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Journal of Biomedical Optics

Importance of substrate- and photo-induced effects in Raman spectroscopy of single functional erythrocytes
Author(s): Kerstin K. Ramser; Erik J. Bjerneld; Camilla Fant; Mikael Kaell
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Paper Abstract

Hemoglobin (Hb) in single erythrocytes (red blood cells), adsorbed on polylysine-coated glass surfaces, was studied using resonance Raman spectroscopy and global Raman imaging. The erythrocytes were found to be sensitive to both surface adsorption and laser illumination. Substrate-dependent changes of the cell membrane shape were observed immediately after cell adsorption, while a photo-induced increase of fluorescence was observed for visible excitation (λ=514.5 nm). Concurrent changes in Raman spectra revealed a conversion of oxy-Hb (2+) to the inactive met-Hb state (3+). These effects severely complicate the interpretation of Raman images. However, at a low accumulated photon dose, the preparation method enabled the recording of Raman spectra during the oxygenation cycle of a single erythrocyte in buffer, which illustrates the feasibility of Raman investigations of functional cells in in-vivo environments.

Paper Details

Date Published: 1 April 2003
PDF: 6 pages
J. Biomed. Opt. 8(2) doi: 10.1117/1.1559730
Published in: Journal of Biomedical Optics Volume 8, Issue 2
Show Author Affiliations
Kerstin K. Ramser, Chalmers Univ. of Technology (Sweden)
Erik J. Bjerneld, Chalmers Univ. of Technology (Sweden)
Camilla Fant, Goeteborg Univ. (Sweden)
Mikael Kaell, Chalmers Univ. of Technology (Sweden)


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