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Title: Cell-Type-Specific Genome-wide Expression Profiling after Laser Capture Microdissection of Living Tissue

The purpose of this technical feasibility study was to develop and evaluate robust microgenomic tools for investigations of genome-wide expression of very small numbers of cells isolated from whole tissue sections. Tissues contain large numbers of cell-types that play varied roles in organ function and responses to endogenous and exogenous toxicants whether bacterial, viral, chemical or radiation. Expression studies of whole tissue biopsy are severely limited because heterogeneous cell-types result in an averaging of molecular signals masking subtle but important changes in gene expression in any one cell type(s) or group of cells. Accurate gene expression analysis requires the study of specific cell types in their tissue environment but without contamination from surrounding cells. Laser capture microdissection (LCM) is a new technology to isolate morphologically distinct cells from tissue sections. Alternative methods are available for isolating single cells but not yet for their reliable genome-wide expression analyses. The tasks of this feasibility project were to: (1) Develop efficient protocols for laser capture microdissection of cells from tissues identified by antibody label, or morphological stain. (2) Develop reproducible gene-transcript analyses techniques for single cell-types and determine the numbers of cells needed for reliable genome-wide analyses. (3) Validate the technology for epithelialmore » and endothelial cells isolated from the gastrointestinal tract of mice.« less
Authors:
;
Publication Date:
OSTI Identifier:
15014606
Report Number(s):
UCRL-TR-209634
TRN: US200807%%738
DOE Contract Number:
W-7405-ENG-48
Resource Type:
Technical Report
Resource Relation:
Other Information: PBD: 9 Feb 2005
Research Org:
Lawrence Livermore National Lab., Livermore, CA (US)
Sponsoring Org:
US Department of Energy (US)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; BIOPSY; CONTAMINATION; GASTROINTESTINAL TRACT; GENES; LASERS; MICE; ORGANS