Similarly, if the fluorescent proteins bind to static cell receptors, the rate of recovery will be retarded by a factor related to the association and disassociation coefficients of binding. (wikipedia.org)
After the protein of interest is made fluorescent, generally by expression as a GFP fusion protein, a confocal microscope is used to photobleach and monitor a region of the cytoplasm, mitotic spindle, nucleus, or another cellular structure. (wikipedia.org)
The analysis is most simple when the fluorescence recovery is limited by either the rate of diffusion into the bleached area or by rate at which bleached proteins unbind from their binding sites within the bleached area, and are replaced by fluorescent protein. (wikipedia.org)
Single molecule imaging represents a subset of fluorescence microscopy techniques that uses fluorescent tags to detect and analyze individual single molecules. (photometrics.com)
Molecular1
Molecular dynamics can be measured using methods like fluorescence recovery after photo bleaching (FRET) or förster/fluorescence resonance energy transfer (FRET). (bsimaging.at)
Proteins1
Often the only dynamics considered are diffusion and binding/unbinding interactions, however, in principle proteins can also move via flow, i.e., undergo directed motion, and this was recognized very early by Axelrod et al. (wikipedia.org)
Quickly2
The fluorophores in this region receive high intensity illumination which causes their fluorescence lifetime to quickly elapse (limited to roughly 105 photons before extinction). (wikipedia.org)
Generally speaking, fluorophores for single molecule imaging should be: Bright (high extinction coefficient and high quantum yield) to maximize signal, photostable (do not photobleach quickly) to allow the fluorophore to be imaged for long periods, small enough to not disrupt the biological activity of the labeled molecule and emit light in the visible region of the spectrum, preferably where the scientific camera has the highest quantum efficiency. (photometrics.com)
At 4% and 8% isoflurane this structure is lost and replaced with randomized dynamics, as quantified by the percentage of total ensemble variance captured by the first three principal components. (asahq.org)
High3
Fluorescence microscopy is a workhorse technique in biological sciences allowing specific biological structures to be labeled and imaged with high contrast. (photometrics.com)
Unanesthetized animals display distinct global neuronal states that are reflected in a high degree of correlation ( R = 0.196 ± 0.070) between neurons and low-frequency, large-amplitude neuronal dynamics. (asahq.org)
0.0001 vs. unanesthetized) with high-frequency dynamics that are even further restricted. (asahq.org)
In order to evaluate the accessibility of the pretreated poplar samples, two fluorescent probes (rhodamine B-isothiocyanate-dextrans of 20 and 70 kDa) were selected, and their mobility was measured by using the fluorescence recovery after photobleaching (FRAP) technique in a full factorial experiment. (biomedcentral.com)
Cell1
Similarly, if the fluorescent proteins bind to static cell receptors, the rate of recovery will be retarded by a factor related to the association and disassociation coefficients of binding. (wikipedia.org)