PT  - JOURNAL ARTICLE
AU  - Bin Yang
AU  - Alfred Millett-Sikking
AU  - Merlin Lange
AU  - Ahmet Can Solak
AU  - Hirofumi Kobayashi
AU  - Andrew York
AU  - Loic A. Royer
TI  - High-Resolution, Large Field-of-View, and Multi-View Single Objective Light-Sheet Microscopy
AID  - 10.1101/2020.09.22.309229
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.09.22.309229
4099  - http://biorxiv.org/content/early/2020/09/24/2020.09.22.309229.short
4100  - http://biorxiv.org/content/early/2020/09/24/2020.09.22.309229.full
AB  - Light-sheet microscopy has become the preferred method for long-term imaging of large living samples because of its low photo-invasiveness and good optical sectioning capabilities. Unfortunately, refraction and scattering often pose obstacles to light-sheet propagation and limit imaging depth. This is typically addressed by imaging multiple complementary views to obtain high and uniform image quality throughout the sample. However, multi-view imaging often requires complex multi-objective configurations that complicate sample mounting, or sample rotation that decreases imaging speed. Recent developments in single-objective light-sheet microscopy have shown that it is possible to achieve high spatio-temporal resolution with a single objective for both illumination and detection. Here we describe a single-objective light-sheet microscope that achieves: (i) high-resolution and large field-of-view imaging via a custom remote focusing objective; (ii) simpler design and ergonomics by remote placement of coverslips; (iii) fast volumetric imaging by means of light-sheet stabilised stage scanning – a novel scanning modality that extends the imaging volume without compromising imaging speed nor quality; (iv) multi-view imaging by means of dual orthogonal light-sheet illumination. Finally, we demonstrate the speed, field of view and resolution of our novel instrument by imaging zebrafish tail development.Competing Interest StatementThe authors have declared no competing interest.