PMID- 10874675
OWN - NLM
STAT- MEDLINE
DCOM- 20000926
LR  - 20061115
IS  - 0893-3952 (Print)
IS  - 0893-3952 (Linking)
VI  - 13
IP  - 6
DP  - 2000 Jun
TI  - Laser capture microdissection-guided fluorescence in situ hybridization and flow 
      cytometric cell cycle analysis of purified nuclei from paraffin sections.
PG  - 705-11
AB  - Laser capture microdissection (LCM) has recently been identified as a quick, 
      simple, and effective method by which microdissection of complex tissue specimens 
      for molecular analysis can be routinely performed. Assessment of gene copy number 
      by fluorescence in situ hybridization (FISH) is useful for the analysis of 
      molecular genetic alterations in cancer. Unfortunately, the application of FISH 
      to paraffin sections of tumor specimens is fraught with technical difficulty and 
      potential artifacts. Our results demonstrate that LCM-microdissected nuclei are 
      suitable for FISH gene copy analysis. Amplification of genes in cancer specimens 
      can be detected as easily in LCM-prepared nuclei as in fresh nuclei from cancer 
      tissue specimens. Furthermore, contamination of tumor specimens by normal cells 
      can make interpretation of flow cytometric cell cycle analysis difficult. Our 
      results show that LCM-microdissected nuclei can also be used for flow cytometric 
      cell cycle and ploidy analysis. LCM/FISH offers the advantages of multicolor FISH 
      in a morphologically defined cell population, without the technical problems of 
      FISH performed on paraffin sections. This technique should further simplify the 
      methodology required to perform copy number analysis of tumor suppressor or 
      protooncogenes in archived cancer specimens. The use of LCM specimens will also 
      improve the specificity and simplify the interpretation of flow cytometric cell 
      cycle and ploidy analysis of breast cancer specimens.
FAU - DiFrancesco, L M
AU  - DiFrancesco LM
AD  - Department of Pathology, University of Calgary, Alberta, Canada.
FAU - Murthy, S K
AU  - Murthy SK
FAU - Luider, J
AU  - Luider J
FAU - Demetrick, D J
AU  - Demetrick DJ
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Mod Pathol
JT  - Modern pathology : an official journal of the United States and Canadian Academy 
      of Pathology, Inc
JID - 8806605
SB  - IM
MH  - Breast Neoplasms/genetics/pathology
MH  - Cell Cycle
MH  - Cell Nucleus/*ultrastructure
MH  - *Flow Cytometry/methods
MH  - *Gene Dosage
MH  - Humans
MH  - *In Situ Hybridization, Fluorescence/methods
MH  - Paraffin Embedding
MH  - Ploidies
EDAT- 2000/06/30 11:00
MHDA- 2000/09/30 11:01
CRDT- 2000/06/30 11:00
PHST- 2000/06/30 11:00 [pubmed]
PHST- 2000/09/30 11:01 [medline]
PHST- 2000/06/30 11:00 [entrez]
AID - 10.1038/modpathol.3880120 [doi]
PST - ppublish
SO  - Mod Pathol. 2000 Jun;13(6):705-11. doi: 10.1038/modpathol.3880120.