PT  - JOURNAL ARTICLE
AU  - Wenjun Shao
AU  - Minzi Chang
AU  - Kevin Emmerich
AU  - Patrick O Kanold
AU  - Jeff S Mumm
AU  - Ji Yi
TI  - <em>Mesoscopic</em> oblique plane microscopy (Meso-OPM) - enabling large-scale 4D isotropic cellular resolution imaging with a diffractive light sheet
AID  - 10.1101/2022.03.29.486239
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.03.29.486239
4099  - http://biorxiv.org/content/early/2022/03/30/2022.03.29.486239.short
4100  - http://biorxiv.org/content/early/2022/03/30/2022.03.29.486239.full
AB  - Fundamental understanding of large-scale dynamic connectivity within a living organism requires volumetric imaging over a large field of view (FOV) at biologically relevant speed and resolution. However, most microscopy methods make trade-offs between FOV and resolution or imaging speed, making it challenging to observe highly dynamic processes at cellular resolution across mesoscopic scales (e.g., whole zebrafish larva). To overcome this limitation, we have developed mesoscopic oblique plane microscopy (Meso-OPM) with a diffractive light sheet. By augmenting the illumination angle of the light sheet with a transmission grating, the axial resolution was improved ~7-fold over existing methods and ~2-fold beyond the diffraction limitation of the primary objective lens. We demonstrated an unprecedented FOV up to 5.4 × 3.3 mm with resolution of 2.5× 3 × 6 μm, allowing volumetric imaging of cellular structures in an uncleared brain slice with a single scan. Applying Meso-OPM for in vivo imaging of zebrafish larvae, we report here the first whole fish volumetric recordings of neuronal activity at 2 Hz volume rate and the first example of whole fish volumetric recordings of blood flow dynamics at 5 Hz with isotropic cellular resolution.Competing Interest StatementThe authors have declared no competing interest.