Laser-mediated microdissection for analysis of gene expression in synovial tissue.
In experimental rheumatology, transcriptomics is one of the most important methods for investigating the pathogenesis of diseases. The biological material of most studies on rheumatoid arthritis has been bulk rheumatoid synovial tissues, but they are not suitable because they consist of several kinds of cells or structures. Laser-mediated microdissection (LMM) is a useful tool for isolating particular cells from tissue specimen to assess the functions of each cell. The LMM system employs a combination of a microscope and a laser-beam generator to cut out target areas on cryosections. Tissue compartments or even a single viable cell can be isolated using a non-focused laser beam without direct contact to avoid contamination, and this process is called laser pressure catapulting. An ultraviolet-A laser enables target cells to be procured without any influence on the surrounding. This technique has already been used in several studies in rheumatology, and its validity has been confirmed. Combined with other new techniques such as real-time quantitative polymerase chain reaction or microarray analysis, LMM is becoming more important in the analysis of gene expression in rheumatology.
Hashimoto
Atsushi
A
Department of Rheumatology and Infectious Diseases, Kitasato University School of Medicine, 1-15-1 Kitasato, Sagamihara 228-8555, Japan. hashi@med.kitasato-u.ac.jp
Matsui
Toshimichi
T
Tanaka
Sumiaki
S
Ishikawa
Akira
A
Endo
Hirahito
H
Hirohata
Shunsei
S
Kondo
Hirobumi
H
Neumann
Elena
E
Tarner
Ingo Helmut
IH
Müller-Ladner
Ulf
U
D001172:Arthritis, Rheumatoid; D016616:Cryoultramicrotomy / Q000379:methods; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D042282:Microdissection / Q000295:instrumentation / Q000379:methods*; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D012333:RNA, Messenger / Q000032:analysis; D013583:Synovial Membrane / Q000378:metabolism*
D012333:RNA, Messenger