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Spectroscopic study of human lung epithelial cells (A549) in culture: Living cells versus dead cells

Spectroscopic study of human lung epithelial cells (A549) in culture: Living cells versus dead cells The noninvasive analysis of living cells grown on 3‐dimensional scaffold materials is a key point in tissue engineering. In this work we show the capability of Raman spectroscopy for use as a noninvasive method to distinguish cells at different stages of the cell cycle and living cells from dead cells. The spectral differences between cells in different stages of the cell cycle are characterized mainly by variations in DNA vibrations at 782, 788, and 1095 cm−1. The Raman spectrum of dead human lung derived (A549 line) cells indicates the breakdown of both phosphodiester bonds and DNA bases. The most sensitive peak for identifying dead cells is the 788 cm−1 peak corresponding to DNA O&bond;P&bond;O backbone stretching. The magnitude of this peak is reduced by 80% in the spectrum of dead cells. Changes in protein peaks suggest significant conformational changes; for example, the magnitude of the 1231 cm−1 peak assigned to random coils is reduced by 63% for dead cells. The sharp peak of phenylalanine at 1005 cm−1 drops to half, indicating a decrease of stable proteins associated with cell death. The differences in the 1190–1385 cm−1 spectral region also suggest a decrease in the amount of nucleic acids and proteins. Using curve fitting, we quantify these spectral differences that can be used as markers of cell death. © 2003 Wiley Periodicals, Inc. Biopolymers (Biospectroscopy), 2003 http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Biopolymers Wiley

Spectroscopic study of human lung epithelial cells (A549) in culture: Living cells versus dead cells

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References (26)

Publisher
Wiley
Copyright
Copyright © 2003 Wiley Periodicals, Inc., A Wiley Company
ISSN
0006-3525
eISSN
1097-0282
DOI
10.1002/bip.10378
pmid
12833477
Publisher site
See Article on Publisher Site

Abstract

The noninvasive analysis of living cells grown on 3‐dimensional scaffold materials is a key point in tissue engineering. In this work we show the capability of Raman spectroscopy for use as a noninvasive method to distinguish cells at different stages of the cell cycle and living cells from dead cells. The spectral differences between cells in different stages of the cell cycle are characterized mainly by variations in DNA vibrations at 782, 788, and 1095 cm−1. The Raman spectrum of dead human lung derived (A549 line) cells indicates the breakdown of both phosphodiester bonds and DNA bases. The most sensitive peak for identifying dead cells is the 788 cm−1 peak corresponding to DNA O&bond;P&bond;O backbone stretching. The magnitude of this peak is reduced by 80% in the spectrum of dead cells. Changes in protein peaks suggest significant conformational changes; for example, the magnitude of the 1231 cm−1 peak assigned to random coils is reduced by 63% for dead cells. The sharp peak of phenylalanine at 1005 cm−1 drops to half, indicating a decrease of stable proteins associated with cell death. The differences in the 1190–1385 cm−1 spectral region also suggest a decrease in the amount of nucleic acids and proteins. Using curve fitting, we quantify these spectral differences that can be used as markers of cell death. © 2003 Wiley Periodicals, Inc. Biopolymers (Biospectroscopy), 2003

Journal

BiopolymersWiley

Published: Jan 1, 2003

Keywords: Raman spectroscopy; cell death; cell cycle; tissue engineering

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