Kentaro Mochizuki, Yasuaki Kumamoto, Shunsuke Maeda, Masato Tanuma, Atsushi Kasai, Masashi Takemura, Yoshinori Harada, Hitoshi Hashimoto, Hideo Tanaka, Nicholas Isaac Smith, Katsumasa Fujita (2023) High-throughput line-illumination Raman microscopy with multislit detection., Biomedical optics express, Volume 14, Number 3, pp. 1015-1026
Published in 2023 Mar 1 (Electronic publication in Feb. 7, 2023, midnight )
(Abstract) Raman microscopy is an emerging tool for molecular imaging and analysis of living samples. Use of Raman microscopy in life sciences is, however, still limited because of its slow measurement speed for spectral imaging and analysis. We developed a multiline-illumination Raman microscope to achieve ultrafast Raman spectral imaging. A spectrophotometer equipped with a periodic array of confocal slits detects Raman spectra from a sample irradiated by multiple line illuminations. A comb-like Raman hyperspectral image is formed on a two-dimensional detector in the spectrophotometer, and a hyperspectral Raman image is acquired by scanning the sample with multiline illumination array. By irradiating a sample with 21 simultaneous illumination lines, we achieved high-throughput Raman hyperspectral imaging of mouse brain tissue, acquiring 1108800 spectra in 11.4 min. We also measured mouse kidney and liver tissue as well as conducted label-free live-cell molecular imaging. The ultrafast Raman hyperspectral imaging enabled by the presented technique will expand the possible applications of Raman microscopy in biological and medical fields.