Preliminary micro-raman images of normal and malignant human skin cells

Michael A. Short; Harvey Lui; David I. McLean; Haishan Zeng; Michael X. Chen

doi:10.1117/12.647289

27 February 2006 Preliminary micro-Raman images of normal and malignant human skin cells

Michael A. Short, Harvey Lui, David I. McLean, Haishan Zeng, Michael X. Chen

Proceedings Volume 6093, Biomedical Vibrational Spectroscopy III: Advances in Research and Industry; 60930E (2006) https://doi.org/10.1117/12.647289
Event: SPIE BiOS, 2006, San Jose, California, United States

Abstract

Micro-Raman spectroscopy covering a frequency range from 200 to 4000 cm^-1 was used to image human skin melanocytes and keratinocytes with a spatial resolution of 0.5 μm. The cells were either cultivated on glass microscope slides or were located within thin sections of skin biopsies mounted on low fluorescence BaF₂. A commercially available system was used to obtain the spectra utilizing a x100 long working distance objective with a numerical aperture of 0.8, and a cooled CCD. Both 633 and 515 nm excitations were tried, although the latter proved to be more effcient at producing Raman emission mostly due to the 1/λ⁴ dependence in light scattering. Fluorescence emission from the cells was surprisingly low. The excitation power at the sample was kept below about 2 mW to avoid damaging the cells; this was the limiting factor on how quickly a Raman image could be obtained. Despite this diffculty we were able to obtain Raman images with rich information about the spectroscopic and structural features within the cytoplasm and cell nuclei. Differences were observed between the Raman images of normal and malignant cells. Spectra from purified DNA, RNA, lipids, proteins and melanin were obtained and these spectra were compared with the skin cell spectra with the aim of understanding how they are distributed over a cell and how the distribution changes between different cells.

Citation Download Citation

Michael A. Short, Harvey Lui, David I. McLean, Haishan Zeng, and Michael X. Chen "Preliminary micro-Raman images of normal and malignant human skin cells", Proc. SPIE 6093, Biomedical Vibrational Spectroscopy III: Advances in Research and Industry, 60930E (27 February 2006); https://doi.org/10.1117/12.647289

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