Stochastic Blinking
Stochastic Blinking - Related Videos
08:52
The Attentional Blink
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2023
Source: Laboratory of Jonathan Flombaum—Johns Hopkins University In order for recognition of a certain stimulus to take place, visual attention needs to be directed towards said stimulus. To the earliest parts of the visual system, objects are not objects, they are collections of visual features-lines, corners, changes in texture, color, and light. Attention is the resource that is necessary for later processing in order to recognize what a given bundle of features adds up to. This makes...
08:45
A Dual Task Procedure Combined with Rapid Serial Visual Presentation to Test Attentional Blink for Nontargets
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2014
This paper describes a novel dual task procedure combined with Rapid Serial Visual Presentation to look at attentional blink at varied stimulus onset asynchronies. This experimental procedure differed from others in that it tested attentional blink for nontargets.
09:36
Direct Stochastic Optical Reconstruction Microscopy of Extracellular Vesicles in Three Dimensions
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Cited by 8 •
2021
Direct stochastic optical reconstruction microscopy (dSTORM) is used to bypass the typical diffraction limit of light microscopy and to view exosomes at the nanometer scale. It can be employed in both two and three dimensions to characterize exosomes.
05:52
Observation and Analysis of Blinking Surface-enhanced Raman Scattering
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Cited by 4 •
2018
This protocol describes the analysis of blinking surface-enhanced Raman scattering due to the random walk of a single molecule on a silver surface using power laws.
14:23
Imaging Intermediate Filaments and Microtubules with 2-dimensional Direct Stochastic Optical Reconstruction Microscopy
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Cited by 6 •
2018
The overall goal of this methodology is to give the optimal experimental conditions from sample preparation to image acquisition and reconstruction in order to perform 2D dual color dSTORM images of microtubules and intermediate filaments in fixed cells...
FAQs
Fluorophores switch probabilistically between emissive states that produce detectable light and nonemissive states that do not. These transitions can arise from reversible chemical reactions, formation of a triplet state, or temporary adoption of a dark molecular conformation. Because switching occurs over time, the same molecule may contribute detectable signal in some image frames but not others.
Several processes can generate the intermittent signal. Reversible chemical reactions can alter whether a fluorophore emits, triplet-state formation can temporarily interrupt fluorescence, and conformational changes can place the molecule in a dark state. Distinguishing these possible routes helps researchers interpret fluctuations as molecular behavior rather than treating every intensity change as equivalent.
Blinking separates molecules in time even when their signals overlap spatially. During one frame, only a subset of fluorophores may occupy detectable states, allowing their positions to be localized more distinctly. Repeating this observation across successive frames supplies the temporal information needed to separate overlapping emitters and reconstruct a higher-resolution image.
A typical workflow records fluorescence across successive frames, identifies molecules that are detectable in individual frames, and uses their localized positions to build a reconstruction. Molecules that overlap in the original image can appear separately when they blink at different times. The resulting reconstruction uses the accumulated single-molecule localizations to reveal spatial organization at higher resolution.
Bioengineers can analyze blinking to investigate how biomolecules are arranged within a sample. Localizations collected over time provide spatial information that can reveal organization not readily distinguished when many fluorophores emit simultaneously. This application connects the optical behavior of engineered fluorescent probes with questions about molecular distribution and the structure of biological systems.
Fluctuations provide information that can be analyzed alongside the locations of labeled molecules. In bioengineering, this supports quantitative studies of biomolecular interactions and the evaluation of engineered fluorescent probes. Researchers can therefore use blinking not only to form super-resolution images, but also to examine how probe behavior and molecular organization contribute to measured fluorescence patterns.