Open-Top Light-Sheet Microscopy With Non-Orthogonal Objective Layout
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Solution Overview
Problem
Existing open-top light-sheet (OTLS) microscopes face limitations in resolution and imaging depth due to orthogonal geometry between illumination and collection objectives, which restricts high-resolution imaging of thicker specimens and imposes strict refractive index matching requirements.
Innovation Solution
A non-orthogonal arrangement of illumination and collection objectives, allowing for higher numerical aperture collection objectives and reduced index matching constraints, with optional dual collection paths for high and low magnification modes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If orthogonal geometry is used between illumination and collection objectives, then the system maintains simple alignment, but imaging depth and resolution are limited
Solution Approach 1:
The patent employs asymmetric optical geometry where the collection objective is positioned at a non-orthogonal angle (e.g., 45 degrees) relative to the illumination light sheet, breaking the conventional orthogonal symmetry. This asymmetric arrangement enables the collection objective to capture light from deeper imaging planes while maintaining high numerical aperture, thereby improving imaging depth and resolution without requiring complex adaptive alignment mechanisms
2Manufacturing precision
If high numerical aperture collection objectives are used, then imaging resolution improves, but refractive index matching requirements become stricter
Solution Approach 1:
The patent introduces an immersion medium as an optical intermediary between the collection objective and the sample. This immersion medium has a refractive index specifically matched to both the high NA collection objective and the tissue specimen, serving as a refractive index bridge. This intermediary layer enables the use of high numerical aperture objectives while maintaining relaxed refractive index matching tolerances, thereby improving resolution without compromising system reliability
3Ease of operation
If open-top configuration is used, then sample accessibility improves, but lateral constraints are imposed on imaging
Solution Approach 1:
The patent transitions from conventional lateral (x-y plane) imaging constraints to three-dimensional volumetric imaging capability. By positioning the collection objective at a non-orthogonal angle to the light sheet, the system enables optical sectioning through the z-dimension, allowing imaging of thick tissue volumes without lateral constraints. This dimensional transition maintains the open-top configuration's sample accessibility while adding versatility in imaging depth and volume
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
Enables high-resolution imaging of thicker specimens with reduced refractive index constraints and increased imaging depth, facilitating both screening and detailed analysis without lateral constraints.
Implementation Method 1
The illumination objective directs an illumination light sheet along an illumination axis into a sample
Implementation Method 2
The collection objective receive light from an imaging plane of the sample along a collection axis
Data Source
AI summary
Apparatuses, systems, and methods for an open-top light-sheet (OTLS) microscope which includes an illumination objective and a collection objective which have optical axes which are non-orthogonal to each other. The optical axis of the collection objective may be orthogonal to a plane of the sample holder. The illumination and collection objective may be located below the sample holder. The OTLS microscope may optionally include a second collection objective which has an optical axis orthogonal to the optical axis of the illumination objective. The illumination objective may be an air objective, and the collection objective may be an immersion objective.