Performance tradeoffs for single- and dual-objective open-top light-sheet microscope designs: a simulation-based analysis
Abstract
Light-sheet microscopy (LSM) has emerged as a powerful tool for high-speed volumetric imaging of live model organisms and large optically cleared specimens. When designing cleared-tissue LSM systems with certain desired imaging specifications (e.g. resolution, contrast, and working distance), various design parameters must be taken into consideration. In order to elucidate some of the key design tradeoffs for LSM systems, we present a diffraction-based analysis of single- and dual-objective LSM configurations using simulations of LSM point spread functions. We assume Gaussian illumination is utilized. Specifically, we analyze the effects of the illumination and collection numerical aperture (NA), as well as their crossing angle, on spatial resolution and contrast. Assuming an open-top light-sheet (OTLS) architecture, we constrain these parameters based on fundamental geometric considerations as well as those imposed by currently available microscope objectives. In addition to revealing the performance tradeoffs of various single- and dual-objective LSM configurations, our analysis showcases the potential advantages of a novel, non-orthogonal dual-objective (NODO) architecture, especially for moderate-resolution imaging applications (collection NA of 0.5 to 0.8).
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Jul 24, 2020
- Source ID
- 10.1364/boe.397052
Entities
People
- Adam K Glaser
- Jonathan T C Liu
- Kevin W Bishop
Organizations
- National Institutes of Health
- National Science Foundation
- United States Department of Defense