Light Sheet Microscope Overview Illumination via Lateral Detection
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Solution Overview
Problem
Light sheet microscopy with a 45° SPIM arrangement faces challenges in capturing high-contrast overview images due to the inability to integrate transmitted light illumination, especially when imaging through liquid, leading to issues like meniscus formation and aberrations in multiwell plates.
Innovation Solution
A microscope design with an illumination lens and detection lens aligned to intersect at the sample plane, allowing for oblique overview illumination without apertures, and utilizing diffuse or polarized illumination to enhance contrast, which can be achieved without immersing the illumination optics in liquid, thus avoiding lens effects and aberrations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If a wide-field lens is positioned below the sample to capture overview images, then overview imaging is enabled, but the lens must look through several millimeters of liquid causing meniscus formation and lens effects that degrade image quality
Solution Approach 1:
The patent positions the wide-field lens laterally adjacent to the sample rather than below it, changing the spatial dimension of observation. This allows the lens to view the sample edge-on through the side wall of the sample holder, eliminating the need to look through the liquid column and avoiding meniscus-related optical distortions.
Solution Approach 2:
The patent creates an optical copy of the sample by imaging it through the side wall of the sample holder using the wide-field lens. This side-view optical copy provides overview information without requiring the lens to be positioned in the problematic below-sample location, thus avoiding liquid-related optical artifacts.
2Measurement precision
If the illumination and detection lenses are positioned close together in a 45° configuration, then high axial resolution is achieved, but transmitted light illumination cannot be implemented for overview imaging
Solution Approach 1:
The patent separates the illumination and detection functions into distinct optical paths. The light sheet illumination system maintains the 45° configuration for high-resolution imaging, while a separate wide-field illumination system provides transmitted light illumination for overview images, allowing both functions to operate independently without interference.
Solution Approach 2:
The detection lens serves dual purposes: it detects light from the light sheet for high-resolution imaging and simultaneously detects transmitted light from the wide-field illumination for overview images. This multi-functionality eliminates the need for separate detection optics while maintaining both imaging capabilities.
3Adaptability or versatility
If additional optics are added to enable transmitted light illumination, then overview imaging capability is improved, but the microscope structure becomes more complex
Solution Approach 1:
The detection lens is designed to perform multiple functions: detecting light from the light sheet for high-resolution imaging and detecting transmitted light from wide-field illumination for overview images. This multi-functionality reduces the need for additional separate detection optics, thereby limiting the increase in system complexity.
Solution Approach 2:
The patent combines the overview imaging capability with the existing detection optics by positioning the wide-field lens laterally and utilizing the detection lens's ability to capture transmitted light. This merging approach integrates multiple imaging modalities into a unified optical system rather than adding completely separate subsystems.
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-contrast overview images to be captured efficiently, reducing the need for additional optics and simplifying the microscope structure, while maintaining high resolution and minimizing contamination risks in high-throughput screening.
Implementation Method 1
an illumination lens (2) designed to generate a light sheet (6) with a first optical axis (A1), the light sheet (6) being generated or capable of being generated at least partially in a sample plane (4)
Implementation Method 2
a detection lens (3) designed to detect light coming from the sample plane (4) and having a second optical axis (A2)
Implementation Method 3
an overview illumination device designed for wide-field illumination of the sample plane (4) and having illumination optics with a third optical axis (A3)
Data Source
Figure 1a~1b
Figure 2a~2b
Figure 3a~3b
AI summary
The invention relates to a microscope (1) comprising an illumination objective (2) designed to generate a light sheet (6) with a first optical axis (A1) and a detection objective (3) designed to detect light emanating from the sample plane (4) with a second optical axis (A2). The illumination objective (2) and the detection objective (3) are aligned with each other and with the sample plane (4) such that the first optical axis (A1) and the second optical axis (A2) intersect in the sample plane (4) and form a substantially right angle with each other. The optical axes (A1, A2) each form a non-zero angle with a reference axis (B) directed orthogonally to the sample plane (4). Furthermore, an overview illumination device designed for wide-field illumination of the sample plane (4) is provided, comprising an illumination optic (9) with a third optical axis (A3).A key feature is that the detection objective (3) is designed and configured to detect both light from the light sheet (6) and light from the illumination optics (9). The invention further relates to a method for operating a light sheet microscope.