Confocal Microscopy

Confocal microscopy is used for imaging fluorescent signals, usually in fixed specimens. It generates high resolution optical sections.

This is a C. elegans embryo at the 2-cell stage (bigger AB cell at bottom, P1 at the top). Red represents the membrane, green represents the microtubules (MTs) and blue shows its chromosome. This image combines images from 3 different channels. The membrane, MTs and chromosome were imaged using 3 channels, color-mapped to 3 colors (red, green, and blue).

LSCM Compared to Widefield Fluorescence Microscopy

By applying pinhole aperature to LSCM:

(Eliminate out-of-focus flare, 90 KB, movie length: 52")

(The thickness of optical section is controllable, 160 KB, movie length: 15")

Credits

  1. LSCM image: Maria Vidal, the John White Lab at UW-Madison
  2. Images in "Eliminate out-of-flare" flash movie: Haining Zhang, the John White Lab at UW-Madison
  3. Images & movies in "The thickness of section is controllable" flash movie: Jayne Squirrell, the John White Lab at UW-Madison
PreviewAttachmentSize
ConfocalI.swf91.24 KB
ConfocalPinholeII.swf162.99 KB