A Prospective Comparison of DNA Quantitation by Image and Flow Cytometry

Abstract

Advances in computer and video technology suggest that image analysis may be practical method of measuring DNA that also allows visual confirmation of cell type. The purpose of this study was to prospectively compare DNA quantitation from 92 solid tumors in which DNA indices had been measured by image analysis of touch preparations (CAS 100) and flow cytometry of cell suspensions (FACScan). For 81 cases, there was excellent correlation between the two methods. For nine cases, however, an aneuploid population, usually near tetraploid, was identified by image but not by flow cytometry. Three cases had aneuploid peaks by flow cytometry that were not identified by image. Although these methods show good correlation, rare populations may be missed by CAS, presumably because of sampling errors in the touch preparation. Aneuploid populations may also be missed by flow cytometry, either because of cell loss during processing or because visual identification by image can increase sensitivity.

Development of a Flow Cytometric Method to Determine DNA Ploidy of Oesophageal Cancer Cells Obtained by Forceps Biopsy Samples During Oesophago-Gastro-Duodenoscopy

“…In concordance with other studies, the results in table 2 demonstrate that the sensitivity for detection of aneuploidy is higher with image analysis than with single labelling flow cytometry [13][14][15][16][17]. There are several explanations for these findings.…”

“…For determination of DNA content, image analysis is more established than single labelling flow cytometry and has a higher sensitivity for detecting aneuploid tumour cells [13][14][15][16][17]. For this method, only 250 tumour cell nuclei are needed and stromal cells can be excluded.…”

“…At the least, tumour cells are diluted by a large amount of stromal cells. For image analysis the cells must be separated manually by an experienced pathologist [13][14][15][16][17]. Therefore, this method is time consuming.…”

“…Therefore, this method is time consuming. Flow cytometry is faster, but the sensitivity for detection of small amounts of aneuploid tumour cells is lower [13][14][15][16][17]. The aim of our study was to establish a flow cytometric method in order to pre-operatively determine the DNA content of oesophageal cancer cells from biopsy samples obtained during oesophago-gastro-duodenoscopy.…”

Optimization of immunofluorescence methods by quantitative image analysis.

“…More recently, however, TV-based image analysis has been used to quantify the intensity of immunostaining and to eliminate subjective selection of thresholds. This technique has been used to quantify DNA content (Bauer et al, 1990), cell proliferation (Bacus et al, 1989) and protein expression (Baddoura et al, 1991;Bacus et al, 1988Bacus et al, ,1990.…”