Every laboratory with a fluorescence microscope should consider counting molecules.
Protein numbers in cells determine rates of biological processes, influence the architecture of cellular structures, reveal the stoichiometries of protein complexes, guide in vitro biochemical reconstitutions, and provide parameter values for mathematical modeling. The purpose of this essay is to increase awareness of methods for counting protein molecules using fluorescence microscopy and encourage more cell biologists to report these numbers. We address the state of the field in terms of utility and accuracy of the numbers reported and point readers to references for details of specific techniques and applications.
Department of Molecular Genetics, The Ohio State University, Columbus, OH 43210 coffman.147@osu.edu wu.620@osu.edu.
Valerie C
VC
Coffman
Department of Molecular Genetics, The Ohio State University, Columbus, OH 43210Department of Molecular and Cellular Biochemistry, The Ohio State University, Columbus, OH 43210 coffman.147@osu.edu wu.620@osu.edu.
Jian-Qiu
JQ
Wu
D001324:Autoantigens / Q000032:analysis*; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone / Q000032:analysis*; D003598:Cytoskeletal Proteins / Q000032:analysis*; D004268:DNA-Binding Proteins / Q000032:analysis*; D005456:Fluorescent Dyes; D008856:Microscopy, Fluorescence / Q000379:methods*; D038761:Photobleaching; D012015:Reference Standards; D012016:Reference Values; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000032:analysis*; D012568:Schizosaccharomyces; D029702:Schizosaccharomyces pombe Proteins / Q000032:analysis*
D001324:Autoantigens; C092746:CSE4 protein, S cerevisiae; C531994:Cdc12 protein, S pombe; D000076247:Centromere Protein A; D002868:Chromosomal Proteins, Non-Histone; C410349:Cnp1 protein, S pombe; D003598:Cytoskeletal Proteins; D004268:DNA-Binding Proteins; D005456:Fluorescent Dyes; D029701:Saccharomyces cerevisiae Proteins; D029702:Schizosaccharomyces pombe Proteins