Flow Cytometry in Bladder Cancer Detection and Evaluation Using Acridine Orange Metachromatic Nucleic Acid Staining of Irrigation Cytology Specimens

Abstract

A new technique for simultaneous multiparameter deoxyribonucleic acid, ribonucleic acid and nuclear size measurements by flow cytometry was applied to the examination of bladder irrigation cytology specimens from 107 urologic patients. The cell samples from patients with bladder carcinoma could be distinguished from normal by 2 features: 1) an increase in the proportion of bladder epithelial cells with more than diploid deoxyribonucleic acid and 2) aneuploid cell peaks. These criteria identified 12 of 13 cases of invasive carcinoma, 24 of 28 cases of carcinoma in situ and 11 of 13 cases of papillary carcinoma. An increased proportion of cells with more than diploid deoxyribonucleic acid or aneuploidy was found in 9 of 14 patients with papilloma and 6 of 19 patients with a history of bladder tumors but no evident disease at present--these were believed owing to increased epithelial proliferative rates or nuclear chromatin abnormalities not visible by light microscopy. None of the 20 patients who had never had bladder tumors was abnormal. While the results in this small clinical trial have been most encouraging an additional descriptor of nuclear chromatin structure is believed necessary to discriminate benign, reactive proliferative epithelium from neoplasm when the latter is near diploid or shedding few cells. Studies to develop such a parameter presently are under way.

Flow Cytometric DNA/ Cytokeratin Analysis of Bladder Lavage: Methodical Aspects and Clinical Implications

“…A total rate of diagnosis of 81 % was achieved in a study by Collste et al [20]. This consists of 35% proven aneu ploidies (which is comparable to our own results), 5% with a CV of greater than 10% (an exclusion criterion in our investigation), 18% with a clear tetraploid peak as well as 23% with a proliferative rateof>l 5% S + G2 M phase.…”

“…Boileau et al [11] made the criticism that aneuploidy rates in previous studies by other authors [20] are created against nomenclature. They already see 'asymmetric peaks of Goi or simply just broadened CV' as proof of aneuploidy, thereby increasing sensitivity for well and moderately differentiated tumors without examining a control group of tumor-free patients by the same criteria.…”

“…Boileau et al [11] show that for proof of two extra chromosomes (corre sponding to 4% of DNA content) a CV of less than 2% is necessary, a rate of accuracy which is hardly obtainable by any flow cytometer in clinical specimens. The average CV value of measurements is usually not given or estimated to be between 6 and 7% [11,20]. Thus only a marked chromosome change is registered.…”

Dual-Parameter Immunoflow Cytometry in Diagnosis and Follow-Up of Patients with Bladder Cancer

“…Some investigators have addressed this problem. Collste et al [22]stained urothelial cells with acridine orange and reported that granulocytes could be distinguished from urothelial cells by lower DNA content, as well as by an increased nuclear size. Lymphocytes and monocytes, however, cannot be excluded by this technique.…”

Dual-Parameter Immunoflow Cytometry – A New Technique in the Noninvasive Diagnosis of Bladder Cancer

“…So far, some investigators have addressed this problem. Collste et al [13], staining urothelial cells with acridine orange, re ported that granulocytes could be distinguished from transitional cells by their low DNA, as well as by an increased nuclear size due to the presence of unfolded nuclei. Lymphocytes and monocytes, however, cannot be excluded by this technique.…”