Publication: Advances in isolation and
characterization of homogeneous cell
populations using laser microdissection
Authors
Mizuaral, S. ; Takahashi, K. ; Kobayashi, T. ; Kotani, H.
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Publisher
Murcia : F. Hernández
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DOI
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info:eu-repo/semantics/article
Description
Abstract
The isolation and characterization of
homogeneous cell populations are of great importance
for the analysis of gene expression, because normal
tissues contain various types of cells, and the differences
in the populations of isolated cells exert significant
effects on gene expression analysis. Researchers have
attempted to develop methods for the isolation of
homogeneous cell populations, such as flow cytometry
and mechanical dissection. However, the recent
emergence of laser-assisted microdissection has
revolutionized the isolation of single-cell populations
from solid tissues. With the help of a cutting laser, laser
microdissection can isolate tissues (cells) of interest
without contamination from surrounding tissues with the
microscopic visualization field. By combining laser
microdissection and subsequent microarray technology,
several studies have resulted in the identification of
disease-related genes. In this review, we summarize the
principle of laser microdissection and provide several
successful examples of target-gene identification using
the conventional method combining laser
microdissection and microarray. Next, we discuss the
practical drawbacks of the combinational method, such
as the need for a large number of cells and the
disturbance of the relative abundance of transcripts
during RNA amplification. We introduce our
modifications to combined laser microdissection and
microarray for detection of disease-related genes; the
technique is simple, yet practical and accurate. Finally,
versatile applications of laser microdissection, not only
to transcript expression analysis, but also to other
genomics and proteomics analyses are, also presented.
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