Laser Scanning Cytometry for selection of green fluorescent protein transgenic mice using small number of blood cells.
A Laser Scanning Cytometry-based method was developed for identification of transgenic mice expressing green fluorescent protein (GFP) using minute amounts of peripheral blood. The difference between the autofluorescence of cells not expressing GFP and the fluorescence of GFP expressing cells after excitation with Ar-ion laser (wavelength 488 nm) and detection of emitted fluorescent light in the green channel was high enough for unambiguous identification of the GFP expressing mice. The sensitivity of this method was estimated 1:10(4) for detection of rare GFP expressing cells under the conditions used. This sensitivity should be sufficient for many studies on microchimerism. Because of the possibility for relocation of the cells, this method will be particularly useful for characterizing the cells with high GFP expression using other markers of cell phenotype or conventional morphological analysis.
Megyeri
Attila
A
Barbara Ann Karmanos Cancer Institute, Wayne State University School of Medicine, 4100 John R, Detroit, MI 48201, USA. amegy@jaguar.dote.hu
Kaplan
Joseph
J
Eliason
James F
JF
D000818:Animals; D002478:Cells, Cultured; D005260:Female; D020869:Gene Expression Profiling / Q000379:methods*; D005819:Genetic Markers; D005820:Genetic Testing / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000097:blood* / Q000235:genetics*; D047569:Laser Scanning Cytometry / Q000379:methods*; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic / Q000378:metabolism*; D011994:Recombinant Proteins / Q000032:analysis / Q000378:metabolism
D005819:Genetic Markers; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins