Hyperspectral Oblique Plane Microscopy Enables Spontaneous, Label-Free Imaging of Biological Dynamic Processes in Live Animals

Abstract

Spontaneous Raman imaging has emerged as powerful label-free technique for investigating the molecular composition of biological specimens. Although Raman imaging can facilitate understanding of complex biological phenomena in vivo, current imaging modalities are limited in speed and sample compatibility. Here, we introduce a single-objective light-sheet microscope, named{lambda} -OPM, which records Raman images on a timescale of minutes to milliseconds. To demonstrate its function, we use{lambda} -OPM to map and identify micro-plastic particles based on their Raman spectral characteristics. In live zebrafish embryos, we show that{lambda} -OPM can capture wound dynamics at five-minute intervals, revealing rapid changes in cellular and extracellular matrix composition in the wounded region. Finally, we use{lambda} -OPM to obtain Raman scattering maps of a zebrafish embryos beating heart at an effective 28 frames per second, recording compositional changes at different points in the cardiac cycle.