Visualization of the microtubules of glutaraldehyde-fixed cells by reflection-enhanced backscatter confocal microscopy

Microsc Microanal. 2006 Apr;12(2):113-23. doi: 10.1017/S1431927606060016.

Abstract

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.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • Cell Line
  • Kidney
  • Microscopy, Confocal / methods*
  • Microtubules / physiology
  • Microtubules / ultrastructure*
  • Sensitivity and Specificity
  • Telophase