Published online 16 December 2004
Nucleic Acids Research, Vol. 32 No. 22 © Oxford University Press 2004; all rights reserved
Using DSP, a reversible cross-linker, to fix tissue sections for immunostaining, microdissection and expression profiling
Laboratory of Genetics, National Institute of Mental Health and 1 Basic Neuroscience Program, National Institute of Neurological Disorders and Stroke, NIH, 36 Convent Drive, Bethesda, MD 20892, USA
* To whom correspondence should be addressed at Laboratory of Genetics, NIMH/NHGRI, NIH, Building 36, Room 3D06, 36 Convent Drive, Bethesda, MD 20892-4094, USA. Tel: +301 496 5351; Fax: +301 435 5465; Email: brownstm{at}mail.nih.gov
Received June 28, 2004; Revised October 7, 2004; Accepted November 29, 2004
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.
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