CARS hyperspectral imaging of cartilage aiming for state discrimination of cell

Manabu Shiozawa; Masataka Shirai; Junko Izumisawa; Maiko Tanabe; Koichi Watanabe

doi:10.1117/12.2208994

7 March 2016 CARS hyperspectral imaging of cartilage aiming for state discrimination of cell

Manabu Shiozawa, Masataka Shirai, Junko Izumisawa, Maiko Tanabe, Koichi Watanabe

Proceedings Volume 9704, Biomedical Vibrational Spectroscopy 2016: Advances in Research and Industry; 97040L (2016) https://doi.org/10.1117/12.2208994
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

Non-invasive cell analyses are increasingly important for medical field. A CARS microscope is one of the non-invasive imaging equipments and enables to obtain images indicating molecular distribution. Some studies on discrimination of cell state by using CARS images of lipid are reported. However, due to low signal intensity, it is still challenging to obtain images of the fingerprint region (800~1800 cm^-1), in which many spectrum peaks correspond to compositions of a cell. Here, to identify cell differentiation by using multiplex CARS, we investigated hyperspectral imaging of fingerprint region of living cells. To perform multiplex CARS, we used a prototype of a compact light source, which consists of a microchip laser, a single-mode fiber, and a photonic crystal fiber to generate supercontinuum light. Assuming application to regenerative medicine, we chose a cartilage cell, whose differentiation is difficult to be identified by change of the cell morphology. Because one of the major components of cartilage is collagen, we focused on distribution of proline, which accounts for approximately 20% of collagen in general. The spectrum quality was improved by optical adjustments about power branching ratio and divergence of broadband Stokes light. Hyperspectral images were successfully obtained by the improvement. Periphery of a cartilage cell was highlighted in CARS image of proline, and this result suggests correspondence with collagen generated as extracellular matrix. A possibility of cell analyses by using CARS hyperspectral imaging was indicated.

Citation Download Citation

Manabu Shiozawa, Masataka Shirai, Junko Izumisawa, Maiko Tanabe, and Koichi Watanabe "CARS hyperspectral imaging of cartilage aiming for state discrimination of cell", Proc. SPIE 9704, Biomedical Vibrational Spectroscopy 2016: Advances in Research and Industry, 97040L (7 March 2016); https://doi.org/10.1117/12.2208994

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KEYWORDS

Cartilage

Hyperspectral imaging

Collagen

Raman spectroscopy

Signal to noise ratio

Fiber lasers

Light

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