Immuno-laser capture microdissection of rat brain neurons for real time quantitative PCR.
Laser capture microdissection (LCM) is a technical approach for obtaining microscopic samples as small as individual cells from tissues for molecular analysis. While the principles and details of the operation of LCM instruments, the technical requirements for obtaining identified cells for LCM "picking", all share the common feature of using a laser in combination with a microscope to microdissect and remove cells from tissue slices (or cultured cells) mounted on a glass slide. The use of LCM is becoming widespread in pathology laboratories and is increasingly being used for gene expression studies in cell biology. The approach is particularly powerful when used in conjunction with immunostaining techniques to obtain enriched RNA samples from cells that have been collected by picking and gathering phenotypically similar cells from anatomically complex organs such as the brain. In the present chapter, we describe an approach for combining immunocytochemistry with LCM to obtain RNA for real time quantitative PCR.
Baskin
Denis G
DG
Division of Endocrinology/Metabolism, VA Puget Sound Health Care System, Seattle, WA, USA.
Bastian
L Scot
LS
D000818:Animals; D001921:Brain / Q000166:cytology*; D007150:Immunohistochemistry / Q000379:methods*; D007834:Lasers; D042282:Microdissection / Q000295:instrumentation / Q000379:methods*; D009474:Neurons / Q000737:chemistry* / Q000166:cytology; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000302:isolation & purification*; D051381:Rats; D016707:Tissue Fixation
D012333:RNA, Messenger