Spatially resolved micro-absorption spectroscopy with a broadband source and confocal detection

We present a novel approach to measure optical absorption spectra with spatial resolution at the micron scale. The setup combines a continuous white light excitation beam in transmission geometry with a confocal microscope. The spatial resolution is found to be better than 1.4 μm in the lateral and 3.6 μm in the axial direction. Employing multichannel detection the absorption spectrum of hemoglobin in a single red blood cell is measured on the timescale of seconds. Through measurements of the transmitted intensity in solutions in nanoliter quantities we establish that the absorbance varies linearly with concentration. Our setup enables the investigation of spatial variations in the optical density of small samples on the micron scale and can be applied to the study of biological assemblies at the single cell level, in optical diagnostics, and in micro-fluidics.