Fluorescence photobleaching analysis for the study of cellular dynamics.
The wide availability of the confocal microscope and the emergence of green fluorescent protein (GFP) transfection technology has led to the increasing use of photobleaching studies to examine aspects of cellular dynamics in living cells. In this review, we examine the theory and practice of performing photobleaching studies using a confocal microscope. We illustrate the application of photobleaching protocols using our own measurements of fluorescently labelled red blood cells and of malaria parasite-infected erythrocytes expressing GFP fusions and examine other examples from the literature.
Klonis
Nectarios
N
Department of Biochemistry, La Trobe University, Bundoora, Victoria, Australia.
Rug
Melanie
M
Harper
Ian
I
Wickham
Mark
M
Cowman
Alan
A
Tilley
Leann
L
D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D002451:Cell Compartmentation; D004912:Erythrocytes / Q000378:metabolism / Q000469:parasitology; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D008288:Malaria / Q000097:blood / Q000469:parasitology; D018613:Microscopy, Confocal; D008954:Models, Biological; D038761:Photobleaching; D011993:Recombinant Fusion Proteins / Q000378:metabolism
D005456:Fluorescent Dyes; D008164:Luminescent Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins