Three-color immunofluorescence imaging of Drosophila embryos by laser scanning confocal microscopy

Biotechniques. 1993 Jan;14(1):42-8.


We present a simple means for triple-labeling biological specimens by immunofluorescence using a laser scanning confocal microscope for imaging with a krypton/argon laser as a light source. Three separate images of fluorescein-, lissamine rhodamine- and cyanine-5-labeled antibodies are collected and subsequently merged to form the triple-labeled image, which is displayed at full-image resolution (24 bit) on a second image processing system. The technique is illustrated using immunofluorescence localization of three segmentation proteins in Drosophila embryos.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biotechnology
  • Color
  • Computer Graphics
  • Drosophila melanogaster
  • Embryo, Nonmammalian / anatomy & histology
  • Embryo, Nonmammalian / metabolism
  • Fluorescent Antibody Technique*
  • Image Processing, Computer-Assisted
  • Lasers
  • Microscopy, Fluorescence / methods*
  • Proteins / metabolism


  • Proteins