Fourier Transform Infrared Spectroscopy as a Method for Monitoring the Molecular Dynamics of Lymphocyte Activation

Abstract

In this paper we report the application of Fourier transform infrared (FT-IR) microspectroscopy to monitor the molecular dynamics of lymphocyte activation. Infrared spectra of lymphocytes stimulated with the mitogen phytohaemagglutinin-L show spectral features 15 min after initial stimulation that are not apparent in resting lymphocytes. By analyzing the second-order derivatives of the raw spectra and applying principal components analysis (PCA), we conclude that the major spectral changes observed in the first hour result from an increase in overall RNA synthesis. Bands characteristic of RNA at 1244, 1080, 1050, 970, 1160, and 1120 cm−1 appear progressively more intense over time in the spectra of activated lymphocytes. The magnitude of these changes increases over time as the cell differentiates into a blast cell. The sensitivity of infrared spectroscopy to RNA moieties and the rapidity of the technique suggest a possible future role for FT-IR spectroscopy in histocompatibility testing.