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Plant Cell Advance Online Publication Published on February 21, 2003; 10.1105/tpc.008102
Received September 30, 2002 Laser-Capture Microdissection, a Tool for the Global Analysis of Gene Expression in Specific Plant Cell Types: Identification of Genes Expressed Differentially in Epidermal Cells or Vascular Tissues of Maize
1
Department of Agronomy, Iowa State University, Ames, Iowa 50011; Graduate School
of Agricultural and Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo
113-8657, Japan
* To whom correspondence should be addressed. E-mail: schnable{at}iastate.edu.
Laser-capture microdissection (LCM) allows for the one-step procurement of large
homogeneous populations of cells from tissue sections. In mammals, LCM has been used
to conduct cDNA microarray and proteomics studies on specific cell types. However,
LCM has not been applied to plant cells, most likely because plant cell walls make
it difficult to separate target cells from surrounding cells and because ice crystals
can form in the air spaces between cells when preparing frozen sections. By fixing
tissues, using a cryoprotectant before freezing, and using an adhesive-coated slide
system, it was possible to capture large numbers (>10,000) of epidermal cells and
vascular tissues (vascular bundles and bundle sheath cells) from ethanol:acetic acid-fixed
coleoptiles of maize. RNA extracted from these cells was amplified with T7 RNA polymerase
and used to hybridize a microarray containing
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