Quality control in single-molecule studies of kinesins and microtubule-associated proteins.
Commercial microscopes capable of single-molecule experiments have made it simple for researchers to adopt these powerful techniques. This chapter is meant to help newcomers assess whether their data is of sufficient quality to warrant time-intensive analysis. Two problems can hamper single-molecule experiments: (1) non-specific aggregation of the proteins of interest and (2) detection thresholds from a poor microscope setup. I outline four steps that researchers can take to overcome these problems and convince themselves that they are observing bona fide single molecules.
Brouhard
Gary J
GJ
Department of Biology, McGill University, Montréal, Québec H3A1B1, Canada.
D000818:Animals; D019411:Clinical Laboratory Techniques; D005455:Fluorescent Antibody Technique / Q000379:methods; D006801:Humans; D016547:Kinesins / Q000032:analysis* / Q000378:metabolism; D008856:Microscopy, Fluorescence / Q000379:methods; D008869:Microtubule-Associated Proteins / Q000032:analysis* / Q000378:metabolism; D038761:Photobleaching; D011786:Quality Control
D008869:Microtubule-Associated Proteins; D016547:Kinesins