Confocal LiDAR for remote high-resolution imaging of auto-fluorescence in aquatic media

Spatially resolved in situ monitoring of plankton can provide insights on the impacts of climate change on aquatic ecosystems due to their vital role in the biological carbon pump. However, high-resolution underwater imaging is technically complex and restricted to small close-range volumes with current techniques. Here, we report a novel inelastic scanning confocal light detection and ranging (LiDAR) system for remote underwater volumetric imaging of fluorescent objects. A continuous wave excitation beam is combined with a pinhole in a conjugated detection plane to reject out-of-focus scattering and accomplish near-diffraction limited probe volumes. The combination of bi-directional scanning with remote focusing enables the acquisition of three-dimensional data. We experimentally determine the point spread and axial weighting functions, and demonstrate selective volumetric imaging of obstructed layers through spatial filtering. Finally, we spatially resolve in vivo autofluorescence from sub-millimeter Acocyclops royi copepods to demonstrate the applicability of our novel instrument in non-intrusive morphological and spectroscopic studies of aquatic fauna. The proposed system constitutes a unique tool e.g. for profiling chlorophyll distributions and for quantitative studies of zooplankton with reduced interference from intervening scatterers in the water column that degrade the the performance of conventional imaging systems currently in place.