Visualization of the microtubules of glutaraldehyde-fixed cells by reflection-enhanced backscatter confocal microscopy.
Performing reflection-mode (backscatter-mode) confocal microscopy on cells growing on reflective substrates gives images that have improved contrast and are more easily interpreted than standard reflection-mode confocal micrographs (Keith et al., 1998). However, a number of factors degrade the quality of images taken with the highest-resolution microscope objectives in this technique. We here describe modifications to reflection-enhanced backscatter confocal microscopy that (partially) overcome these factors. With these modifications of the technique, it is possible to visualize structures the size-and refractility-of individual microtubules in intact cells. Additionally, we demonstrate that this technique, in common with fluorescence techniques such as standing wave widefield fluorescence microscopy and 4-Pi confocal microscopy, offers improved resolution in the Z-direction.
Keith
Charles H
CH
Department of Cellular Biology, The University of Georgia, 724 Biological Sciences Building, Athens, GA 30602, USA.
Farmer
Mark A
MA
D000818:Animals; D002460:Cell Line; D007668:Kidney; D018613:Microscopy, Confocal / Q000379:methods*; D008870:Microtubules / Q000502:physiology / Q000648:ultrastructure*; D012680:Sensitivity and Specificity; D013692:Telophase