Importance of substrate and photo-induced effects in Raman spectroscopy of single functional erythrocytes

J Biomed Opt. 2003 Apr;8(2):173-8. doi: 10.1117/1.1559730.

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 (lambda=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.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Cell Culture Techniques / methods*
  • Cells, Immobilized
  • Erythrocytes / chemistry*
  • Erythrocytes / metabolism*
  • Feasibility Studies
  • Hemoglobins / chemistry*
  • Hemoglobins / metabolism*
  • Humans
  • Oxygen / chemistry
  • Oxygen / metabolism
  • Oxygen Consumption / physiology
  • Polylysine / chemistry
  • Spectrum Analysis, Raman / methods*

Substances

  • Hemoglobins
  • Polylysine
  • Oxygen