Red blood cell Raman microscopy: modelling sub-cellular biochemistry

Abstract

We develop a quantitative Raman microscopy approach to study erythrocyte biochemistry at the sub-cellular level. To model Raman microscopy images, we review theory of Raman tensors and derive expressions for Raman responses suitable to compute Raman micro-images accounting effects of radial and vertical deformations of cellular envelopes. In application to membrane components, we extend the approach to a “counted per rotation” fast imaging protocol: once having Raman tensors for a molecule, precomputed expressions of molecular distributions can be used to construct Raman images of the modelled membrane envelope and its Raman spectra under any polarisation setting instantly. Using the theory, we review sub-cellular distributions of oxy-, deoxy- and methaemoglobins, as measured in experiment, considering their role in oxygen transport and oxidative stress mechanisms in the cytosol and when next to a membrane. We discuss possible applications of the approach in membrane specific studies, and its potential for combination with phase-sensitive and confocal fluorescence microscopy for advancing health care diagnostics.