Specimen Base for 3D Imaging of Biological Samples
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Solution Overview
Problem
Current tissue-clearing methods struggle to achieve isotropic high-resolution imaging throughout entire biological samples due to optical aberrations caused by refractive index mismatches and the limited image depth of high-numeric-aperture microscope objectives.
Innovation Solution
A specimen base and microscopic apparatus are designed to facilitate detachable coupling of top and bottom plates, enabling precise sectioning and 3D imaging. This system includes a microtome for sectioning and a microscope with an angle adjusting stage for optimal imaging, combined with tissue clearing techniques like Transparent Embedding Solvent System (TESOS) for enhanced transparency and mechanical strength.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If high-numeric-aperture microscope objectives are used to improve image resolution, then resolution is improved, but working distance is reduced which limits image depth
Solution Approach 1:
The imaging process is segmented into multiple steps: initial high-resolution imaging of the surface, followed by physical sectioning of the sample, and then imaging of subsequent layers. This allows using high-NA objectives for resolution while achieving deep imaging through sequential sectioning rather than requiring a single long working distance
Solution Approach 2:
The invention transitions from attempting to image deep in the Z-dimension with a single objective to creating multiple 2D sections that are then reconstructed into 3D. This dimensional transformation allows high-resolution imaging at each section while achieving depth through the stack of sections
2Ease of manufacture
If tissue clearing methods are used to improve transparency for deep imaging, then transparency is improved, but optical aberrations accumulate along long optical paths causing resolution deterioration
Solution Approach 1:
The sample is physically segmented into multiple thin sections rather than imaging through the entire thick sample. This segmentation reduces the optical path length through each imaging plane, preventing accumulation of optical aberrations while still achieving deep overall imaging through section stacking
Solution Approach 2:
Tissue clearing and embedding are performed as preliminary actions before sectioning and imaging. This prepares the sample for optimal optical properties in each thin section, ensuring maximum transparency and minimum aberration accumulation along the shortened optical path of each section
3Adaptability or versatility
If detachable coupling of top and bottom plates is implemented to enable precise sectioning and 3D imaging, then imaging flexibility is improved, but device complexity increases
Solution Approach 1:
The detachable plate system is designed with universal features: standardized coupling mechanisms, interchangeable plates for different sample types, and compatibility with multiple imaging modalities. This multi-functionality justifies the added complexity by enabling diverse applications from sectioning to 3D reconstruction
Solution Approach 2:
The detachable plates serve as intermediaries between the sectioning process and the imaging process. They provide a standardized interface that couples the mechanical sectioning capability with the optical imaging capability, allowing precise positioning and stable mounting while maintaining flexibility
Data Source
AI summary
A specimen base, a specimen sectioning device, a microscopic apparatus and a method for 3D imaging of biological specimen. The specimen base comprising a top plate (1) and a bottom plate (2) that are detachably coupled together; the top plate (1) comprises a first plane (11) and a second plane (12) opposite each other, the first plane (11) is used for fixing specimen, and the second plane (12) is coupled with a third plane (21) of the bottom plate (2); a top plate fixing part (13) is provided on the second plane (12) and a bottom plate fixing part (22) is provided at the corresponding position on the third plane (21); the top plate (1) couples with the bottom plate (2) by means of matching the top plate fixing part (13) with the bottom plate fixing part (22); a through-hole (23) is provided in the bottom plate (2).