, Howard
2004-01-01
NASA's aging Satellite Laser Ranging (SLR) network is scheduled to be replaced over the next few years with a fully automated single photon detection system. A prototype of this new system, called SLR2000, is currently undergoing field trials at the Goddard Space Flight Center in Greenbelt, Maryland to evaluate photon counting techniques and determine system hardware, software, and control algorithm performance levels and limitations. Newly developed diode pumped microchip lasers and quadrant microchannel plate-based photomultiplier tubes have enabled the development of this high repetition rate single photon detection SLR system. The SLR2000 receiver threshold is set at the single photoelectron (pe) level but tracks satellites with an average signal level typically much less than 1 pe. The 2 kHz laser fire rate aids in satellite acquisition and tracking and will enable closed loop tracking by accumulating single photon count statistics in a quadrant detector and using this information to correct for pointing errors. Laser transmitter beamwidths of 10 arcseconds (FWHM) or less are currently being used to maintain an adequate signal level for tracking while the receiver field of view (FOV) has been opened to 40 arcseconds to accommodate point ahead/look behind angular offsets. In the near future, the laser transmitter point ahead will be controlled by a pair of Risley prisms. This will allow the telescope to point behind and enable closure of the receiver FOV to roughly match the transmitter beam divergence. Bandpass filters (BPF) are removed for night tracking operations while 0.2 nm or 1 nm filters are used during daylight operation. Both day and night laser tracking of Low Earth Orbit (LEO) satellites has been achieved with a laser transmitter energy of only 65 microjoules per pulse. Satellite tracking is presently limited to LEO satellites until the brassboard laser transmitter can be upgraded or replaced. Simultaneous tracks have also been observed with NASA s
Dittfeld, Anna; Gwizdek, Katarzyna; Jagielski, Paweł; Brzęk, Joanna; Ziora, Katarzyna
2017-12-30
The following article presents the relationship between vegetarianism and orthorexia nervosa (ON). Vegetarianism is an ideology and a way of life that aims at minimizing animal exploitation. A vegetarian diet excludes the consumption of meat together with other animal derived products. According to scientists, orthorexia nervosa is considered to be a new, yet unclassified eating disorder. It involves introducing dietary restrictions by individuals who feel a desire to improve their health status by healthy eating. The study involved 2,611 participants, namely 1,346 vegetarians and 1,265 non-vegetarians. The research questionnaire consisted of general personal and anthropometric characteristics, the BOT(Bratman Test for Orthorexia) and questions evaluating the participants' attitude towards nutrition. Based on the obtained results, health food fanaticism is more specific to vegetarians than non-vegetarians. The risk for orthorexia nervosa decreases with age and diet duration. The biggest number of health food fanatics was found in the group of lacto-vegetarians, a lower number among ovo-vegetarians and lacto-ovo-vegetarians, and the smallest number was observed in the vegan group. Also, vegetarians were reported to have dietary consultations as frequently as non-vegetarians. Very few studies can be found on the relationship between orthorexia nervosa and vegetarianism. Some scientists believe that vegetarians are particularly prone to orthorexia nervosa. In addition, it has been suggested by other researchers that vegetarianism can be used to mask eating disorders, as it allows these affected individuals to avoid certain products or situations related to food. The direction of cause and effect cannot be determined.
LABRADOR: a learning autonomous behavior-based robot for adaptive detection and object retrieval
NASA Astrophysics Data System (ADS)
Yamauchi, Brian; Moseley, Mark; Brookshire, Jonathan
2013-01-01
As part of the TARDEC-funded CANINE (Cooperative Autonomous Navigation in a Networked Environment) Program, iRobot developed LABRADOR (Learning Autonomous Behavior-based Robot for Adaptive Detection and Object Retrieval). LABRADOR was based on the rugged, man-portable, iRobot PackBot unmanned ground vehicle (UGV) equipped with an explosives ordnance disposal (EOD) manipulator arm and a custom gripper. For LABRADOR, we developed a vision-based object learning and recognition system that combined a TLD (track-learn-detect) filter based on object shape features with a color-histogram-based object detector. Our vision system was able to learn in real-time to recognize objects presented to the robot. We also implemented a waypoint navigation system based on fused GPS, IMU (inertial measurement unit), and odometry data. We used this navigation capability to implement autonomous behaviors capable of searching a specified area using a variety of robust coverage strategies - including outward spiral, random bounce, random waypoint, and perimeter following behaviors. While the full system was not integrated in time to compete in the CANINE competition event, we developed useful perception, navigation, and behavior capabilities that may be applied to future autonomous robot systems.
Recent advances in single-molecule detection on micro- and nano-fluidic devices.
Liu, Chang; Qu, Yueyang; Luo, Yong; Fang, Ning
2011-11-01
Single-molecule detection (SMD) allows static and dynamic heterogeneities from seemingly equal molecules to be revealed in the studies of molecular structures and intra- and inter-molecular interactions. Micro- and nanometer-sized structures, including channels, chambers, droplets, etc., in microfluidic and nanofluidic devices allow diffusion-controlled reactions to be accelerated and provide high signal-to-noise ratio for optical signals. These two active research frontiers have been combined to provide unprecedented capabilities for chemical and biological studies. This review summarizes the advances of SMD performed on microfluidic and nanofluidic devices published in the past five years. The latest developments on optical SMD methods, microfluidic SMD platforms, and on-chip SMD applications are discussed herein and future development directions are also envisioned. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Single-view phase retrieval of an extended sample by exploiting edge detection and sparsity
Tripathi, Ashish; McNulty, Ian; Munson, Todd; ...
2016-10-14
We propose a new approach to robustly retrieve the exit wave of an extended sample from its coherent diffraction pattern by exploiting sparsity of the sample's edges. This approach enables imaging of an extended sample with a single view, without ptychography. We introduce nonlinear optimization methods that promote sparsity, and we derive update rules to robustly recover the sample's exit wave. We test these methods on simulated samples by varying the sparsity of the edge-detected representation of the exit wave. Finally, our tests illustrate the strengths and limitations of the proposed method in imaging extended samples.
Llop, Pablo; Bonaterra, Anna; Peñalver, Javier; López, María M.
2000-01-01
A novel method, which involves a nested PCR in a single closed tube, was developed for the sensitive detection of Erwinia amylovora in plant material. The external and internal primer pairs used had different annealing temperatures and directed the amplification of a specific DNA fragment from plasmid pEA29. The procedure involved two consecutive PCRs, the first of which was performed at a higher annealing temperature that allowed amplification only by the external primer pair. Using pure cultures of E. amylovora, the sensitivity of the nested PCR in one tube was similar to that of a standard nested PCR in two tubes. The specificity and sensitivity were greater than those of standard PCR procedures that used a single primer pair. The presence of inhibitors in plant material, very common in E. amylovora hosts, is overcome with this system in combination with a simple DNA extraction protocol because it eliminates many of the inhibitory compounds. In addition, it needs a very small sample volume (1 μl of DNA extracted). With 83 samples of naturally infected material, this method achieved better results than any other PCR technique: standard PCR detected 55% of positive samples, two-tube nested PCR detected 71% of positive samples, and nested PCR in a single closed tube detected 78% of positive samples. When analyzing asymptomatic plant material, the number of positive samples detected by the developed nested PCR was also the highest, compared with the PCR protocols indicated previously (17, 20, and 25% of 251 samples analyzed, respectively). This method is proposed for the detection of endophytic and epiphytic populations of E. amylovora in epidemiological studies and for routine use in quarantine surveys, due to its high sensitivity, specificity, speed, and simplicity. PMID:10788384
Breadth-First Search-Based Single-Phase Algorithms for Bridge Detection in Wireless Sensor Networks
Akram, Vahid Khalilpour; Dagdeviren, Orhan
2013-01-01
Wireless sensor networks (WSNs) are promising technologies for exploring harsh environments, such as oceans, wild forests, volcanic regions and outer space. Since sensor nodes may have limited transmission range, application packets may be transmitted by multi-hop communication. Thus, connectivity is a very important issue. A bridge is a critical edge whose removal breaks the connectivity of the network. Hence, it is crucial to detect bridges and take preventions. Since sensor nodes are battery-powered, services running on nodes should consume low energy. In this paper, we propose energy-efficient and distributed bridge detection algorithms for WSNs. Our algorithms run single phase and they are integrated with the Breadth-First Search (BFS) algorithm, which is a popular routing algorithm. Our first algorithm is an extended version of Milic's algorithm, which is designed to reduce the message length. Our second algorithm is novel and uses ancestral knowledge to detect bridges. We explain the operation of the algorithms, analyze their proof of correctness, message, time, space and computational complexities. To evaluate practical importance, we provide testbed experiments and extensive simulations. We show that our proposed algorithms provide less resource consumption, and the energy savings of our algorithms are up by 5.5-times. PMID:23845930
Single molecule detection with graphene and other two-dimensional materials: nanopores and beyond
Arjmandi-Tash, Hadi; Belyaeva, Liubov A.
2016-01-01
Graphene and other two dimensional (2D) materials are currently integrated into nanoscaled devices that may – one day – sequence genomes. The challenge to solve is conceptually straightforward: cut a sheet out of a 2D material and use the edge of the sheet to scan an unfolded biomolecule from head to tail. As the scan proceeds – and because 2D materials are atomically thin – the information provided by the edge might be used to identify different segments – ideally single nucleotides – in the biomolecular strand. So far, the most efficient approach was to drill a nano-sized pore in the sheet and use this pore as a channel to guide and detect individual molecules by measuring the electrochemical ionic current. Nanoscaled gaps between two electrodes in 2D materials recently emerged as powerful alternatives to nanopores. This article reviews the current status and prospects of integrating 2D materials in nanopores, nanogaps and similar devices for single molecule biosensing applications. We discuss the pros and cons, the challenges, and the latest achievements in the field. To achieve high-throughput sequencing with 2D materials, interdisciplinary research is essential. PMID:26612268
Hamzah, Zulhainan; Petmitr, Songsak; Mungthin, Mathirut; Leelayoova, Saovanee; Chavalitshewinkoon-Petmitr, Porntip
2006-01-01
A single-round PCR assay was developed for detection and differential diagnosis of the three Entamoeba species found in humans, Entamoeba moshkovskii, Entamoeba histolytica, and Entamoeba dispar, that are morphologically identical as both cysts and trophozoites. A conserved forward primer was derived from the middle of the small-subunit rRNA gene, and reverse primers were designed from signature sequences specific to each of these three Entamoeba species. PCR generates a 166-bp product with E. histolytica DNA, a 752-bp product with E. dispar DNA, and a 580-bp product with E. moshkovskii DNA. Thirty clinical specimens were examined, and the species present were successfully detected and differentiated using this assay. It was possible to detect as little as 10 pg of E. moshkovskii and E. histolytica DNA, while for E. dispar the sensitivity was about 20 pg of DNA. Testing with DNA from different pathogens, including bacteria and other protozoa, confirmed the high specificity of the assay. We propose the use of this PCR assay as an accurate, rapid, and effective diagnostic method for the detection and discrimination of these three morphologically indistinguishable Entamoeba species in both routine diagnosis of amoebiasis and epidemiological surveys. PMID:16954247
Pradhan, Somarpita; Chaudhuri, Partha Roy
2015-07-10
We experimentally demonstrate single-mode optical-fiber-beam-deflection configuration for weak magnetic-field-detection using an optimized (low coercive-field) composition of cobalt-doped nickel ferrite nanoparticles. Devising a fiber-double-slit type experiment, we measure the surrounding magnetic field through precisely measuring interference-fringe yielding a minimum detectable field ∼100 mT and we procure magnetization data of the sample that fairly predicts SQUID measurement. To improve sensitivity, we incorporate etched single-mode fiber in double-slit arrangement and recorded a minimum detectable field, ∼30 mT. To further improve, we redefine the experiment as modulating fiber-to-fiber light-transmission and demonstrate the minimum field as 2.0 mT. The device will be uniquely suited for electrical or otherwise hazardous environments.
Banyard, Ashley C; Johnson, N; Voller, K; Hicks, D; Nunez, A; Hartley, M; Fooks, A R
2009-01-01
In August 2007, European bat lyssavirus type 2 (EBLV-2) was isolated from a Daubenton's bat found at Stokesay Castle. In September 2008, another bat from the same vicinity of Stokesay Castle also tested positive for EBLV-2. This is the first occurrence of repeated detection of EBLV-2 from a single site. Here, we report the detection of low levels of viral RNA in various bat organs by qRT-PCR and detection of viral antigen by immunohistochemistry. We also report sequence data from both cases and compare data with those derived from other EBLV-2 isolations in the UK.
Liang, L; Jin, Y X; Zhu, X Q; Zhou, F L; Yang, Y
2018-05-15
Real-time detection and monitoring of the drug resistance of single cells have important significance in clinical diagnosis and therapy. Traditional methods operate a number of times for each individual concentration, and innovation is required for the design of more simple and efficient manipulation platforms with necessary higher sensitivity. Here, we have developed a novel diffused total internal reflection (TIR) method to perform drug metabolism and cytotoxicity analysis of trapped myeloid leukemia cells. Molm-13 cells, a type of acute myeloid leukemia cell, were chosen and injected into the device and fittingly captured by cell traps. Differing from previous studies, a series of different concentrations of azelaic acid (AZA) drug could be used from 0 mM to 50 mM through convection and diffusion processes in a single chip, with each concentration region featuring 50 cells, with a total of 549 cell trapping units. Thanks to the high sensitivity of the TIR method, only cells with the same drug concentration could be illuminated in the detection process. By adjusting the incident angle, we could exactly detect and monitor the drug resistance of the cells using different drug concentrations and the experimental resolution of the drug concentration was as small as 5 mM. Images of the membrane integrity and morphology of the cells in the bright field were measured and we also monitored the cell viabilities in the dark field over 2 hours. The effects of AZA on the Molm-13 cells were explored in different concentrations at the single cell level. Compared with the results of the traditional MTT assay method, the experimental results are more simple and accurate. A cell death of 5% at an AZA concentration of 5 mM was observed after 30 minutes, while a concentration of 40 mM corresponded to a 98% cell death. The designed method in this study provides a novel toolkit to control and monitor drug resistance at the single cell level more easily with higher sensitivity and we
Towards Single-Shot Detection of Bacterial Endospores via Coherent Raman Spectroscopy
NASA Astrophysics Data System (ADS)
Pestov, Dmitry; Wang, Xi; Ariunbold, Gombojav; Murawski, Robert; Sautenkov, Vladimir; Sokolov, Alexei; Scully, Marlan
2007-10-01
Recent advances in coherent anti-Stokes Raman scattering (CARS) spectroscopy hold exciting promise to make the most out of now readily available ultrafast laser sources. Techniques have been devised to mitigate the nonresonant four-wave-mixing in favor of informative Raman-resonant signal. In particular, a hybrid technique for CARS (see Science 316, 265 (2007)) brings together the advantages of coherent broadband pump-Stokes excitation of molecular vibrations and their time-delayed but frequency-resolved probing via a spectrally narrowed and shaped laser pulse. We apply this technique to the problem of real-time detection of warfare bioagents and report single-shot acquisition of a distinct CARS spectrum from a small volume of B. subtilis endospores (˜10^4 spores), a harmless surrogate for B. anthracis. We study the dependence of the CARS signal on the energy of the ultrashort preparation pulses and find the limit on the pulse energy fluence (˜0.2 J/cm^2), imposed by the laser-induced damage of the spores.
NASA Astrophysics Data System (ADS)
Keifer, David Z.; Alexander, Andrew W.; Jarrold, Martin F.
2017-03-01
Spontaneous mass and charge losses from individual multi-megadalton ions have been observed with charge detection mass spectrometry (CDMS) by trapping single hepatitis B virus (HBV) capsids for 3 s. Gradual increases in the oscillation frequency of single ions in the ion trap are attributed mainly to mass loss (probably solvent, water, and/or salt). The total mass lost during the 3 s trapping period peaks at around 20 kDa for 4 MDa HBV T = 4 capsids. Discrete frequency drops punctuate the gradual increases in the oscillation frequencies. The drops are attributed to a sudden loss of charge. In most cases a single positive charge is lost along with some mass (on average around 1000 Da). Charge loss occurs for over 40% of the trapped ions. It usually occurs near the beginning of the trapping event, and it occurs preferentially in regions of the trap with strong electric fields, indicating that external electric fields promote charge loss. This process may contribute to the decrease in m/ z resolution that often occurs with megadalton ions.
Zhu, Mingyue; Zhang, Jing; Yi, Xingwen; Ying, Hao; Li, Xiang; Luo, Ming; Song, Yingxiong; Huang, Xiatao; Qiu, Kun
2018-03-19
We present the design and optimization of the optical single side-band (SSB) Nyquist four-level pulse amplitude modulation (PAM-4) transmission using dual-drive Mach-Zehnder modulator (DDMZM)modulation and direct detection (DD), aiming at the C-band cost-effective, high-speed and long-distance transmission. At the transmitter, the laser line width should be small to avoid the phase noise to amplitude noise conversion and equalization-enhanced phase noise due to the large chromatic dispersion (CD). The optical SSB signal is generated after optimizing the optical modulation index (OMI) and hence the minimum phase condition which is required by the Kramers-Kronig (KK) receiver can also be satisfied. At the receiver, a simple AC-coupled photodiode (PD) is used and a virtual carrier is added for the KK operation to alleviate the signal-to-signal beating interference (SSBI).A Volterra filter (VF) is cascaded for remaining nonlinearities mitigation. When the fiber nonlinearity becomes significant, we elect to use an optical band-pass filter with offset filtering. It can suppress the simulated Brillouin scattering and the conjugated distortion by filtering out the imaging frequency components. With our design and optimization, we achieve single-channel, single polarization 102.4-Gb/s Nyquist PAM-4 over 800-km standard single-mode fiber (SSMF).
Blom, H; Gösch, M
2004-04-01
The past few years we have witnessed a tremendous surge of interest in so-called array-based miniaturised analytical systems due to their value as extremely powerful tools for high-throughput sequence analysis, drug discovery and development, and diagnostic tests in medicine (see articles in Issue 1). Terminologies that have been used to describe these array-based bioscience systems include (but are not limited to): DNA-chip, microarrays, microchip, biochip, DNA-microarrays and genome chip. Potential technological benefits of introducing these miniaturised analytical systems include improved accuracy, multiplexing, lower sample and reagent consumption, disposability, and decreased analysis times, just to mention a few examples. Among the many alternative principles of detection-analysis (e.g.chemiluminescence, electroluminescence and conductivity), fluorescence-based techniques are widely used, examples being fluorescence resonance energy transfer, fluorescence quenching, fluorescence polarisation, time-resolved fluorescence, and fluorescence fluctuation spectroscopy (see articles in Issue 11). Time-dependent fluctuations of fluorescent biomolecules with different molecular properties, like molecular weight, translational and rotational diffusion time, colour and lifetime, potentially provide all the kinetic and thermodynamic information required in analysing complex interactions. In this mini-review article, we present recent extensions aimed to implement parallel laser excitation and parallel fluorescence detection that can lead to even further increase in throughput in miniaturised array-based analytical systems. We also report on developments and characterisations of multiplexing extension that allow multifocal laser excitation together with matched parallel fluorescence detection for parallel confocal dynamical fluorescence fluctuation studies at the single biomolecule level.
Schramm, Elisabeth; Kürten, Andreas; Hölzer, Jasper; Mitschke, Stefan; Mühlberger, Fabian; Sklorz, Martin; Wieser, Jochen; Ulrich, Andreas; Pütz, Michael; Schulte-Ladbeck, Rasmus; Schultze, Rainer; Curtius, Joachim; Borrmann, Stephan; Zimmermann, Ralf
2009-06-01
An in-house-built ion trap mass spectrometer combined with a soft ionization source has been set up and tested. As ionization source, an electron beam pumped vacuum UV (VUV) excimer lamp (EBEL) was used for single-photon ionization. It was shown that soft ionization allows the reduction of fragmentation of the target analytes and the suppression of most matrix components. Therefore, the combination of photon ionization with the tandem mass spectrometry (MS/MS) capability of an ion trap yields a powerful tool for molecular ion peak detection and identification of organic trace compounds in complex matrixes. This setup was successfully tested for two different applications. The first one is the detection of security-relevant substances like explosives, narcotics, and chemical warfare agents. One test substance from each of these groups was chosen and detected successfully with single photon ionization ion trap mass spectrometry (SPI-ITMS) MS/MS measurements. Additionally, first tests were performed, demonstrating that this method is not influenced by matrix compounds. The second field of application is the detection of process gases. Here, exhaust gas from coffee roasting was analyzed in real time, and some of its compounds were identified using MS/MS studies.
A single-shot nanosecond neutron pulsed technique for the detection of fissile materials
NASA Astrophysics Data System (ADS)
Gribkov, V.; Miklaszewski, R. A.; Chernyshova, M.; Scholz, M.; Prokopovicz, R.; Tomaszewski, K.; Drozdowicz, K.; Wiacek, U.; Gabanska, B.; Dworak, D.; Pytel, K.; Zawadka, A.
2012-07-01
A novel technique with the potential of detecting hidden fissile materials is presented utilizing the interaction of a single powerful and nanosecond wide neutron pulse with matter. The experimental system is based on a Dense Plasma Focus (DPF) device as a neutron source generating pulses of almost mono-energetic 2.45 MeV and/or 14.0 MeV neutrons, a few nanoseconds in width. Fissile materials, consisting of heavy nuclei, are detected utilizing two signatures: firstly by measuring those secondary fission neutrons which are faster than the elastically scattered 2.45 MeV neutrons of the D-D reaction in the DPF; secondly by measuring the pulses of the slower secondary fission neutrons following the pulse of the fast 14 MeV neutrons from the D-T reaction. In both cases it is important to compare the measured spectrum of the fission neutrons induced by the 2.45 MeV or 14 MeV neutron pulse of the DPF with theoretical spectra obtained by mathematical simulation. Therefore, results of numerical modelling of the proposed system, using the MCNP5 and the FLUKA codes are presented and compared with experimental data.
Robertson, Neil M; Hizir, Mustafa Salih; Balcioglu, Mustafa; Wang, Rui; Yavuz, Mustafa Selman; Yumak, Hasan; Ozturk, Birol; Sheng, Jia; Yigit, Mehmet V
2015-09-15
In this study we have reported our efforts to address some of the challenges in the detection of miRNAs using water-soluble graphene oxide and DNA nanoassemblies. Purposefully inserting mismatches at specific positions in our DNA (probe) strands shows increasing specificity against our target miRNA, miR-10b, over miR-10a which varies by only a single nucleotide. This increased specificity came at a loss of signal intensity within the system, but we demonstrated that this could be addressed with the use of DNase I, an endonuclease capable of cleaving the DNA strands of the RNA/DNA heteroduplex and recycling the RNA target to hybridize to another probe strand. As we previously demonstrated, this enzymatic signal also comes with an inherent activity of the enzyme on the surface-adsorbed probe strands. To remove this activity of DNase I and the steady nonspecific increase in the fluorescence signal without compromising the recovered signal, we attached a thermoresponsive PEGMA polymer (poly(ethylene glycol) methyl ether methacrylate) to nGO. This smart polymer is able to shield the probes adsorbed on the nGO surface from the DNase I activity and is capable of tuning the detection capacity of the nGO nanoassembly with a thermoswitch at 39 °C. By utilizing probes with multiple mismatches, DNase I cleavage of the DNA probe strands, and the attachment of PEGMA polymers to graphene oxide to block undesired DNase I activity, we were able to detect miR-10b from liquid biopsy mimics and breast cancer cell lines. Overall we have reported our efforts to improve the specificity, increase the sensitivity, and eliminate the undesired enzymatic activity of DNase I on surface-adsorbed probes for miR-10b detection using water-soluble graphene nanodevices. Even though we have demonstrated only the discrimination of miR-10b from miR-10a, our approach can be extended to other short RNA molecules which differ by a single nucleotide.
Precise detection of de novo single nucleotide variants in human genomes.
Gómez-Romero, Laura; Palacios-Flores, Kim; Reyes, José; García, Delfino; Boege, Margareta; Dávila, Guillermo; Flores, Margarita; Schatz, Michael C; Palacios, Rafael
2018-05-22
The precise determination of de novo genetic variants has enormous implications across different fields of biology and medicine, particularly personalized medicine. Currently, de novo variations are identified by mapping sample reads from a parent-offspring trio to a reference genome, allowing for a certain degree of differences. While widely used, this approach often introduces false-positive (FP) results due to misaligned reads and mischaracterized sequencing errors. In a previous study, we developed an alternative approach to accurately identify single nucleotide variants (SNVs) using only perfect matches. However, this approach could be applied only to haploid regions of the genome and was computationally intensive. In this study, we present a unique approach, coverage-based single nucleotide variant identification (COBASI), which allows the exploration of the entire genome using second-generation short sequence reads without extensive computing requirements. COBASI identifies SNVs using changes in coverage of exactly matching unique substrings, and is particularly suited for pinpointing de novo SNVs. Unlike other approaches that require population frequencies across hundreds of samples to filter out any methodological biases, COBASI can be applied to detect de novo SNVs within isolated families. We demonstrate this capability through extensive simulation studies and by studying a parent-offspring trio we sequenced using short reads. Experimental validation of all 58 candidate de novo SNVs and a selection of non-de novo SNVs found in the trio confirmed zero FP calls. COBASI is available as open source at https://github.com/Laura-Gomez/COBASI for any researcher to use. Copyright © 2018 the Author(s). Published by PNAS.
Fennell, John F.; Hamaguchi, Hitoshi; Yoon, Bora; Swager, Timothy M.
2017-01-01
Chemical warfare agents (CWA) continue to present a threat to civilian populations and military personnel in operational areas all over the world. Reliable measurements of CWAs are critical to contamination detection, avoidance, and remediation. The current deployed systems in United States and foreign militaries, as well as those in the private sector offer accurate detection of CWAs, but are still limited by size, portability and fabrication cost. Herein, we report a chemiresistive CWA sensor using single-walled carbon nanotubes (SWCNTs) wrapped with poly(3,4-ethylenedioxythiophene) (PEDOT) derivatives. We demonstrate that a pendant hexafluoroisopropanol group on the polymer that enhances sensitivity to a nerve agent mimic, dimethyl methylphosphonate, in both nitrogen and air environments to concentrations as low as 5 ppm and 11 ppm, respectively. Additionally, these PEDOT/SWCNT derivative sensor systems experience negligible device performance over the course of two weeks under ambient conditions. PMID:28452929
Fennell, John F; Hamaguchi, Hitoshi; Yoon, Bora; Swager, Timothy M
2017-04-28
Chemical warfare agents (CWA) continue to present a threat to civilian populations and military personnel in operational areas all over the world. Reliable measurements of CWAs are critical to contamination detection, avoidance, and remediation. The current deployed systems in United States and foreign militaries, as well as those in the private sector offer accurate detection of CWAs, but are still limited by size, portability and fabrication cost. Herein, we report a chemiresistive CWA sensor using single-walled carbon nanotubes (SWCNTs) wrapped with poly(3,4-ethylenedioxythiophene) (PEDOT) derivatives. We demonstrate that a pendant hexafluoroisopropanol group on the polymer that enhances sensitivity to a nerve agent mimic, dimethyl methylphosphonate, in both nitrogen and air environments to concentrations as low as 5 ppm and 11 ppm, respectively. Additionally, these PEDOT/SWCNT derivative sensor systems experience negligible device performance over the course of two weeks under ambient conditions.
First detection of nonflare microwave emissions from the coronae of single late-type dwarf stars
NASA Technical Reports Server (NTRS)
Gary, D. E.; Linsky, J. L.
1981-01-01
Results are presented of a search for nonflare microwave radiation from the coronae of nearby late-type dwarf stars comparable to the sun: single stars without evidence for either a large wind or circumstellar envelope. The observing program consisted of flux measurements of six stars over a 24-h period with the VLA in the C configuration at a wavelength of 6 cm with 50 MHz bandwidth. Positive detections at 6 cm were made for Chi 1 Ori (0.6 mJy) and the flare star UV Cet (1.55 mJy), and upper limits were obtained for the stars Pi 1 UMa, Xi Boo A, 70 Oph A and Epsilon Eri. It is suggested that Chi 1 Ori, and possibly UV Cet, represent the first detected members of a new class of radio sources which are driven by gyroresonance emission, i.e. cyclotron emission from nonrelativistic Maxwellian electrons.
Ahberg, Christian D.; Manz, Andreas; Neuzil, Pavel
2015-01-01
Since its invention in 1985 the polymerase chain reaction (PCR) has become a well-established method for amplification and detection of segments of double-stranded DNA. Incorporation of fluorogenic probe or DNA intercalating dyes (such as SYBR Green) into the PCR mixture allowed real-time reaction monitoring and extraction of quantitative information (qPCR). Probes with different excitation spectra enable multiplex qPCR of several DNA segments using multi-channel optical detection systems. Here we show multiplex qPCR using an economical EvaGreen-based system with single optical channel detection. Previously reported non quantitative multiplex real-time PCR techniques based on intercalating dyes were conducted once the PCR is completed by performing melting curve analysis (MCA). The technique presented in this paper is both qualitative and quantitative as it provides information about the presence of multiple DNA strands as well as the number of starting copies in the tested sample. Besides important internal control, multiplex qPCR also allows detecting concentrations of more than one DNA strand within the same sample. Detection of the avian influenza virus H7N9 by PCR is a well established method. Multiplex qPCR greatly enhances its specificity as it is capable of distinguishing both haemagglutinin (HA) and neuraminidase (NA) genes as well as their ratio. PMID:26088868
Single-pipetting microfluidic assay device for rapid detection of Salmonella from poultry package.
Fronczek, Christopher F; You, David J; Yoon, Jeong-Yeol
2013-02-15
A direct, sensitive, near-real-time, handheld optical immunoassay device was developed to detect Salmonella typhimurium in the naturally occurring liquid from fresh poultry packages (hereafter "chicken matrix"), with just single pipetting of sample (i.e., no filtration, culturing and/or isolation, thus reducing the assay time and the error associated with them). Carboxylated, polystyrene microparticles were covalently conjugated with anti-Salmonella, and the immunoagglutination due to the presence of Salmonella was detected by reading the Mie scatter signals from the microfluidic channels using a handheld device. The presence of chicken matrix did not affect the light scatter signal, since the optical parameters (particle size d, wavelength of incident light λ and scatter angle θ) were optimized to minimize the effect of sample matrix (animal tissues and blood proteins, etc.). The sample was loaded into a microfluidic chip that was split into two channels, one pre-loaded with vacuum-dried, antibody-conjugated particles and the other with vacuum-dried, bovine serum albumin-conjugated particles. This eliminated the need for a separate negative control, effectively minimizing chip-to-chip and sample-to-sample variations. Particles and the sample were diffused in-channel through chemical agitation by Tween 80, also vacuum-dried within the microchannels. Sequential mixing of the sample to the reagents under a strict laminar flow condition synergistically improved the reproducibility and linearity of the assay. In addition, dried particles were shown to successfully detect lower Salmonella concentrations for up to 8 weeks. The handheld device contains simplified circuitry eliminating unnecessary adjustment stages, providing a stable signal, thus maximizing sensitivity. Total assay time was 10 min, and the detection limit 10 CFU mL(-1) was observed in all matrices, demonstrating the suitability of this device for field assays. Copyright © 2012 Elsevier B.V. All rights
Laparoendoscopic Single-Site Sentinel Lymph Node Detection in Endometrial Cancer.
Demirayak, Gökhan; Comba, Cihan; Özdemir, İsa Aykut
2017-11-13
To demonstrate the feasibility of sentinel lymph node (SLN) biopsy using a laparoendoscopic single-site (LESS) approach in endometrial cancer (EC). A step-by-step video demonstration of the surgical procedure (Canadian Task Force Classification III). The satisfaction of patients who undergo LESS hysterectomy is greater than that reported by patients who undergo multiport laparoscopic hysterectomy, owing to better cosmesis and reduced postoperative analgesic requirements [1]. SLN biopsy is associated with significantly lower estimated blood loss, shorter operation time, and less morbidity compared with systematic lymphadenectomy [2]. LESS surgery can be more feasible and safer with the use of SLN biopsy compared with complete lymphadenectomy in patients with early-stage EC. This 69-year-old woman with grade 2 endometrioid EC underwent SLN mapping followed by LESS SLN biopsy, total hysterectomy, and bilateral salpingo-oophorectomy. Before the umbilical incision was made, 1.25 mg/mL of indocyanine green was injected into the cervical stroma at the 3 o'clock and 9 o'clock positions to both deep and superficial levels. A 10-mm 30° standard-length optical camera for near-infrared fluorescence imaging was used. The total operative time was 75 minutes, and the estimated blood loss was 20 mL. SLNs were detected bilaterally between proximal parts of the external iliac arteries and veins. After SLN resection, total hysterectomy and bilateral salpingo-oophorectomy were performed. No postoperative complications occurred. The patient was discharged at 30 hours after surgery. In the final pathology, stage 1A G2 EC was detected. LESS SLN biopsy and TLH-BSO is a feasible procedure and sentinel lymph node concept may increase the use of LESS in EC. Copyright © 2017 American Association of Gynecologic Laparoscopists. Published by Elsevier Inc. All rights reserved.
Integrated long-range UAV/UGV collaborative target tracking
NASA Astrophysics Data System (ADS)
Moseley, Mark B.; Grocholsky, Benjamin P.; Cheung, Carol; Singh, Sanjiv
2009-05-01
Coordinated operations between unmanned air and ground assets allow leveraging of multi-domain sensing and increase opportunities for improving line of sight communications. While numerous military missions would benefit from coordinated UAV-UGV operations, foundational capabilities that integrate stove-piped tactical systems and share available sensor data are required and not yet available. iRobot, AeroVironment, and Carnegie Mellon University are working together, partially SBIR-funded through ARDEC's small unit network lethality initiative, to develop collaborative capabilities for surveillance, targeting, and improved communications based on PackBot UGV and Raven UAV platforms. We integrate newly available technologies into computational, vision, and communications payloads and develop sensing algorithms to support vision-based target tracking. We first simulated and then applied onto real tactical platforms an implementation of Decentralized Data Fusion, a novel technique for fusing track estimates from PackBot and Raven platforms for a moving target in an open environment. In addition, system integration with AeroVironment's Digital Data Link onto both air and ground platforms has extended our capabilities in communications range to operate the PackBot as well as in increased video and data throughput. The system is brought together through a unified Operator Control Unit (OCU) for the PackBot and Raven that provides simultaneous waypoint navigation and traditional teleoperation. We also present several recent capability accomplishments toward PackBot-Raven coordinated operations, including single OCU display design and operation, early target track results, and Digital Data Link integration efforts, as well as our near-term capability goals.
Nanopore arrays in a silicon membrane for parallel single-molecule detection: fabrication
NASA Astrophysics Data System (ADS)
Schmidt, Torsten; Zhang, Miao; Sychugov, Ilya; Roxhed, Niclas; Linnros, Jan
2015-08-01
Solid state nanopores enable translocation and detection of single bio-molecules such as DNA in buffer solutions. Here, sub-10 nm nanopore arrays in silicon membranes were fabricated by using electron-beam lithography to define etch pits and by using a subsequent electrochemical etching step. This approach effectively decouples positioning of the pores and the control of their size, where the pore size essentially results from the anodizing current and time in the etching cell. Nanopores with diameters as small as 7 nm, fully penetrating 300 nm thick membranes, were obtained. The presented fabrication scheme to form large arrays of nanopores is attractive for parallel bio-molecule sensing and DNA sequencing using optical techniques. In particular the signal-to-noise ratio is improved compared to other alternatives such as nitride membranes suffering from a high-luminescence background.
Nanopore arrays in a silicon membrane for parallel single-molecule detection: fabrication.
Schmidt, Torsten; Zhang, Miao; Sychugov, Ilya; Roxhed, Niclas; Linnros, Jan
2015-08-07
Solid state nanopores enable translocation and detection of single bio-molecules such as DNA in buffer solutions. Here, sub-10 nm nanopore arrays in silicon membranes were fabricated by using electron-beam lithography to define etch pits and by using a subsequent electrochemical etching step. This approach effectively decouples positioning of the pores and the control of their size, where the pore size essentially results from the anodizing current and time in the etching cell. Nanopores with diameters as small as 7 nm, fully penetrating 300 nm thick membranes, were obtained. The presented fabrication scheme to form large arrays of nanopores is attractive for parallel bio-molecule sensing and DNA sequencing using optical techniques. In particular the signal-to-noise ratio is improved compared to other alternatives such as nitride membranes suffering from a high-luminescence background.
NASA Astrophysics Data System (ADS)
Laudien, Robert; Schultze, Rainer; Wieser, Jochen
2010-10-01
In this contribution two analytical devices for the fast detection of security-relevant substances like narcotics and explosives are presented. One system is based on an ion trap mass spectrometer (ITMS) with single photon ionization (SPI). This soft ionization technique, unlike electron impact ionization (EI), reduces unwanted fragment ions in the mass spectra allowing the clear determination of characteristic (usually molecular) ions. Their enrichment in the ion trap and identification by tandem MS investigations (MS/MS) enables the detection of the target substances in complex matrices at low concentrations without time-consuming sample preparation. For SPI an electron beam pumped excimer light source of own fabrication (E-Lux) is used. The SPI-ITMS system was characterized by the analytical study of different drugs like cannabis, heroin, cocaine, amphetamines, and some precursors. Additionally, it was successfully tested on-site in a closed illegal drug laboratory, where low quantities of MDMA could be directly detected in samples from floors, walls and lab equipments. The second analytical system is based on an ion mobility (IM) spectrometer with resonant multiphoton ionization (REMPI). With the frequency quadrupled Nd:YAG laser (266 nm), used for ionization, a selective and sensitive detection of aromatic compounds is possible. By application of suited aromatic dopants, in addition, also non-aromatic polar compounds are accessible by ion molecule reactions like proton transfer or complex formation. Selected drug precursors could be successfully detected with this device as well, qualifying it to a lower-priced alternative or useful supplement of the SPI-ITMS system for security analysis.
Jung, Seung-Ryoung; Han, Rui; Sun, Wei; Jiang, Yifei; Fujimoto, Bryant S; Yu, Jiangbo; Kuo, Chun-Ting; Rong, Yu; Zhou, Xing-Hua; Chiu, Daniel T
2018-05-15
We describe here a flow platform for quantifying the number of biomolecules on individual fluorescent nanoparticles. The platform combines line-confocal fluorescence detection with near nanoscale channels (1-2 μm in width and height) to achieve high single-molecule detection sensitivity and throughput. The number of biomolecules present on each nanoparticle was determined by deconvolving the fluorescence intensity distribution of single-nanoparticle-biomolecule complexes with the intensity distribution of single biomolecules. We demonstrate this approach by quantifying the number of streptavidins on individual semiconducting polymer dots (Pdots); streptavidin was rendered fluorescent using biotin-Alexa647. This flow platform has high-throughput (hundreds to thousands of nanoparticles detected per second) and requires minute amounts of sample (∼5 μL at a dilute concentration of 10 pM). This measurement method is an additional tool for characterizing synthetic or biological nanoparticles.
Inflammable Gas Mixture Detection with a Single Catalytic Sensor Based on the Electric Field Effect
Tong, Ziyuan; Tong, Min-Ming; Meng, Wen; Li, Meng
2014-01-01
This paper introduces a new way to analyze mixtures of inflammable gases with a single catalytic sensor. The analysis technology was based on a new finding that an electric field on the catalytic sensor can change the output sensitivity of the sensor. The analysis of mixed inflammable gases results from processing the output signals obtained by adjusting the electric field parameter of the catalytic sensor. For the signal process, we designed a group of equations based on the heat balance of catalytic sensor expressing the relationship between the output signals and the concentration of gases. With these equations and the outputs of different electric fields, the gas concentration in a mixture could be calculated. In experiments, a mixture of methane, butane and ethane was analyzed by this new method, and the results showed that the concentration of each gas in the mixture could be detected with a single catalytic sensor, and the maximum relative error was less than 5%. PMID:24717635
Slator, Paddy J.; Cairo, Christopher W.; Burroughs, Nigel J.
2015-01-01
We develop a Bayesian analysis framework to detect heterogeneity in the diffusive behaviour of single particle trajectories on cells, implementing model selection to classify trajectories as either consistent with Brownian motion or with a two-state (diffusion coefficient) switching model. The incorporation of localisation accuracy is essential, as otherwise false detection of switching within a trajectory was observed and diffusion coefficient estimates were inflated. Since our analysis is on a single trajectory basis, we are able to examine heterogeneity between trajectories in a quantitative manner. Applying our method to the lymphocyte function-associated antigen 1 (LFA-1) receptor tagged with latex beads (4 s trajectories at 1000 frames s−1), both intra- and inter-trajectory heterogeneity were detected; 12–26% of trajectories display clear switching between diffusive states dependent on condition, whilst the inter-trajectory variability is highly structured with the diffusion coefficients being related by D 1 = 0.68D 0 − 1.5 × 104 nm2 s−1, suggestive that on these time scales we are detecting switching due to a single process. Further, the inter-trajectory variability of the diffusion coefficient estimates (1.6 × 102 − 2.6 × 105 nm2 s−1) is very much larger than the measurement uncertainty within trajectories, suggesting that LFA-1 aggregation and cytoskeletal interactions are significantly affecting mobility, whilst the timescales of these processes are distinctly different giving rise to inter- and intra-trajectory variability. There is also an ‘immobile’ state (defined as D < 3.0 × 103 nm2 s−1) that is rarely involved in switching, immobility occurring with the highest frequency (47%) under T cell activation (phorbol-12-myristate-13-acetate (PMA) treatment) with enhanced cytoskeletal attachment (calpain inhibition). Such ‘immobile’ states frequently display slow linear drift, potentially reflecting binding to a dynamic actin cortex
Li, Huiyu; Zhao, Li; Xu, Yuan; Zhou, Tianyu; Liu, Haochi; Huang, Ning; Ding, Jie; Li, Yi; Ding, Lan
2018-08-01
It is difficult to detect tetracycline (TC) in honey sample by using carbon dots (CDs) because the autofluorescence of the matrix of honey sample overlaps with the fluorescence emission spectrum of the large majority of CDs. Herein, single-hole hollow molecularly imprinted polymers embedded carbon dots (HMIP@CD) was prepared via microwave-assisted method. TC in diluted honey sample was adsorbed by the HMIP@CD within 3 min, after which the HMIP@CD absorbed with TC was separated by centrifugation from honey sample and redispersed into phosphate buffer solution. The autofluorescence of honey that interferes with the fluorescence signal of HMIP@CD was avoided. The method exhibited an excellent linearity within 10-200 μg L -1 and a low detection limit of 3.1 μg L -1 . At three spiking levels of TC, the recoveries ranged from 93% to 105% with precisions below 1.6%. This method provides an effective strategy for detecting analyte in complex matrix with autofluorescence interference. Copyright © 2018. Published by Elsevier B.V.
Single Nanoparticle Plasmonic Sensors
Sriram, Manish; Zong, Kelly; Vivekchand, S. R. C.; Gooding, J. Justin
2015-01-01
The adoption of plasmonic nanomaterials in optical sensors, coupled with the advances in detection techniques, has opened the way for biosensing with single plasmonic particles. Single nanoparticle sensors offer the potential to analyse biochemical interactions at a single-molecule level, thereby allowing us to capture even more information than ensemble measurements. We introduce the concepts behind single nanoparticle sensing and how the localised surface plasmon resonances of these nanoparticles are dependent upon their materials, shape and size. Then we outline the different synthetic approaches, like citrate reduction, seed-mediated and seedless growth, that enable the synthesis of gold and silver nanospheres, nanorods, nanostars, nanoprisms and other nanostructures with tunable sizes. Further, we go into the aspects related to purification and functionalisation of nanoparticles, prior to the fabrication of sensing surfaces. Finally, the recent developments in single nanoparticle detection, spectroscopy and sensing applications are discussed. PMID:26473866
Single Color Multiplexed ddPCR Copy Number Measurements and Single Nucleotide Variant Genotyping.
Wood-Bouwens, Christina M; Ji, Hanlee P
2018-01-01
Droplet digital PCR (ddPCR) allows for accurate quantification of genetic events such as copy number variation and single nucleotide variants. Probe-based assays represent the current "gold-standard" for detection and quantification of these genetic events. Here, we introduce a cost-effective single color ddPCR assay that allows for single genome resolution quantification of copy number and single nucleotide variation.
Prins, Theo W; van Dijk, Jeroen P; Beenen, Henriek G; Van Hoef, AM Angeline; Voorhuijzen, Marleen M; Schoen, Cor D; Aarts, Henk JM; Kok, Esther J
2008-01-01
Background To maintain EU GMO regulations, producers of new GM crop varieties need to supply an event-specific method for the new variety. As a result methods are nowadays available for EU-authorised genetically modified organisms (GMOs), but only to a limited extent for EU-non-authorised GMOs (NAGs). In the last decade the diversity of genetically modified (GM) ingredients in food and feed has increased significantly. As a result of this increase GMO laboratories currently need to apply many different methods to establish to potential presence of NAGs in raw materials and complex derived products. Results In this paper we present an innovative method for detecting (approved) GMOs as well as the potential presence of NAGs in complex DNA samples containing different crop species. An optimised protocol has been developed for padlock probe ligation in combination with microarray detection (PPLMD) that can easily be scaled up. Linear padlock probes targeted against GMO-events, -elements and -species have been developed that can hybridise to their genomic target DNA and are visualised using microarray hybridisation. In a tenplex PPLMD experiment, different genomic targets in Roundup-Ready soya, MON1445 cotton and Bt176 maize were detected down to at least 1%. In single experiments, the targets were detected down to 0.1%, i.e. comparable to standard qPCR. Conclusion Compared to currently available methods this is a significant step forward towards multiplex detection in complex raw materials and derived products. It is shown that the PPLMD approach is suitable for large-scale detection of GMOs in real-life samples and provides the possibility to detect and/or identify NAGs that would otherwise remain undetected. PMID:19055784
Prins, Theo W; van Dijk, Jeroen P; Beenen, Henriek G; Van Hoef, Am Angeline; Voorhuijzen, Marleen M; Schoen, Cor D; Aarts, Henk J M; Kok, Esther J
2008-12-04
To maintain EU GMO regulations, producers of new GM crop varieties need to supply an event-specific method for the new variety. As a result methods are nowadays available for EU-authorised genetically modified organisms (GMOs), but only to a limited extent for EU-non-authorised GMOs (NAGs). In the last decade the diversity of genetically modified (GM) ingredients in food and feed has increased significantly. As a result of this increase GMO laboratories currently need to apply many different methods to establish to potential presence of NAGs in raw materials and complex derived products. In this paper we present an innovative method for detecting (approved) GMOs as well as the potential presence of NAGs in complex DNA samples containing different crop species. An optimised protocol has been developed for padlock probe ligation in combination with microarray detection (PPLMD) that can easily be scaled up. Linear padlock probes targeted against GMO-events, -elements and -species have been developed that can hybridise to their genomic target DNA and are visualised using microarray hybridisation.In a tenplex PPLMD experiment, different genomic targets in Roundup-Ready soya, MON1445 cotton and Bt176 maize were detected down to at least 1%. In single experiments, the targets were detected down to 0.1%, i.e. comparable to standard qPCR. Compared to currently available methods this is a significant step forward towards multiplex detection in complex raw materials and derived products. It is shown that the PPLMD approach is suitable for large-scale detection of GMOs in real-life samples and provides the possibility to detect and/or identify NAGs that would otherwise remain undetected.
Wang, Junxiu; Xiong, Guoliang; Ma, Liang; Wang, Shihui; Zhou, Xu; Wang, Lei; Xiao, Lehui; Su, Xin; Yu, Changyuan
2017-08-15
Single-nucleotide mutation (SNM) has proven to be associated with a variety of human diseases. Development of reliable methods for the detection of SNM is crucial for molecular diagnosis and personalized medicine. The sandwich assays are widely used tools for detecting nucleic acid biomarkers due to their low cost and rapid signaling. However, the poor hybridization specificity of signal probe at room temperature hampers the discrimination of mutant and wild type. Here, we demonstrate a dynamic sandwich assay on magnetic beads for SNM detection based on the transient binding between signal probe and target. By taking the advantage of mismatch sensitive thermodynamics of transient DNA binding, the dynamic sandwich assay exhibits high discrimination factor for mutant with a broad range of salt concentration at room temperature. The beads used in this assay serve as a tool for separation, and might be helpful to enhance SNM selectivity. Flexible design of signal probe and facile magnetic separation allow multiple-mode downstream analysis including colorimetric detection and isothermal amplification. With this method, BRAF mutations in the genomic DNA extracted from cancer cell lines were tested, allowing sensitive detection of SNM at very low abundances (0.1-0.5% mutant/wild type). Copyright © 2017 Elsevier B.V. All rights reserved.
Del Ben, Fabio; Turetta, Matteo; Celetti, Giorgia; Piruska, Aigars; Bulfoni, Michela; Cesselli, Daniela; Huck, Wilhelm T S; Scoles, Giacinto
2016-07-18
The number of circulating tumor cells (CTCs) in blood is strongly correlated with the progress of metastatic cancer. Current methods to detect CTCs are based on immunostaining or discrimination of physical properties. Herein, a label-free method is presented exploiting the abnormal metabolic behavior of cancer cells. A single-cell analysis technique is used to measure the secretion of acid from individual living tumor cells compartmentalized in microfluidically prepared, monodisperse, picoliter (pL) droplets. As few as 10 tumor cells can be detected in a background of 200 000 white blood cells and proof-of-concept data is shown on the detection of CTCs in the blood of metastatic patients. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
Quantum cryptography using single-particle entanglement
DOE Office of Scientific and Technical Information (OSTI.GOV)
Lee, Jae-Weon; Lee, Eok Kyun; Chung, Yong Wook
2003-07-01
A quantum cryptography scheme based on entanglement between a single-particle state and a vacuum state is proposed. The scheme utilizes linear optics devices to detect the superposition of the vacuum and single-particle states. Existence of an eavesdropper can be detected by using a variant of Bell's inequality.
Guo, Qingsheng; Bai, Zhixiong; Liu, Yuqian; Sun, Qingjiang
2016-03-15
In this work, we report the application of streptavidin-coated quantum dot (strAV-QD) in molecular beacon (MB) microarray assays by using the strAV-QD to label the immobilized MB, avoiding target labeling and meanwhile obviating the use of amplification. The MBs are stem-loop structured oligodeoxynucleotides, modified with a thiol and a biotin at two terminals of the stem. With the strAV-QD labeling an "opened" MB rather than a "closed" MB via streptavidin-biotin reaction, a sensitive and specific detection of label-free target DNA sequence is demonstrated by the MB microarray, with a signal-to-background ratio of 8. The immobilized MBs can be perfectly regenerated, allowing the reuse of the microarray. The MB microarray also is able to detect single nucleotide polymorphisms, exhibiting genotype-dependent fluorescence signals. It is demonstrated that the MB microarray can perform as a 4-to-2 encoder, compressing the genotype information into two outputs. Copyright © 2015 Elsevier B.V. All rights reserved.
Detecting single-electron events in TEM using low-cost electronics and a silicon strip sensor.
Gontard, Lionel C; Moldovan, Grigore; Carmona-Galán, Ricardo; Lin, Chao; Kirkland, Angus I
2014-04-01
There is great interest in developing novel position-sensitive direct detectors for transmission electron microscopy (TEM) that do not rely in the conversion of electrons into photons. Direct imaging improves contrast and efficiency and allows the operation of the microscope at lower energies and at lower doses without loss in resolution, which is especially important for studying soft materials and biological samples. We investigate the feasibility of employing a silicon strip detector as an imaging detector for TEM. This device, routinely used in high-energy particle physics, can detect small variations in electric current associated with the impact of a single charged particle. The main advantages of using this type of sensor for direct imaging in TEM are its intrinsic radiation hardness and large detection area. Here, we detail design, simulation, fabrication and tests in a TEM of the front-end electronics developed using low-cost discrete components and discuss the limitations and applications of this technology for TEM.
Huang, Jiyong; Luo, Xiliang; Lee, Innam; Hu, Yushi; Cui, Xinyan Tracy; Yun, Minhee
2011-01-01
Single polypyrrole (PPy) nanowire-based microfluidic aptasensors were fabricated using a one-step electrochemical deposition method. The successful incorporation of the aptamers into the PPy nanowire was confirmed by fluorescence microscopy image. The microfluidic aptasensor showed responses to IgE protein solutions in the range from 0.01 nM to 100 nM, and demonstrated excellent specificity and sensitivity with faster response and rapid stabilization times (~20 s). At the lowest examined IgE concentration of 0.01nM, the microfluidic aptasensor still exhibited ~0.32% change in the conductance. The functionality of this aptasensor was able to be regenerated using an acid treatment with no major change in sensitivity. In addition, the detection of cancer biomarker MUC1 was performed using another microfluidic aptasensor, which showed a very low detection limit of 2.66 nM MUC1 compared to commercially available MUC1 diagnosis assay (800 nM). PMID:21937215
NASA Astrophysics Data System (ADS)
Rajesh, Sharma, Vikash; Puri, Nitin K.; Mulchandani, Ashok; Kotnala, Ravinder K.
2016-12-01
We report a single-walled carbon nanotube (SWNT) field-effect transistor (FET) functionalized with Polyamidoamine (PAMAM) dendrimer with 128 carboxyl groups as anchors for site specific biomolecular immobilization of protein antibody for C-reactive protein (CRP) detection. The FET device was characterized by scanning electron microscopy and current-gate voltage (I-Vg) characteristic studies. A concentration-dependent decrease in the source-drain current was observed in the regime of clinical significance, with a detection limit of ˜85 pM and a high sensitivity of 20% change in current (ΔI/I) per decade CRP concentration, showing SWNT being locally gated by the binding of CRP to antibody (anti-CRP) on the FET device. The low value of the dissociation constant (Kd = 0.31 ± 0.13 μg ml-1) indicated a high affinity of the device towards CRP analyte arising due to high anti-CRP loading with a better probe orientation on the 3-dimensional PAMAM structure.
NASA Astrophysics Data System (ADS)
Zhou, Anran; Xie, Weixin; Pei, Jihong; Chen, Yapei
2018-02-01
For ship targets detection in cluttered infrared image sequences, a robust detection method, based on the probabilistic single Gaussian model of sea background in Fourier domain, is put forward. The amplitude spectrum sequences at each frequency point of the pure seawater images in Fourier domain, being more stable than the gray value sequences of each background pixel in the spatial domain, are regarded as a Gaussian model. Next, a probability weighted matrix is built based on the stability of the pure seawater's total energy spectrum in the row direction, to make the Gaussian model more accurate. Then, the foreground frequency points are separated from the background frequency points by the model. Finally, the false-alarm points are removed utilizing ships' shape features. The performance of the proposed method is tested by visual and quantitative comparisons with others.
NASA Astrophysics Data System (ADS)
Garzelli, Andrea; Zoppetti, Claudia; Pinelli, Gianpaolo
2017-10-01
Coastline detection in synthetic aperture radar (SAR) images is crucial in many application fields, from coastal erosion monitoring to navigation, from damage assessment to security planning for port facilities. The backscattering difference between land and sea is not always documented in SAR imagery, due to the severe speckle noise, especially in 1-look data with high spatial resolution, high sea state, or complex coastal environments. This paper presents an unsupervised, computationally efficient solution to extract the coastline acquired by only one single-polarization 1-look SAR image. Extensive tests on Spotlight COSMO-SkyMed images of complex coastal environments and objective assessment demonstrate the validity of the proposed procedure which is compared to state-of-the-art methods through visual results and with an objective evaluation of the distance between the detected and the true coastline provided by regional authorities.
POTENTIAL FOR REDUCING STYRENE EXPOSURES FROM COPIED PAPER THROUGH USE OF LOW-EMITTING TONERS
The paper reports results of tests, conducted using 53-L chambers to determine styrene emission rates from freshly copied paper produced on a single photocopier using two toners manufactured for the copier having different styrene contents. Copied-paper styrene emissions with bot...
Yen, Yi-Kuang; Lai, Yu-Cheng; Hong, Wei-Ting; Pheanpanitporn, Yotsapoom; Chen, Chuin-Shan; Huang, Long-Sun
2013-01-01
This study demonstrates a novel method for electrical detection of C-reactive protein (CRP) as a means of identifying an infection in the body, or as a cardiovascular disease risk assay. The method uses a single free-standing, thermally controlled piezoresistive microcantilever biosensor. In a commonly used sensing arrangement of conventional dual cantilevers in the Wheatstone bridge cir