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Proceedings Paper

Feasibility of Raman spectroscopy in vitro after 5-ALA-based fluorescence diagnosis in the bladder
Author(s): M. C. M. Grimbergen; C. F. P. van Swol; R. J. A. van Moorselaar; A, Mahadevan-Jansen; N. Stone
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Paper Abstract

Photodynamic diagnosis (PDD) has become popular in bladder cancer detection. Several studies have however shown an increased false positive biopsies rate under PDD guidance compared to conventional cystoscopy. Raman spectroscopy is an optical technique that utilizes molecular specific, inelastic scattering of light photons to interrogate biological tissues, which can successfully differentiate epithelial neoplasia from normal tissue and inflammations in vitro. This investigation was performed to show the feasibility of NIR Raman spectroscopy in vitro on biopsies obtained under guidance of 5-ALA induced PPIX fluorescence imaging. Raman spectra of a PPIX solution was measured to obtain a characteristic signature for the photosensitzer without contributions from tissue constituents. Biopsies were obtained from patients with known bladder cancer instilled with 50ml, 5mg 5-ALA two hours prior to trans-urethral resection of tumor (TURT). Additional biopsies were obtained at a fluorescent and non-fluorescent area, snap-frozen in liquid nitrogen and stored at -80 °C. Each biopsy was thawed before measurements (10sec integration time) with a confocal Raman system (Renishaw Gloucestershire, UK). The 830 nm excitation (300mW) source is focused on the tissue by a 20X ultra-long-working-distance objective. Differences in fluorescence background between the two groups were removed by means of a special developed fluorescence subtraction algorithm. Raman spectra from ALA biopsies showed different fluorescence background which can be effectively removed by a fluorescence subtraction algorithm. This investigation shows that the interaction of the ALA induced PPIX with Raman spectroscopy in bladder samples. Combination of these techniques in-vivo may lead to a viable method of optical biopsies in bladder cancer detection.

Paper Details

Date Published: 22 February 2006
PDF: 6 pages
Proc. SPIE 6078, Photonic Therapeutics and Diagnostics II, 60781D (22 February 2006); doi: 10.1117/12.655616
Show Author Affiliations
M. C. M. Grimbergen, Univ. Medical Ctr. Utrecht (Netherlands)
C. F. P. van Swol, Univ. Medical Ctr. Utrecht (Netherlands)
St. Antonius Hospital Nieuwegein (Netherlands)
R. J. A. van Moorselaar, Univ. Medical Ctr. Utrecht (Netherlands)
A, Mahadevan-Jansen, Vanderbilt Univ. (United States)
N. Stone, Gloucestershire Hospitals NHS Foundation Trust (United Kingdom)

Published in SPIE Proceedings Vol. 6078:
Photonic Therapeutics and Diagnostics II
Kenton W. Gregory; Nikiforos Kollias; Reza S. Malek; Michael D. Lucroy; Henry Hirschberg; Brian Jet-Fei Wong; Eugene A. Trowers; Werner T.W. de Riese; Justus F. R. Ilgner; Steen J. Madsen; Lloyd P. Tate; Haishan Zeng; Guillermo J. Tearney; Bernard Choi, Editor(s)

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