Adaptive Optical Correction for Light Sheet Microscopy
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Solution Overview
Problem
Light sheet microscopy faces challenges with imaging errors due to the oblique passage of light through cover glasses of varying thicknesses and angles, which are not effectively addressed by existing correction methods, leading to compromised image quality and compatibility issues with standard sample holders.
Innovation Solution
Incorporating adaptive optical correction elements, such as wavefront manipulators with free-form surfaces, into the detection and illumination objectives to correct aberrations caused by varying cover glass thicknesses and angles, allowing for improved image quality and compatibility with standard sample vessels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If adaptive optical correction elements are incorporated into detection and illumination objectives, then image quality and correction effectiveness are improved, but device complexity increases
Solution Approach 1:
The patent introduces adaptive optical correction elements as intermediary components between the separating layer system and the detection/illumination objectives. These correction elements act as mediators that compensate for imaging errors caused by the separating layer, thereby improving image quality without requiring fundamental changes to the entire microscopy system.
Solution Approach 2:
The correction elements utilize adjustable optical parameters such as wavefront manipulation and free-form surface geometries to dynamically compensate for variations in cover glass thickness and angle. By changing these optical parameters adaptively, the system maintains high image quality across different sample holder configurations.
2Ease of operation
If standard sample holders are used with oblique illumination angles, then compatibility and ease of operation are improved, but imaging errors due to varying cover glass thickness and angles increase
Solution Approach 1:
The patent employs asymmetric free-form surfaces in the correction elements that are specifically designed to compensate for the asymmetric oblique passage of light through the separating layer system. This asymmetric design allows the correction elements to effectively address imaging errors caused by varying cover glass thicknesses and angles while maintaining compatibility with standard sample holders.
3Ease of manufacture
If correction methods are simplified for ease of manufacture, then manufacturing cost and complexity are reduced, but effectiveness in correcting aberrations caused by varying cover glass parameters decreases
Solution Approach 1:
The correction elements incorporate dynamic adjustment capabilities that allow them to adapt to varying cover glass thicknesses and angles during operation. This dynamic behavior enables a single correction element design to handle a range of parameters effectively, maintaining high correction effectiveness without requiring multiple specialized components for different scenarios.
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
The adaptive optical correction elements effectively reduce imaging errors, enabling diffraction-limited imaging over a range of cover glass thicknesses and angles, and facilitate the use of standard sample holders, enhancing the versatility and accuracy of light sheet microscopy.
Implementation Method 1
both the first illumination correction element and the first detection correction element are designed as wavefront manipulators
Implementation Method 2
aberrations that occur due to the oblique passage of light to be detected or of light for illuminating the sample through interfaces of the separating layer system
Data Source
Figure 1
Figure 2~3
Figure 4a
AI summary
The invention relates to an arrangement for light sheet microscopy. Said arrangement comprises an illumination optical unit having an illumination objective (5) for illuminating a sample (3), which is situated on a sample carrier in a medium (2), with a light sheet via an illumination beam path. The optical axis (6) of the illumination objective and the light sheet lie in a plane which forms an illumination angle (β) which is different from zero with the normal to a planar reference surface (4) with respect to which the sample carrier is aligned. The arrangement furthermore comprises a detection optical unit having a detection objective (7) in a detection beam path, the optical axis (8) of which forms a detection angle (δ) which is different from zero with the normal to the reference surface (4). The arrangement finally comprises a separating layer system having at least one layer composed of a predefined material having a predefined thickness which separates the medium (2) from the illumination objective (5) and the detection objective (7), wherein the separating layer system is in contact with the medium (2) with a base surface aligned parallel to the reference surface (4), at least in the region accessible for the illumination objective (5) and the detection objective (7) for illumination and detection, respectively. In such an arrangement, the detection objective (7) comprises a first adaptive optical detection correction element, which is arranged in the beam path or can be introduced into the latter. Alternatively or additionally, the illumination objective (5) comprises a first adaptive optical illumination correction element, which is arranged in the beam path or can be introduced into the latter. The two correction elements make it possible to reduce aberrations which arise on account of the oblique passage of light to be detected and/or of light for illuminating the sample (3) through interfaces of the separating layer system, for a predefined range of detection angles (δ) and/or of illumination angles (β) and/or for a predefined range of the thickness of the at least one layer of the separating layer system.