A line-scanning hyperspectral Raman microscope based on Bessel beam illumination

Carbohydrate metabolism within the photosynthetic processes of plants plays a major role in plant performance. Current techniques for live imaging the structural characteristics of plants are often destructive or lack spatially explicit information. Raman spectroscopy is a nondestructive, label-free imaging technique which, in combination with Bessel beam light sheet microscopy, offers a way to rapidly measure the abundance and flux of molecules as they travel through the three dimensions of living plants. Here we describe the development of a Hyperspectral Raman Light Sheet Microscope as an alternative to traditional confocal point-scanning imaging systems. We cover our methods for creating a self-reconstructing Bessel beam to excite scattering in the sample, as well as modifications made to the conventional slit-spectrometer setup for capturing low intensity Raman scattered infrared light. We demonstrate the system with various test samples including Fluorinated Ethylene Propylene and Acetaminophen, and then discuss future applications in biological materials.