Integral spectrophotometric analysis of 5-aminolaevulinic acid-induced fluorescence cytology of the urinary bladder

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Abstract

OBJECTIVE

To evaluate whether tumour cells can be detected in bladder lavage fluid samples by an objective spectrofluorometric method, as 5-aminolaevulinic acid (ALA)-induced fluorescence endoscopy (AFE) and cytology are promising valuable tools for detecting transitional cell carcinoma of the urinary bladder (TCCB).

MATERIALS AND METHODS

After instilling ALA into the urinary bladder, lavage samples were collected and their sediments analysed spectroscopically under blue excitation at ≈ 400 nm wavelength. During AFE, biopsies were taken. From 62 cases, 24 patients had a histologically confirmed TCCB (group A), 28 had a history of TCCB but no evidence of disease (group B) and 10 were negative for TCCB (group C).

RESULTS

Lavage sediments of all patients fluoresced in the green spectral range, typical of cellular autofluorescence. Sediments of all patients of group A caused red fluorescence peaking at 635 nm, indicating protoporphyrin IX (PPIX). The PPIX signals derived from bleaching spectra were significantly different between benign and malignant findings (P = 0.001). There was another red fluorescence peak at ≈ 620 nm; in 19 cases its intensity exceeded the intensity of the PPIX signal.

CONCLUSIONS

Spectrofluorometric analysis of lavage sample sediments can be used to detect tumour-associated red fluorescence of PPIX in TCCB. Immediate significant and objective measurements are possible, which could be further automated for the rapid diagnosis of TCCB.

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