Image analysis confirmation of DNA aneuploidy in flow cytometric DNA distributions having a wide coefficient of variation of the G0/G1 peak.
With nuclei extracted from paraffin-embedded tissues, resolution of normal diploid DNA and abnormal near-diploid/aneuploid populations by flow cytometry (FCM) is especially difficult. These samples, compared with fresh tissue, tend to show a broader DNA distribution, appearing as a wide (high) coefficient of variation (CV) of the G0/G1 peak. To address the question of whether there may be aneuploid populations hidden in wide CV diploid G0/G1 peaks, the authors measured DNA content in nuclei extracted from paraffin-embedded tumor tissue by morphometric image analysis (IA) in addition to FCM. Of 29 samples showing little evidence of DNA aneuploidy by FCM, in 20 of 20 with G0/G1 CVs greater than or equal to 5.50% there was an aneuploid population when analysis was performed by IA. Of the remaining nine samples with CVs less than or equal to 4.41%, all were diploid in the G0/G1 region by both FCM and IA. The presence of aneuploid populations in FCM distributions with wide CV G0/G1 peaks can be confirmed by IA.
McFadden
P W
PW
Department of Pathology, Medical College of Wisconsin, Milwaukee.
Clowry
L J
LJ
Daehnert
K
K
Hause
L L
LL
Koethe
S M
SM
D000782:Aneuploidy; D004273:DNA, Neoplasm / Q000032:analysis* / Q000235:genetics; D003720:Densitometry; D004171:Diploidy; D005434:Flow Cytometry / Q000379:methods; D006801:Humans; D007091:Image Processing, Computer-Assisted; D009369:Neoplasms / Q000032:analysis* / Q000235:genetics; D012044:Regression Analysis
D004273:DNA, Neoplasm