Using DSP, a reversible cross-linker, to fix tissue sections for immunostaining, microdissection and expression profiling.
Mammalian organs are typically comprised of several cell populations. Some (e.g. brain) are very heterogeneous, and this cellular complexity makes it difficult, if not impossible, to interpret expression profiles obtained with microarrays. Instruments, such as those manufactured by Leica or Arcturus, that permit laser capture microdissection of specific cells or cell groups from tissues were developed to solve this problem. To take full advantage of these instruments, however, one must be able to recognize cell populations of interest and, after they are harvested, to extract intact, unmodified RNA from them. Here we describe a novel, fast and simple method to fix and immunostain tissue sections that permits this to be done.
Xiang
Charlie C
CC
Laboratory of Genetics, National Institute of Mental Health, National Institute of Neurological Disorders and Stroke, NIH, 36 Convent Drive, Bethesda, MD 20892, USA.
Mezey
Eva
E
Chen
Mei
M
Key
Sharon
S
Ma
Li
L
Brownstein
Michael J
MJ
D000818:Animals; D001921:Brain / Q000166:cytology / Q000378:metabolism; D001923:Brain Chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry*; D005404:Fixatives / Q000737:chemistry*; D020869:Gene Expression Profiling; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D042282:Microdissection / Q000295:instrumentation; D041681:NIH 3T3 Cells; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000032:analysis / Q000302:isolation & purification*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013388:Succinimides / Q000737:chemistry*
D003432:Cross-Linking Reagents; D005404:Fixatives; D013388:Succinimides; D012313:RNA; C011240:dithiobis(succinimidylpropionate)