Laser capture microdissection for gene expression analysis of specific cell populations in single blastocysts.
Laser capture microdissection (LCM) allows for the isolation of small tissue fractions from heterogeneous tissue sections, for downstream genetic or proteomic analysis without contamination by the surrounding tissue. This technique can also be successfully used for the isolation of small tissue fractions from developing embryos, such as expanding blastocysts. However, the small size of early-stage embryos hampers tissue processing prior to LCM. The present protocol describes the application of LCM to isolate specific cell fractions from blastocysts for downstream gene expression analysis with RT-PCR.
Department of Morphology, Faculty of Veterinary Medicine, Ghent University, Merelbeke, Belgium. Ward.DeSpiegelaere@UGent.be
Ward
W
De Spiegelaere
Muriel
M
Filliers
Ann
A
Van Soom
D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism; D002417:Cattle; D004867:Equipment Design; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D061346:Laser Capture Microdissection / Q000295:instrumentation / Q000379:methods*; D012313:RNA / Q000235:genetics / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D016707:Tissue Fixation / Q000379:methods
D012313:RNA