Simultaneous two-photon excitation of distinct labels for dual-color fluorescence crosscorrelation analysis

Confocal fluorescence correlation spectroscopy as a time-averaging fluctuation analysis combining maximum sensitivity with high statistical confidence has proved to be a very versatile and powerful tool for detection and temporal investigation of biomolecules at ultralow concentrations on surfaces, in solutions, and in living cells. To probe the interaction of different molecular species for a detailed understanding of biologically relevant mechanisms, crosscorrelation studies on dual or multiple fluorophore assays with spectrally distinct excitation and emission are particularly promising. Despite the considerable improvement of detection specificity provided by fluorescence crosscorrelation analysis, few applications have so far been reported, presumably because of the practical challenges of properly aligning and controlling the stability of the experimental setup. In this work, we demonstrate that two-photon excitation combined with dual-color fluorescence correlation spectroscopy can be the key to simplifying simultaneous investigations of multiple fluorescent species significantly on a single-molecule scale. Two-photon excitation allows accession of common fluorophores of largely distinct emission by the same excitation wavelength, because differences in selection rules and vibronic coupling can induce considerable shifts between the one-photon and two-photon excitation spectra. The concept of dual-color two-photon fluorescence crosscorrelation analysis is introduced and experimentally demonstrated with an established assay probing the selective cleavage of dual-labeled DNA substrates by restriction endonuclease EcoRI.

The effect of pre-clinical fieldwork subject on the confidence and professional skills of undergraduate occupational therapy students

Objectives: This study investigated the effect of a pre-clinical fieldwork subject on the confidence and professional skills of undergraduate occupational therapy students during their first full-time clinical fieldwork placement. Methods: Participants were 31 third year students enrolled in the pre-clinical fieldwork subject (experimental group), and 25 students not enrolled in the subject (control group). Both quantitative and qualitative methodologies were employed to address the research question. Student anxiety levels were measured using the State-Trait Anxiety Inventory (STAI) and compared at three different intervals - prior to commencing the university semester, prior to commencing full-time practical placement, and following this full-time placement. Individual interviews were conducted with six students (three from the experimental group, three from the control group) to explore their perceptions regarding confidence levels and skill proficiencies during full-time placement. At each data analysis interval, responses for both groups were compared using independent samples t-tests. Responses were compared over time using analysis of variance (ANOVA). The qualitative data were subjected to thematic and content analysis. Results and Conclusions: In general, student anxiety levels did not differ between the experimental and control groups over time. However, qualitative results suggest that students who were enrolled in the pre-clinical subject have greater confidence and competence with occupational therapy skills, and have heightened awareness of the expectations on full-time placement.

Conditional Confidence Interval for the Scale Parameter of a Weibull Distribution

2000 Mathematics Subject Classification: 62F25, 62F03.

Effect of delay of first interview and repetition of interview on accuracy and confidence of recall of a flashbulb -type memory

Hearing of the news of the death of Diana, Princess of Wales, in a traffic accident, is taken as an analogue for being a percipient but uninvolved witness to a crime, or a witness to another person's sudden confession to some illegal act. This event (known in the literature as a “reception event”) has previously been hypothesized to cause one to form a special type of memory commonly known as a “flashbulb memory” (FB) (Brown and Kulik, 1977). FB's are hypothesized to be especially resilient against forgetting, highly detailed including peripheral details, clear, and inspiring great confidence in the individual for their accuracy. FB's are dependent for their formation upon surprise, emotional valence, and impact, or consequentiality to the witness of the initiating event. FB's are thought to be enhanced by frequent rehearsal. FB's are very important in the context of criminal investigation and litigation in that investigators and jurors usually place great store in witnesses, regardless of their actual accuracy, who claim to have a clear and complete recollection of an event, and who express this confidently. Therefore, the lives, or at least the freedom, of criminal defendants, and the fortunes of civil litigants hang on the testimony of witnesses professing to have FB's. ^ In this study, which includes a large and diverse sample (N = 305), participants were surveyed within 2–4 days after hearing of the fatal accident, and again at intervals of 2 and 4 weeks, 6, 12, and 18 months. Contrary to the FB hypothesis, I found that participants' FB's degraded over time beginning at least as early as two weeks post event. At about 12 months the memory trace stabilized, resisting further degradation. Repeated interviewing did not have any negative affect upon accuracy, contrary to concerns in the literature. Analysis by correlation and regression indicated no effect or predictive power for participant age, emotionality, confidence, or student status, as related to accuracy of recall; nor was participant confidence in accuracy predicted by emotional impact as hypothesized. Results also indicate that, contrary to the notions of investigators and jurors, witnesses become more inaccurate over time regardless of their confidence in their memories, even for highly emotional events. ^

A theoretical analysis of eyewitness identification: Dual -process theory, signal detection theory and eyewitness confidence

Given the growing number of wrongful convictions involving faulty eyewitness evidence and the strong reliance by jurors on eyewitness testimony, researchers have sought to develop safeguards to decrease erroneous identifications. While decades of eyewitness research have led to numerous recommendations for the collection of eyewitness evidence, less is known regarding the psychological processes that govern identification responses. The purpose of the current research was to expand the theoretical knowledge of eyewitness identification decisions by exploring two separate memory theories: signal detection theory and dual-process theory. This was accomplished by examining both system and estimator variables in the context of a novel lineup recognition paradigm. Both theories were also examined in conjunction with confidence to determine whether it might add significantly to the understanding of eyewitness memory. ^ In two separate experiments, both an encoding and a retrieval-based manipulation were chosen to examine the application of theory to eyewitness identification decisions. Dual-process estimates were measured through the use of remember-know judgments (Gardiner & Richardson-Klavehn, 2000). In Experiment 1, the effects of divided attention and lineup presentation format (simultaneous vs. sequential) were examined. In Experiment 2, perceptual distance and lineup response deadline were examined. Overall, the results indicated that discrimination and remember judgments (recollection) were generally affected by variations in encoding quality and response criterion and know judgments (familiarity) were generally affected by variations in retrieval options. Specifically, as encoding quality improved, discrimination ability and judgments of recollection increased; and as the retrieval task became more difficult there was a shift toward lenient choosing and more reliance on familiarity. ^ The application of signal detection theory and dual-process theory in the current experiments produced predictable results on both system and estimator variables. These theories were also compared to measures of general confidence, calibration, and diagnosticity. The application of the additional confidence measures in conjunction with signal detection theory and dual-process theory gave a more in-depth explanation than either theory alone. Therefore, the general conclusion is that eyewitness identifications can be understood in a more complete manor by applying theory and examining confidence. Future directions and policy implications are discussed. ^

Lineup identification accuracy: the effects of alcohol, target presence, confidence ratings, and response time

Despite the intoxication of many eyewitnesses at crime scenes, only four published studies to date have investigated the effects of alcohol intoxication on eyewitness identification performance. While one found intoxication significantly increased false identification rates from target absent showups, three found no such effect using the more traditional lineup procedure. The present study sought to further explore the effects of alcohol intoxication on identification performance and examine whether accurate decisions from intoxicated witnesses could be postdicted by confidence and response times. One hundred and twenty participants engaged in a study examining the effects of intoxication (control, placebo, and mild intoxication) and target presence on identification performance. Participants viewed a simultaneous lineup one week after watching a mock crime video of a man attempting to steal cars. Ethanol intoxication (0.6 ml/kg) was found to make no significant difference to identification accuracy and such identifications from intoxicated individuals were made no less confidently or slowly than those from sober witnesses. These results are discussed with respect to the previous research examining intoxicated witness identification accuracy and the misconceptions the criminal justice system holds about the accuracy of such witnesses.