Deep-UV resonance Raman imaging of a cell (Conference Presentation)

Yasuaki Kumamoto

doi:10.1117/12.2236187

11 November 2016 Deep-UV resonance Raman imaging of a cell (Conference Presentation)

Yasuaki Kumamoto

Proceedings Volume 9926, UV and Higher Energy Photonics: From Materials to Applications; 992607 (2016) https://doi.org/10.1117/12.2236187
Event: SPIE Nanoscience + Engineering, 2016, San Diego, California, United States

Abstract

Raman microscopy enables a sensitive, label-free molecular imaging of cells. Employing deep-UV (DUV) light for Raman excitation allows selective measurement of nucleotide bases and aromatic amino acids in a cell, without spectral overlapping of components with a large quantity (i.e. lipid, peptide), because their Raman scattering are specifically enhanced due to the resonance effect. To implement DUV resonance Raman imaging of cells, I previously established a home-built Raman microscope equipped with a DUV laser (λ = 257.2 nm). Raman image representing the distribution of cellular nucleic acid can be reconstructed with the intensity of a Raman band selectively assigned to adenine and guanine. Unfortunately, DUV resonance Raman imaging of cells is severely hindered by molecular photodegradation that occurs after a molecule absorbs DUV light during Raman measurement, precluding a high signal-to-noise ratio and repetitive measurement. To address this issue, I developed a technique for molecular protection under DUV exposure; the trivalent ions of lanthanide group including terbium, europium, and thulium could significantly suppress the molecular photodegradation by relaxing the DUV-excited molecules. The buffer solution containing any of these lanthanide ions with the concentration of 100 µM or higher could provide less destruction of the cellular structures, including nucleotide bases, than the one without the ions, under DUV exposure. Utilizing such protective effects of the lanthanide ions, I successfully achieved a twice higher signal-to-noise ratio and repetitive DUV Raman imaging of cells.

Conference Presentation

Citation Download Citation

Yasuaki Kumamoto "Deep-UV resonance Raman imaging of a cell (Conference Presentation)", Proc. SPIE 9926, UV and Higher Energy Photonics: From Materials to Applications, 992607 (11 November 2016); https://doi.org/10.1117/12.2236187

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KEYWORDS

Deep ultraviolet

Raman spectroscopy

Ions

Lanthanides

Molecules

Signal to noise ratio

Microscopy

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