PT  - JOURNAL ARTICLE
AU  - Kevin Keomanee-Dizon
AU  - Scott E. Fraser
AU  - Thai V. Truong
TI  - A versatile twin-microscope system for light-sheet imaging
AID  - 10.1101/801688
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 801688
4099  - http://biorxiv.org/content/early/2019/10/12/801688.short
4100  - http://biorxiv.org/content/early/2019/10/12/801688.full
AB  - Recent developments in light-sheet microscopy techniques has led to its cutting-edge application across a range of fields, from developmental biology to behavioral neuroscience. Its low photodamage propensity allows biological dynamics to be imaged for a duration of several hours up to days; however, a tradeoff exists between such extended imaging sessions and imaging throughput, which in practice directly limits analysis workflows and the resulting time to biological discovery. Here, we present an economical instrument that can image multiple sample types simultaneously. Unlike any existing light-sheet microscope, our instrument, the flex-SPIM, shares both an ultrafast laser and a bank of continuous-wave lasers between two independently controlled light-sheet microscope-twins. Harnessing the power of modern lasers yields a substantial reduction in the cost of the light source over standard dual systems, and a doubling in imaging throughput relative to a single system. Each microscope-twin (i) operates in both one-photon and two-photon excitation modes, (ii) delivers one to three light-sheets via a trio of orthogonal excitation arms, (iii) provides sub-micron to micron imaging resolution, (iv) is multicolor compatible, and (v) permits upright and/or inverted light-sheet detection. We offer a detailed description of the flex-SPIM design to aid instrument builders who wish to construct and use a similar system. We then demonstrate the instrument’s versatility for biological investigation by performing fast imaging of the beating heart in an intact zebrafish embryo, deep imaging of thick patient-derived tumor organoids, and gentle whole-brain imaging of neural activity in behaving larval zebrafish.