Optimisation of Wavelength Modulated Raman Spectroscopy: Towards High Throughput Cell Screening

In the field of biomedicine, Raman spectroscopy is a powerful technique to discriminate between normal and cancerous cells. However the strong background signal from the sample and the instrumentation affects the efficiency of this discrimination technique. Wavelength Modulated Raman spectroscopy (WMRS) may suppress the background from the Raman spectra. In this study we demonstrate a systematic approach for optimizing the various parameters of WMRS to achieve a reduction in the acquisition time for potential applications such as higher throughput cell screening. The Signal to Noise Ratio (SNR) of the Raman bands depends on the modulation amplitude, time constant and total acquisition time. It was observed that the sampling rate does not influence the signal to noise ratio of the Raman bands if three or more wavelengths are sampled. With these optimised WMRS parameters, we increased the throughput in the binary classification of normal human urothelial cells and bladder cancer cells by reducing the total acquisition time to 6 s which is significantly lower in comparison to previous acquisition times required for the discrimination between similar cell types.

[1]  Thomas R Huser,et al.  Raman spectroscopy and microscopy of individual cells and cellular components , 2008 .

[2]  M. Houlne,et al.  Spatially resolved analysis of small particles by confocal Raman microscopy: depth profiling and optical trapping. , 2004, Analytical chemistry.

[3]  Kishan Dholakia,et al.  Optimal algorithm for fluorescence suppression of modulated Raman spectroscopy. , 2010, Optics express.

[4]  T. B. Bakker Schut,et al.  Discriminating basal cell carcinoma from its surrounding tissue by Raman spectroscopy. , 2002, The Journal of investigative dermatology.

[5]  J. Masters,et al.  Human bladder cancer in vitro drug sensitivities: range and stability in long-term culture. , 1986, British Journal of Cancer.

[6]  Jürgen Popp,et al.  Tumour cell identification by means of Raman spectroscopy in combination with optical traps and microfluidic environments. , 2011, Lab on a chip.

[7]  Kishan Dholakia,et al.  Fluorescence suppression using wavelength modulated Raman spectroscopy in fiber-probe-based tissue analysis. , 2012, Journal of biomedical optics.

[8]  P R Carey,et al.  Raman Spectroscopy, the Sleeping Giant in Structural Biology, Awakes* , 1999, The Journal of Biological Chemistry.

[9]  G Rusciano,et al.  Enhancing Raman tweezers by phase-sensitive detection. , 2007, Analytical chemistry.

[10]  James J. Valentini,et al.  Chapter 1 – Coherent Anti-Stokes Raman Spectroscopy* , 1985 .

[11]  Alex Henderson,et al.  Spectral discrimination of live prostate and bladder cancer cell lines using Raman optical tweezers. , 2008, Journal of biomedical optics.

[12]  K. H. Levin,et al.  Wavelength‐modulation Raman spectroscopy , 1978 .

[13]  David J Brady,et al.  Multi-excitation Raman spectroscopy technique for fluorescence rejection. , 2008, Optics express.

[14]  R. Hochmuth Solid and liquid behavior of red cell membrane. , 1982, Annual review of biophysics and bioengineering.

[15]  M Mazilu,et al.  Dual beam fibre trap for Raman micro-spectroscopy of single cells. , 2006, Optics express.

[16]  G. Puppels,et al.  Towards oncological application of Raman spectroscopy , 2009, Journal of biophotonics.

[17]  F. L. Galeener FM spectroscopy: Raman scattering and luminescence , 1977 .

[18]  C. Reznikoff,et al.  Characterization of human uroepithelial cells immortalized in vitro by simian virus 40. , 1987, Cancer research.

[19]  Kishan Dholakia,et al.  Fluorescence suppression within Raman spectroscopy using annular beam excitation , 2007 .

[20]  Kishan Dholakia,et al.  Online fluorescence suppression in modulated Raman spectroscopy. , 2010, Analytical chemistry.

[21]  A. Sasso,et al.  Phase-sensitive detection in Raman tweezers , 2006 .

[22]  Pavel Matousek,et al.  Efficient Rejection of Fluorescence from Raman Spectra Using Picosecond Kerr Gating , 1999 .

[23]  D. Williams,et al.  CW Laser Wavelength Modulation in Raman and Site Selection Fluorescence Spectroscopy , 1976 .