Classification of fixed urological cells using Raman tweezers

Tim J. Harvey, Caryn Hughes, Andrew D. Ward, Elsa Correia Faria, Alex Henderson, Noel W. Clarke, Mick D. Brown, Richard D. Snook, Peter Gardner, Elsa Correia Faria - bill

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    Abstract

    In this paper we report on preliminary investigations into using Raman tweezers to classify urological cell lines. This builds on earlier work within the group, whereby Raman tweezer methodologies were developed, and the application of this technique to differentiate between live prostate cancer (CaP) and bladder cells lines (PC.-3 and MCiH-U1 respectively) was denion-In this present study we analysed chemically fixed cells using two different fixative methods; SurePalh™ (a commercial available liquid based cytology media) and 4% v/v formalin/PBS fixatives. The study has been expanded from our previous live cell study to include the androgen sensitive CaP cell line LNCaP, primary benign prostate hyperplasia (BPH) cells as well as primary urethral cells. Raman light from the cells was collected using a 514.5 nm Ar-ion laser excitation source in back-scattering configuration mode. Principal component-linear discriminate analysis (PC-LDA) models of resulting cell spectra were generated and these were validated using a blind comparison. Sensitivities and specificities of >72% and 90% respectively. for SurePalh fixed cells, and >9.1% and 98% respectively Tor 4% v/v formalin/PHS fixed cells was achieved. The higher prediction results for the formalin fixed cells can be attributed to a better signal-to-noise ratio for spectra obtained from these cells. Following on from this work, urological cell lines were exposed to urine for up to 12 hours to determine the effect of urine on the ability to classify these cells. Results indicate that urine has no detrimental effect on prediction results. © 2009 by WILEY-VCH Verlag GmbH & Co. KGaA.
    Original languageEnglish
    Pages (from-to)47-69
    Number of pages22
    JournalJournal of biophotonics
    Volume2
    Issue number1-2
    DOIs
    Publication statusPublished - Feb 2009

    Keywords

    • Cell fixation
    • Chemometrics
    • Cytology
    • Prostate cancer
    • Raman tweezers
    • Vibrational spectroscopy

    Research Beacons, Institutes and Platforms

    • Manchester Institute of Biotechnology

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