Three-color immunofluorescence imaging of Drosophila embryos by laser scanning confocal microscopy.
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.
Paddock
S W
SW
Howard Hughes Medical Institute, Laboratory of Molecular Biology, Madison, WI 53706.
Langeland
J A
JA
DeVries
P J
PJ
Carroll
S B
SB
D000818:Animals; D001709:Biotechnology; D003116:Color; D003196:Computer Graphics; D004331:Drosophila melanogaster; D004625:Embryo, Nonmammalian / Q000033:anatomy & histology / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D007091:Image Processing, Computer-Assisted; D007834:Lasers; D008856:Microscopy, Fluorescence / Q000379:methods*; D011506:Proteins / Q000378:metabolism
D011506:Proteins