Visualization of identified GFP-expressing cells by light and electron microscopy. J Histochem Cytochem 2003 Mar;51(3):271-4
Date
02/18/2003Pubmed ID
12588954DOI
10.1177/002215540305100301Scopus ID
2-s2.0-0037369967 (requires institutional sign-in at Scopus site) 67 CitationsAbstract
We have developed a procedure for visualizing GFP expression in fixed tissue after embedding in LR White. We find that GFP fluorescence survives fixation in 4% paraformaldehyde/0.1% glutaraldehyde and can be visualized directly by fluorescence microscopy in unstained, 1 microm sections of LR White-embedded material. The antigenicity of the GFP is retained in these preparations, so that GFP localization can be visualized in the electron microscope after immunogold labeling with anti-GFP antibodies. The ultrastructural morphology of tissue fixed and embedded by this protocol is of quality sufficient for subcellular localization of GFP. Thus, expression of GFP constructs can be visualized in living tissue and the same cells relocated in semithin sections. Furthermore, semithin sections can be used to locate GFP-expressing cells for examination by immunoelectron microscopy of the same material after thin sectioning.
Author List
Luby-Phelps K, Ning G, Fogerty J, Besharse JCAuthor
Joseph C. Besharse PhD, MA Emeritus Professor in the Cell Biology Neurobiology and Anatomy department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Acrylic ResinsAnimals
Eye
Green Fluorescent Proteins
Luminescent Proteins
Microscopy, Fluorescence
Microscopy, Immunoelectron
Tissue Embedding
Tissue Fixation
Zebrafish
