Imaging with Extrinsic Raman Labels

Abstract

In two separate examples we demonstrate the use of extrinsic Raman scattering probes for imaging of biological samples. First, the distribution of cholesterol in a rat eye lens is determined with the use of the Raman scattered light from filipin, a molecule which binds specifically to cholesterol. The protein distribution in the same eye lens was obtained by using the 1450-cm-1 CH₂ and CH₃ bending modes as an intrinsic marker for protein. It appears that the cholesterol is concentrated in the membranes of the eye lens fibers, whereas the protein is distributed more evenly. Second, we demonstrate that phenotyping of lymphocytes can be done by using the Raman scattering of (antibody-coated) polystyrene spheres. The lymphocyte population was also fluorescently labeled with anti-CD4-FITC to demonstrate that Raman and fluorescence labeling can be used simultaneously. Finally, we discuss the potential advantages and disadvantages of using Raman labels.