Label-free observation of tissues by high-speed stimulated Raman spectral microscopy and independent component analysis

Ozeki, Yasuyuki; Otsuka, Yoichi; Sato, Shuya; Hashimoto, Hiroyuki; Umemura, Wataru; Sumimura, Kazuhiko; Nishizawa, Norihiko; Fukui, Kiichi; Itoh, Kazuyoshi

doi:https://doi.org/10.1117/12.2002772

Label-free observation of tissues by high-speed stimulated Raman spectral microscopy and independent component analysis

Ozeki, Yasuyuki; Otsuka, Yoichi; Sato, Shuya et al.

Proceedings of SPIE - The International Society for Optical Engineering, 2013, 8588, 858806

Number of Access:1,035（2025-08-14 04:33 Counts）

Identifier to cite or link to this item: https://hdl.handle.net/11094/79144

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Title	Label-free observation of tissues by high-speed stimulated Raman spectral microscopy and independent component analysis
Creator	NRID 1000060437374 1000060437374 CreatorName Ozeki, Yasuyuki
	NRID 1000070756460 1000070756460 researchmap permalink yoichiotsuka yoichiotsuka Scopus Author ID 55181504000 55181504000 ORCID 0000-0003-2304-6637 0000-0003-2304-6637 CreatorName Otsuka, Yoichi
	CreatorName Sato, Shuya
	CreatorName Hashimoto, Hiroyuki
	CreatorName Umemura, Wataru
	NRID 1000000452475 1000000452475 CreatorName Sumimura, Kazuhiko
	NRID 1000030273288 1000030273288 CreatorName Nishizawa, Norihiko
	NRID 1000000311770 1000000311770 researchmap permalink read0068600 read0068600 CreatorName Fukui, Kiichi
	NRID 1000080113520 1000080113520 CreatorName Itoh, Kazuyoshi
Description	SPIE BiOS, 2013, San Francisco, California, United States
Description	Yasuyuki Ozeki, Yoichi Otsuka, Shuya Sato, Hiroyuki Hashimoto, Wataru Umemura, Kazuhiko Sumimura, Norihiko Nishizawa, Kiichi Fukui, Kazuyoshi Itoh, "Label-free observation of tissues by high-speed stimulated Raman spectral microscopy and independent component analysis," Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 858806 (22 February 2013); https://doi.org/10.1117/12.2002772
Abstract	We have developed a video-rate stimulated Raman scattering (SRS) microscope with frame-by-frame wavenumber tunability. The system uses a 76-MHz picosecond Ti:sapphire laser and a subharmonically synchronized, 38-MHz Yb fiber laser. The Yb fiber laser pulses are spectrally sliced by a fast wavelength-tunable filter, which consists of a galvanometer scanner, a 4-f optical system and a reflective grating. The spectral resolution of the filter is ~ 3 cm-1. The wavenumber was scanned from 2800 to 3100 cm-1 with an arbitrary waveform synchronized to the frame trigger. For imaging, we introduced a 8-kHz resonant scanner and a galvanometer scanner. We were able to acquire SRS images of 500 x 480 pixels at a frame rate of 30.8 frames/s. Then these images were processed by principal component analysis followed by a modified algorithm of independent component analysis. This algorithm allows blind separation of constituents with overlapping Raman bands from SRS spectral images. The independent component (IC) spectra give spectroscopic information, and IC images can be used to produce pseudo-color images. We demonstrate various label-free imaging modalities such as 2D spectral imaging of the rat liver, two-color 3D imaging of a vessel in the rat liver, and spectral imaging of several sections of intestinal villi in the mouse. Various structures in the tissues such as lipid droplets, cytoplasm, fibrous texture, nucleus, and water-rich region were successfully visualized.
Publisher	SPIE
Source Title	Proceedings of SPIE - The International Society for Optical Engineering
Volume (Issue)	8588
Page	858806
Date of Issued	2013-02-22
Language	English
Handle URL	https://hdl.handle.net/11094/79144
PISSN	0277786X
NCID	AA10619755
Relation.isIdenticalTo	DOI (Publisher Version) https://doi.org/10.1117/12.2002772
Access Rights	open access
CopyRight	Copyright 2013 Society of Photo Optical Instrumentation Engineers (SPIE). One print or electronic copy may be made for personal use only. Systematic reproduction and distribution, duplication of any material in this publication for a fee or for commercial purposes, or modification of the contents of the publication are prohibited.
oaire:version	Version of Record
Category	会議発表論文 Conference Paper

Resource Type	conference paper
Local Type	Conference Paper
DCMI Type	Text
DCTERMS.bibliographicCitation	Proceedings of SPIE - The International Society for Optical Engineering. 2013, 8588, p. 858806
DC.title	Label-free observation of tissues by high-speed stimulated Raman spectral microscopy and independent component analysis
DC.creator	Ozeki, Yasuyuki
	Otsuka, Yoichi
	Sato, Shuya
	Hashimoto, Hiroyuki
	Umemura, Wataru
	Sumimura, Kazuhiko
	Nishizawa, Norihiko
	Fukui, Kiichi
	Itoh, Kazuyoshi
DC.publisher	SPIE
DC.language' scheme='DCTERMS.RFC1766	English
DC.type' scheme='DCTERMS.DCMIType	Text
DCTERMS.issued' scheme='DCTERMS.W3CDTF	2013-02-22
DC.identifier	https://doi.org/10.1117/12.2002772
DC.description	SPIE BiOS, 2013, San Francisco, California, United States
DC.description	Yasuyuki Ozeki, Yoichi Otsuka, Shuya Sato, Hiroyuki Hashimoto, Wataru Umemura, Kazuhiko Sumimura, Norihiko Nishizawa, Kiichi Fukui, Kazuyoshi Itoh, "Label-free observation of tissues by high-speed stimulated Raman spectral microscopy and independent component analysis," Proc. SPIE 8588, Multiphoton Microscopy in the Biomedical Sciences XIII, 858806 (22 February 2013); https://doi.org/10.1117/12.2002772
DCTERMS.abstract	We have developed a video-rate stimulated Raman scattering (SRS) microscope with frame-by-frame wavenumber tunability. The system uses a 76-MHz picosecond Ti:sapphire laser and a subharmonically synchronized, 38-MHz Yb fiber laser. The Yb fiber laser pulses are spectrally sliced by a fast wavelength-tunable filter, which consists of a galvanometer scanner, a 4-f optical system and a reflective grating. The spectral resolution of the filter is ~ 3 cm-1. The wavenumber was scanned from 2800 to 3100 cm-1 with an arbitrary waveform synchronized to the frame trigger. For imaging, we introduced a 8-kHz resonant scanner and a galvanometer scanner. We were able to acquire SRS images of 500 x 480 pixels at a frame rate of 30.8 frames/s. Then these images were processed by principal component analysis followed by a modified algorithm of independent component analysis. This algorithm allows blind separation of constituents with overlapping Raman bands from SRS spectral images. The independent component (IC) spectra give spectroscopic information, and IC images can be used to produce pseudo-color images. We demonstrate various label-free imaging modalities such as 2D spectral imaging of the rat liver, two-color 3D imaging of a vessel in the rat liver, and spectral imaging of several sections of intestinal villi in the mouse. Various structures in the tissues such as lipid droplets, cytoplasm, fibrous texture, nucleus, and water-rich region were successfully visualized.
DC.rights	Copyright 2013 Society of Photo Optical Instrumentation Engineers (SPIE). One print or electronic copy may be made for personal use only. Systematic reproduction and distribution, duplication of any material in this publication for a fee or for commercial purposes, or modification of the contents of the publication are prohibited.
DC.format	79144
citation_title	Label-free observation of tissues by high-speed stimulated Raman spectral microscopy and independent component analysis
citation_author	Ozeki, Yasuyuki
	Otsuka, Yoichi
	Sato, Shuya
	Hashimoto, Hiroyuki
	Umemura, Wataru
	Sumimura, Kazuhiko
	Nishizawa, Norihiko
	Fukui, Kiichi
	Itoh, Kazuyoshi
citation_publisher	SPIE
citation_language	English
citation_date	2013-02-22
citation_journal_title	Proceedings of SPIE - The International Society for Optical Engineering
citation_volume	8588
citation_firstpage	858806
citation_doi	https://doi.org/10.1117/12.2002772
citation_public_url	https://hdl.handle.net/11094/79144

Label-free observation of tissues by high-speed stimulated Raman spectral microscopy and independent component analysis

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