GFP detect in tissue section

"近藤 英介" kondo at CBSIMM01.M.CHIBA-U.AC.JP
Tue Aug 24 04:50:43 EST 1999


Iam now making knock GFP in some lymphocyte specific gene to report the gene
expression by GFP fluoresces.
In these mouse ,I can detect GAP fluoresces in thymocytes and splenocytes by
flowcytometry analysis(Very strong intensity).But I can not detect in frozen
fixed and
4%PFA fixed tissue section.
So I tried several method to detect.
Result 1 are law tissue can detected GFP fluoresces such hole organ ,a few
mm block and
single cell by fluoromicroscopy.3D scanning of comfocal laser microscopy is
good tool
for GFP positive cell detection.
Result 2 thick tissue cryo section(20um)can detected but not so clear cell
shape of GFP
positive cells  and dry up diminished fluoresces intensity.Under 20 um
section can not
detect GFP cells no longer.

Please advice to me  good  methods to detect GFP positive cells in tissue
section.


Eisuke Kondo ,Chiba Univ. ,Immunology



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