Multiscale Light Field Microscopy Platform for Multi-purpose Dynamic Volumetric Bioimaging

Light field microscopy (LFM) has emerged in recent years as a unique solution for fast, scan-free volumetric imaging of dynamic biological samples. This is achieved by using a microlens array in the detection path to record both the lateral and angular information of the light fields coming from the sample, capturing a 3-dimensional (3D) volume in a single 2-dimensional (2D) snapshot. In post-acquisition, the 3D sample volume is computationally reconstructed from the recorded 2D images, thus enabling unprecedented 3D capture speed, not limited by the typical constraint of physically scanning the focal plane over the sample volume. Up to date, most published LFM imaging setups have been specialized single-purpose platforms, optimized for a narrow performance window in field of view and resolution, thus hampering widespread adoption of LFM for biomedical research. Here, we present a versatile LFM platform for fast 3D imaging across multiple scales, enabling applications from cell to system-level biology on the same imaging setup. Our multiscale LFM is built as an add-on module to a conventional commercially available wide field microscope, and the various imaging applications, with different ranges of field of view and resolution, are achieved by simply switching between the standard microscope objectives available on the wide field microscope. Additionally, we provide an open-source end-to-end software package for calculating the system performance parameters, processing the experimentally measured point spread function, and light field 3D image reconstruction. We demonstrate the performance of our multiscale LFM platform through imaging the whole-brain activity map of seizures in larval zebrafish, calcium dynamics in ex vivo mouse pancreatic islets, and subcellular protein dynamics in cultured cells.