Open-Top Light-Sheet Microscopy With Non-Orthogonal Objectives
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Solution Overview
Problem
Existing microscopes with orthogonal illumination and collection objectives face limitations in resolution and imaging depth due to high NA objectives' shorter working distances and strict refractive index matching requirements, which constrain specimen thickness and impose lateral constraints.
Innovation Solution
A non-orthogonal arrangement of illumination and collection objectives, allowing for higher NA collection objectives and reduced index matching requirements, with dual collection paths for high and low magnification modes, and adjustable illumination optics for enhanced imaging.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If orthogonal illumination and collection objectives are used with high NA, then resolution is improved, but working distance decreases and lateral constraints are imposed
Solution Approach 1:
The patent employs a non-orthogonal arrangement where the illumination objective and collection objective are positioned at an angled configuration rather than perpendicular to each other. This asymmetric geometry allows the collection objective to achieve high NA for improved resolution while maintaining adequate working distance by orienting its optical axis at an angle to the illumination path, thereby resolving the contradiction between resolution and working distance
Solution Approach 2:
The patent introduces an additional angular dimension to the optical configuration by tilting the illumination objective relative to the collection objective. This dimensional change allows the system to decouple the constraints of high NA collection from lateral working distance limitations, enabling high-resolution imaging without the severe lateral constraints imposed by orthogonal high-NA configurations
2Measurement precision
If orthogonal illumination and collection objectives are used with high NA, then resolution is improved, but refractive index matching requirements become stricter
Solution Approach 1:
The non-orthogonal arrangement creates an asymmetric optical path where light travels through different media interfaces at varied angles. This asymmetry reduces the sensitivity to refractive index mismatches compared to orthogonal high-NA configurations, as the angled illumination path is less susceptible to refraction errors at interface boundaries, thereby relaxing the strict index matching requirements while maintaining high resolution
3Ease of operation
If open-top configuration is used, then convenience for imaging tissue specimens is improved, but resolution and imaging depth are limited
Solution Approach 1:
The patent implements adjustable and reconfigurable optical components that allow dynamic optimization of the imaging geometry. The illumination and collection objectives can be positioned and angled to achieve optimal resolution for different specimen types and depths while preserving the open-top configuration's operational convenience, enabling the system to adapt between convenience and resolution requirements
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 lateral constraints and relaxed refractive index matching, facilitating both screening and detailed analysis of samples.
Implementation Method 1
The illumination objective is configured to direct an illumination light sheet along an illumination axis into a sample
Implementation Method 2
The collection objective is configured to receive light from an imaging plane of the sample along a collection axis
Implementation Method 3
The non-orthogonal arrangement of illumination and collection objectives... reduced index matching requirements
Data Source
Figure 1
Figure 2A~2B
Figure 2C~2D
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.