Tissue Clearing Composition for Morphology Preservation

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Solution Overview

Problem

Current methods for imaging complex tissues lack the ability to preserve cellular morphology and reporter fluorescence while allowing for multiplex phenotypic analysis, especially in three-dimensional structures, leading to limitations in understanding tissue organization and cellular interactions.

Innovation Solution

A tissue clearing composition comprising a compound of formula R1—C(X)—NR2R3, where R1 is alkyl, haloalkyl, hydroxyalkyl, amino, or alkylamino, and X is O or S, combined with a non-ionic density gradient medium, which permeates tissues to enhance transparency and compatibility with antibody-based immunolabeling, enabling high-resolution imaging and analysis.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Illumination intensity

If conventional tissue clearing methods are used to enhance tissue transparency, then imaging depth is improved, but cellular morphology and reporter fluorescence are lost

Engineering Contradiction:

Improveimaging depthVSAvoidcellular morphology preservation

Core Design Contradiction:

Illumination intensityVSReliability

Solution Approach 1:

The patent modifies the chemical composition parameters of clearing solutions by combining compounds of formula R1-C(X)-NR2R3 with non-ionic density gradient media, creating optimized formulations that achieve tissue transparency while preserving cellular structures and fluorescent signals through controlled refractive index matching and selective lipid removal

Inventive Principle:

Principle #35Parameter changes

2Loss of information

If tissue clearing is performed to enable three-dimensional imaging, then volumetric information is obtained, but multiplex phenotypic analysis capability is reduced

Engineering Contradiction:

Improvevolumetric informationVSAvoidmultiplex phenotypic analysis

Core Design Contradiction:

Loss of informationVSAdaptability or versatility

Solution Approach 1:

The patent employs composite clearing solutions containing multiple components (R1-C(X)-NR2R3 compounds combined with non-ionic density gradient media) that work synergistically to maintain tissue permeability for antibody penetration while preserving epitope integrity, enabling both 3D imaging and multiplex immunolabeling

Inventive Principle:

Principle #40Composite materials

3Illumination intensity

If existing clearing compositions are used to achieve tissue transparency, then optical clearing is improved, but compatibility with antibody-based immunolabeling is compromised

Engineering Contradiction:

Improvetissue transparencyVSAvoidcompatibility with immunolabeling

Core Design Contradiction:

Illumination intensityVSAdaptability or versatility

Solution Approach 1:

The non-ionic density gradient media in the patent act as intermediary substances that facilitate both optical clearing and antibody penetration, serving as a bridge between the clearing function and immunolabeling compatibility by providing a permissive chemical environment for antibody binding while maintaining tissue transparency

Inventive Principle:

Principle #24Intermediary (Mediator)

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 solution provides excellent tissue transparency, preserves cellular morphology and protein fluorescence, and allows for quantitative analysis of distinct cell populations, significantly improving imaging depth and maintaining tissue volume, thus enabling comprehensive understanding of cellular interactions and distributions.

Implementation Method 1

These techniques work by reducing light scattering in tissues through minimizing refractive index mismatches between the immersion medium and the various protein, aqueous, and lipid tissue constituents

Methodology Applied

Scientific EffectLight scattering reduction: Scattering

Implementation Method 2

reducing light scattering in tissues through minimizing refractive index mismatches between the immersion medium and the various protein, aqueous, and lipid tissue constituents

Methodology Applied

Scientific EffectRefractive index matching: Refraction

Data Source

PatentUS11480502B2Method and composition for optical clearing of tissues

Publication Date: 2022.10.25 THE GOVERNMENT OF THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SECRETARY DEPARTMENT OF HEALTH & HUMAN SERVICES

AI summary

Disclosed are compositions, methods, and kits for clearing tissue that preserve cellular morphology, reporter fluorescence, and epitope labeling which allow for quantitative phenotypic analysis of intact organs. The compositions include, for example, a compound of formula R1—C(X)—NR2R3, wherein R1 is alkyl, haloalkyl, hydroxyalkyl, amino, or alkylamino, X is O or S, and R2 and R3 are independently H, alkyl, or hydroxyalkyl, a salt thereof, or a combination thereof, and at least one non-ionic density gradient medium. Also disclosed are methods for clearing tissue comprising positioning a tissue in a tissue clearing composition and allowing a tissue clearing composition to permeate the tissue. Further disclosed are methods for visualizing tissue characteristics which involve fixing a tissue, staining the tissue, positioning the tissue in the tissue clearing composition and allowing the tissue clearing composition to permeate the tissue, and imaging the tissue utilizing a microscope or tissue scanning device.