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- W2043691452 abstract "Single particle tracking (SPT) has become a common technique for studying protein and lipid behavior in cell membranes. Using photo-stable fluorescent probes such as quantum dots, SPT is now being applied using multi-color strategies in order to look at nano-scale protein-protein interactions. However, the sparse labeling of each spectral species needed to limit the occurrence of fluorophores overlapping within the diffraction limit reduces the probability of overlap from opposing spectral species and the frequency of observed interactions. We present a single particle tracking method that allows a higher density of labels by performing a Bayesian multi-fluorophore fit to particles that are spaced closer than the diffraction limit. We call a single event of overlapping trajectories a ‘vertex.’ We use knowledge of the particles' intensities, diffusion constants positions before and after the vertex, and possibly blinking rates to improve the multi-fluorophore fits through the vertex. We examine the accuracy of the trajectories and fits as a function of label density. Furthermore, we show that under a range of frame and blinking rates, fluorescence intermittency from blinking probes can improve the accuracy of multi-fluorophore fits. We show that single particle and Bayesian multi-particle fits can be performed at more than 105 fits per second using an iterative fitting method implemented on GPU architecture." @default.
- W2043691452 created "2016-06-24" @default.
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- W2043691452 date "2010-01-01" @default.
- W2043691452 modified "2023-10-18" @default.
- W2043691452 title "High Density Single Particle Tracking Using Bayesian Multi-Fluorophore Fitting through Vertices" @default.
- W2043691452 doi "https://doi.org/10.1016/j.bpj.2009.12.3201" @default.
- W2043691452 hasPublicationYear "2010" @default.
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