Light Sheet Light Field Microscopy

in collaboration with Diego Castro, Manu Forero-Shelton and the Biophysics Laboratory at Universidad de los Andes

Many biological processes of interest exhibit rapid dynamics, such as calcium signaling in the brain or the beating of the heart. In embryos, these processes can be observed under the microscope. However, a microscope capable of registering these dynamics must capture large fields of view at high spatial and temporal resolution.

Light sheet microscopy allows to register high spatial-resolution tomographic views of live samples. Based on optical sectioning, only the portion of the sample which is at focus is illuminated at any given time, enhancing contrast and resolution. This comes at the cost of slow, sequential acquisition, having to illuminate and photograph each plane within a volume to render a full view.

Figure1. A light field capture of an ink spill. An artistic interpretation.

Light field microscopy, on the other hand, permits producing full volumetric reconstructions after a single shot. By capturing information on the origin and direction of light rays, light fields are suited to synthetically refocus at any given plane with post-processing. To gain in frame rate, however, one must sacrifice spatial resolution.

By combining optical sectioning with the possibility to reconstruct full volumes in-focus after a single shot, Light Sheet Light Field Microscopy offers high spatial and high temporal resolution. In collaboration with the Biophysics Laboratory at Universidad de los Andes, we have developed a Light Sheet Light Field Microscopy and its ancillary software. We have succeeded in using light sheet and light field as complementary techniques to modulate spatial and temporal resolution of the microscope without modifying the hardware.

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