Investigation for the differentiation process of mouse ES cells by Raman spectroscopy

Yoshinori Yamaguchi; Maha A. El-Hagrasy; Eiichi Shimizu; Masato Saito; Eiichi Tamiya

doi:10.1117/12.907586

9 February 2012 Investigation for the differentiation process of mouse ES cells by Raman spectroscopy

Yoshinori Yamaguchi, Maha A. El-Hagrasy, Eiichi Shimizu, Masato Saito, Eiichi Tamiya

Proceedings Volume 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X; 82250J (2012) https://doi.org/10.1117/12.907586
Event: SPIE BiOS, 2012, San Francisco, California, United States

Abstract

The arrangement of differentiated pluripotent embryonic stem cells into three-dimensional aggregates, which are known as embryonic bodies, is a main step for progressing the embryonic stem cells differentiation. In this work, embryonic stem cells that were directly produced from the hanging drop step as a three-dimensional structure with no further twodimensional differentiation were diagnosed with Raman spectroscopy as a non-invasive and label-free technique. Raman spectroscopy was employed to discriminate between mouse embryonic bodies of different degrees of maturation. EBs were prepared applying the hanging drop method. The Raman scattering measurements were obtained in vitro with a Nanophoton RAMAN-11 micro-spectrometer (Japan: URL: www.nanophoton.jp equipped with an Olympus XLUM Plan FLN 20X/NA= 1.0 objective lens. Spectral data were smoothed, baseline corrected and normalized to the a welldefined intense 1003 cm-1 band (phenylalanine) which is insensitive to changes in conformation or environment. The differentiation process of embryonic stem cells is initiated by the removal of LIF from culture medium. 1, 7 and 17-dayold embryonic stem cells were collected and investigated by Raman spectroscopy. The main differences involve bands which decreased with maturation such as: 784 cm-1 (U, T, C ring br DNA/RNA, O-P-O str); 1177 cm-1 (cytosine, guanine) and 1578 cm-1 (G, A). It was found that with the progress of differentiation the protein content was amplified. The increase of protein to nucleic acid ratio was also previously observed with the progress of the differentiation process. Raman spectroscopy has the potential to distinguish between the Raman signatures of live embryonic stem cells with different degrees of maturation.

Citation Download Citation

Yoshinori Yamaguchi, Maha A. El-Hagrasy, Eiichi Shimizu, Masato Saito, and Eiichi Tamiya "Investigation for the differentiation process of mouse ES cells by Raman spectroscopy", Proc. SPIE 8225, Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues X, 82250J (9 February 2012); https://doi.org/10.1117/12.907586

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available

Members: $17.00

Non-members: $21.00 ADD TO CART

PROCEEDINGS
8 PAGES

DOWNLOAD PAPER SAVE TO MY LIBRARY

GET CITATION

RIGHTS & PERMISSIONS

Get copyright permission Get copyright permission on Copyright Marketplace

KEYWORDS

Raman spectroscopy

Proteins

Stem cells

Calcium

Laser induced fluorescence

Objectives

Imaging spectroscopy

Show All Keywords

Keywords/Phrases

Search In:

Publication Years