METHOD AND SYSTEM FOR MULTI-VIEW EPISCOPIC SELECTIVE PLANE ILLUMINATION MICROSCOPE

METHOD AND SYSTEM FOR MULTI-VIEW EPISCOPIC SELECTIVE PLANE ILLUMINATION MICROSCOPE

Researchers at the Chan Zuckerberg Biohub have developed a single objective, multi-view light-sheet microscope with increased imaging depth and resolution.

Light-sheet microscopy, a type of selective plane illumination microscopy (SPIM), is an essential imaging technique that has been developed and used for volumetric imaging of biological samples using lower illumination intensities, resulting in reduced photobleaching and phototoxicity. However, a major limitation in SPIM is the requirement of multi-objective configurations that complicate sample mounting. Recent advances in single objective imaging configurations are limited in either resolution and sensitivity, field-of-view, or imaging speed.

Stage of Research

The inventors have developed methods and systems for a multi-view single objective light sheet microscope, capable of imaging large samples with large sample volumes. The authors describe several innovations: a novel custom tertiary objective and a new fast 3D scanning modality - light-sheet stabilized scanning (LS³) – which together extend the imaging volume without compromising resolution or speed. In addition, the inventors describe dual illumination and detection to enable multi-view imaging. Further, the inventors improve sample mounting ergonomics by converting the microscope from upright to inverted using remote focusing. The inventors validate the large imaging volume, speed, and resolution of their system by imaging whole zebrafish larvae and Drosophila egg chambers.

Applications

Volumetric, three-dimensional and time-resolved optical SPIM imaging of microscopic specimens with large sample volumes

Advantages

Large field-of-view and high resolution by means of a custom tertiary objective
Light-sheet stabilized stage scanning further increases imaging volume without compromising imaging speed
Dual light-sheet illumination and detection for multi-view imaging, enhancing volumetric coverage
Remote coverslip converts microscope from upright to inverted, enabling standard specimen mounting techniques, including multi-well plates, and on-stage sample manipulation or microfluidic device integration
Applicable to fluorescence imaging, bright field microscopy, quantitative phase measurements, polarization microscopy, and interferometric scattering microscopy

Publications

Yang B, Lange M, Millet-Sikking A, Solak AC, Kumar SV, Wang W, Kobayashi H, McCarroll MN, Whitehead LW, Fiolka RP, Kornberg TB, York AG, Royer LA. High-resolution, large imaging volume, and multi-view single objective light-sheet microscopy. 2020. bioRxiv. Doi: 10.1101/2020.09.22.309229

­METHOD AND SYSTEM FOR MULTI-VIEW EPISCOPIC SELECTIVE PLANE ILLUMINATION MICROSCOPE

METHOD AND SYSTEM FOR MULTI-VIEW EPISCOPIC SELECTIVE PLANE ILLUMINATION MICROSCOPE