Construction of an integrated Raman- and angular-scattering microscope

Abstract

We report on the construction of a multimodal microscope platform capable of gathering both elastically and inelastically scattered light from a 38 mum(2) region in both epi- and transillumination geometries. Simultaneous monitoring of elastic and inelastic scattering from a microscopic region allows noninvasive characterization of the chemistry and morphology of a living sample without the need for exogenous dyes or labels, thus allowing measurements to be made longitudinally in time on the same sample as it evolves naturally. A sample is illuminated either from above or below with a focused 785 nm TEM(00) mode laser beam, with elastic and inelastic scattering collected by two separate measurement arms. The measurements may be made either simultaneously, if identical illumination geometries are used, or sequentially, if the two modalities utilize opposing illumination paths. In the inelastic arm, Stokes-shifted light is dispersed by a spectrograph onto a charge-coupled device (CCD) array. In the elastic scattering collection arm, a relay system images the microscope's back aperture onto a CCD array. Postprocessing of the inelastic scattering to remove fluorescence signals yields high quality Raman spectra that report on the sample's chemical makeup. Comparison of the elastically scattered pupil images to generalized Lorenz-Mie theory yields estimated size distributions of scatterers within the sample.