PMID- 8900478
OWN - NLM
STAT- MEDLINE
DCOM- 19961223
LR  - 20061115
IS  - 0196-4763 (Print)
IS  - 0196-4763 (Linking)
VI  - 23
IP  - 4
DP  - 1996 Apr 1
TI  - Genotype/phenotype analyses of low frequency tumor cells using computerize image 
      microscopy.
PG  - 344-9
AB  - Techniques to identify low frequency (i.e., 10(-4)-10(-5)) tumor cells in bone 
      marrow and peripheral blood of cancer patients provide opportunities for early 
      detection of disseminated disease and characterization of the properties of cells 
      released from the tumor. Low frequency epithelial cells in marrow and blood can 
      be detected using immunophenotypic markers directed against intracellular and/or 
      cell surface antigens. However, nonspecific phenotypic labeling may compromise 
      the ability to discriminate tumor from nontumor cells. We describe optimization 
      of slide-based approaches that facilitate identification and subsequent molecular 
      cytogenetic characterization of rare tumor cell populations in hematopoietic 
      tissues. Colon tumor cells seeded in a hematopoietic background provided a model 
      system to optimize methodologies that are applicable to detection and 
      quantification of micrometastases in clinical specimens. Mixtures of 
      cytogenetically aberrant epithelial cells and hematopoietic cells on slides were 
      labeled with an anti-cytokeratin 20 (anti-CK20) antibody that recognizes > 90% of 
      colon adenocarcinomas. Computerized image analysis was used to record the 
      location of the immunofluorescent cells on microscopic slides. Cells on slides 
      were then hybridized using fluorescence in situ hybridization (FISH) with 
      repeat-sequence DNA probes to detect aneusomies. Previously discriminated 
      epithelial cells were relocated for molecular cytogenetic characterization. Tumor 
      cells present at frequencies approximating 5 x 10(-5) were discriminated on the 
      basis of immunofluorescence and cell size. A low frequency (4 x 10(-4)) 
      population of normal hematopoietic cells also labeled with anti-CK20 (data not 
      shown). This strategy of sequential immunophenotyping and molecular cytogenetic 
      analyses may be useful to discriminate tumor from nontumor cells in cancer 
      patients with micrometastatic disease.
FAU - Litle, V R
AU  - Litle VR
AD  - Department of Surgery, University of California, San Francisco, USA.
FAU - Lockett, S J
AU  - Lockett SJ
FAU - Pallavicini, M G
AU  - Pallavicini MG
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Cytometry
JT  - Cytometry
JID - 8102328
RN  - 0 (Intermediate Filament Proteins)
RN  - 0 (KRT20 protein, human)
RN  - 0 (Keratin-20)
SB  - IM
MH  - Genotype
MH  - Humans
MH  - Image Processing, Computer-Assisted
MH  - Intermediate Filament Proteins/*immunology
MH  - Keratin-20
MH  - Microscopy, Fluorescence
MH  - Phenotype
MH  - Sensitivity and Specificity
MH  - Tumor Cells, Cultured
EDAT- 1996/04/01 00:00
MHDA- 2000/06/20 09:00
CRDT- 1996/04/01 00:00
PHST- 1996/04/01 00:00 [pubmed]
PHST- 2000/06/20 09:00 [medline]
PHST- 1996/04/01 00:00 [entrez]
AID - 10.1002/(SICI)1097-0320(19960401)23:4<344::AID-CYTO11>3.0.CO;2-T [pii]
AID - 10.1002/(SICI)1097-0320(19960401)23:4<344::AID-CYTO11>3.0.CO;2-T [doi]
PST - ppublish
SO  - Cytometry. 1996 Apr 1;23(4):344-9. doi: 
      10.1002/(SICI)1097-0320(19960401)23:4<344::AID-CYTO11>3.0.CO;2-T.