Diffractive oblique plane microscopy

Imaging of neuronal activity with fluorescent indicators is an important technique in neuroscience. However, it remains challenging to record volumetric image data at fast frame rates and good resolution. One promising technique to achieve this goal is light sheet microscopy (LSM), but the right angle configuration of the excitation and imaging system limits its application. Oblique plane microscopy (OPM), a variant of LSM, circumvents this limitation by exciting oblique planes and detecting the image through the same microscope objective lens. So far, these techniques have relied on the use of high numerical aperture (NA) detection objective lenses, which limits their field of view. Here we present an OPM technique that allows for the use of low NA objective lenses by redirecting the light with the help of a diffraction grating. The microscope maintains a micrometer-scale lateral resolution over a large addressable imaging volume of 3.3 x 3.0 x 1.0 mm(3). We demonstrate its practicality by imaging the whole brain of larval and juvenile zebrafish. (C) 2019 Optical Society of America under the terms of the OSA Open Access Publishing Agreement