DNA cytometry and cytology by quantitative fluorescence image analysis in symptomatic bladder cancer patients

Abstract

A semi-automated quantitative fluorescence image analysis (QFIA) technique was developed with the Leitz TAS-Plus to detect bladder cancer using hyperploidy in urinary cells. Absolute nuclear fluorescence intensity (ANFI) (emission at 540 nm with excitation at 436 nm) of individual acridine-orange-stained cells was quantitated using (1) QFIA and (2) simple filter microspectrofluorophotometry (SFM). Both methods employed an internal phosphor particle standard which, when once calibrated against the DNA content of normal cells, obviates the necessity of routinely calibrating against normal cells in each sample. Results of SFM and QFIA were compared with routine Papanicolaou (Pap) cytopathology, using histopathology as the diagnostic standard in 272 samples from 67 symptomatic patients. The sensitivities for detecting low-grade transitional-cell carcinoma were 86% for SFM, 76% for QFIA, and 33% for Pap cytology. QFIA and SFM were significantly more sensitive at detecting bladder cancer than was Pap (0.01 greater than p greater than 0.001). Comparison of sensitivity obtained with bladder washings and urine samples showed that noninvasively obtained urines can be used. ANFI also detected recurrent and precancerous bladder lesions and kidney, ureter, and prostate lesions. This approach may prove generally useful in quantifying biochemical and immunological probes and should be broadly applicable as a research tool for studying the relationship of biochemical markers in the pathogenesis of disease and as a test for cancer control.

The Drake Health Registry Study: Findings from fifteen years of continuous bladder cancer screening

“…PAP is sensitive to high-grade malignancies but is generally insensitive to low-grade tumors [Nelson, 1985]. Various studies suggest an overall sensitivity of 66% for voided urine PAP cytology across the range of grades and stages for transitional cell tumors [Hemstreet et al, 1986], and 3) Quantitative fluorescence image analysis (QFIA) [West, 1965;Bass et al, 1987;Farrow, 1990;Hemstreet et al, 1990] is an evolving technology for quantitating biomarkers in exfoliated cells. This methodology detects early precancerous changes by measuring the amount of DNA or other biomarkers present in exfoliated uroepithelial cells.…”

Alterations in phenotypic biochemical markers in bladder epithelium during tumorigenesis.

“…Because quantitative rather than qualitative differences in gene expression and protein levels probably are responsible for most of the differences between malignant and normal phenotypes (13), quantitative fluorescence image analysis was used to quantify a panel of phenotypic markers in single cells from the biopsy specimens. Because of its visual morphologic component, quantitative fluorescence image analysis can link conventional morphologic assessment with quantitative biochemical markers at the single-cell level (14)(15)(16). The markers included quantitative fluorescence image analysis cytology (14), a combination of visual morphologic classification and identifying cells with DNA in excess of 5C (2C = amount of signal equivalent to the diploid chromosome complement), which is a marker for genetic instability; the p300 tumor-related antigen detected by the M344 monoclonal antibody (17); the differentiation-related proteins epidermal growth factor receptor (EGFR) (18,19) and G-actin (the globular monomeric precursor protein to actin filaments); and p185, the product of the HER-2/neu oncogene.…”

Occupational Hydrocarbon Exposure and Renal Histopathology

“…A useful extension of this study would be to utilize more sensitive testing to recognize acute nephrotoxic changes, potentially masked by repair mechanisms, that would help to define high-risk subsets or individual risk factors. This may be possible by utilizing quantitative fluorescence image analysis (Hemstreet et al, 1983;West et al, 1986;Bass et al, 1986). This technology could be important for the development of biological monitoring techniques to define high-risk groups analogous to the approach recently defined for bladder cancer Schulte et al, 1986).…”