Three-color immunofluorescence imaging of Drosophila embryos by laser scanning confocal microscopy
Paddock, S.W.; Langeland, J.A.; DeVries, P.J.; Carroll, S.B.
Biotechniques 14(1): 42-48
1993
ISSN/ISBN: 0736-6205 PMID: 8424875 Document Number: 407219
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.