The Waterloo Institute for Nanotechnology (WIN) and the Department of Physics and Astronomy are pleased to present a seminar talk by Dr. Joshua N. Milstein, from the Department of Physics at the University of Toronto and the Department of Chemical and Physical Sciences, University of Toronto Mississauga.
Counting Single Molecules at the Nanoscale
Cell biology is becoming increasingly quantitative with advances in light microscopy strongly driving this trend. Beyond imaging structure, significant effort has gone into developing microscopy based approaches to determining the abundance of proteins and nucleic acids in cells. Molecular counting experiments can yield additional insight into cellular structure and define the stoichiometry of interacting protein complexes.
Moreover, since microscopy provides information at the single-cell level, it may be used to study stochastic variation within a population due to varying levels of mRNA and protein copy number, which is inaccessible to bulk techniques. Single-molecules localization microscopy (SMLM) has the potential to serve as an accurate, single-cell technique for counting the abundance of intracellular molecules.
However, the stochastic blinking of single fluorophores can introduce large uncertainties into the final count. Here we provide an foundation for applying SMLM to the problem of molecular counting based on the distribution of blinking events from a single fluorophore.
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