Document Detail

Quantitative analysis of disseminated tumor cells in the bone marrow by automated fluorescence image analysis.
MedLine Citation:
PMID:  11135289     Owner:  NLM     Status:  MEDLINE    
Accurate quantification of disseminated tumor cells in hematological samples is of fundamental importance in clinical oncology. However, even highly standardized protocols allow only a rough estimation of the total analyzed cell number, as sample processing may have adverse effects on the number of cells available for analysis. The fluorescence-based microscopic scanning system (MetaCyte) detects, counts, captures, and relocates immunolabeled tumor cells in hematopoietic samples. We report on a cell-counting approach that has been implemented into the scanning system to precisely quantify the number of cells per slide. The cell-counting function, which was designed to determine the number of all nucleated (DAPI-stained) cells on the slide, allows an accurate counting of the tumor cells and the total number of cells analyzed in the given microscopic sample. The reliability of the cell-counting approach was demonstrated by the analysis of DAPI-stained images with 18-1,363 nucleated cells. A good correlation (r(2) = 0.965) between the manually and automatically gained results was observed. The counting accuracy could even be optimized after implementing a correction factor. To prove or disprove an interslide variation, routine bone marrow cytospin preparations from neuroblastoma patients were immunostained for GD2/FITC and counterstained with DAPI. Automatic cell counting of cytospin preparations from the same patients showed significant differences in the total cell number (up to 67% cell loss during preparation, with a maximum interslide difference of 4.7 x 10(5) mononuclear cells). We conclude that determination of the tumor cell content in hematopoietic samples is only reliable when it is performed together with accurate cell counting.
G Méhes; T Lörch; P F Ambros
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Publication Detail:
Type:  Comparative Study; Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Cytometry     Volume:  42     ISSN:  0196-4763     ISO Abbreviation:  Cytometry     Publication Date:  2000 Dec 
Date Detail:
Created Date:  2001-01-16     Completed Date:  2001-02-15     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  8102328     Medline TA:  Cytometry     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  357-62     Citation Subset:  IM    
CCRI, St. Anna Kinderspital, Vienna, Austria.
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MeSH Terms
Bone Marrow Cells / chemistry
Bone Marrow Neoplasms / pathology*,  secondary*
Cell Count*
Fluorescent Antibody Technique
Gangliosides / analysis,  immunology
Image Processing, Computer-Assisted
Microscopy, Fluorescence / methods*
Neuroblastoma / pathology
Reproducibility of Results
Reg. No./Substance:
0/Gangliosides; 65988-71-8/ganglioside, GD2

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