Laser capture microdissection for analysis of single cells.
Laser capture microdissection (LCM) can be used to obtain single cells or a homogeneous population of cells for molecular analysis. This approach becomes even more powerful when it is combined with immunocytochemical staining using specific antibodies to label the cells of interest before LCM (referred to as immuno-LCM). These techniques have been applied in our laboratory to the analysis of pituitary cells from dissociated tissues and from cultured populations of heterogeneous pituitary, thyroid, and carcinoid tumor cells, as well as for the analysis of single cells in various sarcomas. When combined with reverse transcriptase polymerase chain reaction (RT-PCR) and Southern blot analysis, the sensitivity of this method is increased, allowing the reproducible analysis of gene expression from 1 to 10 cells. These methods show the utility of immuno-LCM as well as LCM combined with RT-PCR for cellular and molecular studies of gene expression.
Nakamura
Nobuki
N
Department of Laboratory Medicine and Pathology, Mayo Clinic Medical College, Rochester, MN, USA.
Ruebel
Katharina
K
Jin
Long
L
Qian
Xiang
X
Zhang
Heyu
H
Lloyd
Ricardo V
RV
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D007834:Lasers; D042282:Microdissection / Q000379:methods*; D010902:Pituitary Gland / Q000737:chemistry / Q000166:cytology; D012313:RNA / Q000032:analysis / Q000302:isolation & purification; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013961:Thyroid Gland / Q000737:chemistry / Q000166:cytology; D014407:Tumor Cells, Cultured
D012313:RNA