Unexpected photobleaching of Alexa 488 in a fixed bacterial sample during 2-photon excitation.
A sample of fixed bacterial cells was examined by immunofluorescence microscopy using an Alexa 488 conjugated secondary antibody for visualization. Excitation using visible light confirmed the expected photostability of this fluorophore; however, when using 2-photon excitation, Alexa 488 was rapidly and substantially photobleached. The unexpected instability of Alexa 488 under certain conditions may have deleterious consequences if not anticipated and accommodated in experimental protocols.
Peters
P C
PC
Institute for the Biotechnology of Infectious Diseases, University of Technology, Sydney, 2007, Australia. phoebe.c.peters@student.uts.edu.au
Thoni
C
C
Harry
E J
EJ
D001412:Bacillus subtilis; D006834:Hydrazines; D008856:Microscopy, Fluorescence; D038761:Photobleaching; D013194:Staining and Labeling
C417561:Alexa 488 hydrazide; D006834:Hydrazines