Tissue Specimen Analysis via Aqueous Clearing and 3D Imaging
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Solution Overview
Problem
Conventional histological examination methods face challenges in accurately diagnosing diseases due to tissue distortion and loss of morphological information during sample preparation, particularly from dehydration and slicing, which affects the representativeness and quality of sample images.
Innovation Solution
A method involving tissue treatment with an aqueous clearing agent and fluorescent probes to generate 3D images, which are then matched with 2D images to extract corresponding slices and determine pathological scores for accurate disease diagnosis, using a system that includes a processor to facilitate these operations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Manufacturing precision
If conventional histological examination methods are used (fixation, embedding, slicing, staining), then tissue sections can be prepared for visual analysis, but tissue distortion and loss of morphological information occur due to dehydration and slicing
Solution Approach 1:
The patent changes the physical and chemical parameters of the tissue processing method by using aqueous clearing agents instead of traditional organic solvents, and by performing imaging on cleared whole-mount specimens before slicing. This preserves the native 3D tissue architecture and morphological information while still enabling detailed analysis through subsequent 2D sectioning of the already-imaged specimen.
Solution Approach 2:
The patent transitions from traditional 2D analysis of sliced sections to 3D imaging of whole-mount cleared tissue specimens. By capturing images in the third dimension (depth) before slicing, the method preserves complete morphological information that would otherwise be lost in sequential 2D sections, allowing reconstruction and analysis of the full tissue architecture.
2Loss of information
If 3D imaging is performed on conventional sliced tissue sections, then virtual 3D reconstruction can be achieved, but gaps and artifacts from tissue discontinuity and preparation remain
Solution Approach 1:
The patent performs 3D imaging of the entire tissue specimen in its native, intact state before any slicing or sectioning occurs. This preliminary action captures complete morphological information and eliminates gaps between sections, as the imaging is performed on the continuous, unsliced tissue. Subsequent slicing is done after the 3D data has already been acquired, ensuring no information is lost.
3Illumination intensity
If traditional clearing agents are used, then tissue transparency can be achieved, but tissue shrinkage and deformation occur
Solution Approach 1:
The patent uses aqueous clearing agents with specific refractive indices matched to tissue components, rather than traditional organic solvents. This parameter change in the clearing medium composition achieves tissue transparency without causing shrinkage or deformation, as the aqueous environment better preserves native tissue morphology while still enabling optical clearing for imaging.
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
This approach enhances diagnostic accuracy by preserving tissue structural integrity, reducing artifacts, and providing more reliable morphological information, leading to improved disease detection and treatment decisions.
Implementation Method 1
The aqueous clearing agent has a refractive index of 1.33-1.55... causes a tissue specimen with a thickness of at least 200 μm to become sufficiently transparent
Implementation Method 2
treating a tissue specimen from a subject with an aqueous clearing agent and with at least two fluorescent probes for labeling cell membrane and cell nuclei
Data Source
AI summary
Provided is a method for analyzing a tissue specimen, including treating a tissue specimen with an aqueous clearing agent and with at least two fluorescent probes to obtain a cleared and labeled tissue specimen; imaging the cleared and labeled tissue specimen to generate a three-dimensional (3D) image of the tissue specimen; preparing a stained tissue section from the cleared and labeled tissue specimen; capturing a reference two-dimensional (2D) image of the stained tissue section; matching the reference 2D image with the 3D image to extract from the 3D image a series of 2D image slices including a corresponding 2D image slice that corresponds to the reference 2D image; and determining at least one pathological score for each of the series of 2D image slices and reporting the presence or absence and the extent of the disease based on the pathological scores. The method can improve the accuracy of histopathologic diagnosis.