Whole-Body Tissue Clearing for Single-Cell Biodistribution Imaging
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Solution Overview
Problem
Current methods for visualizing tumor metastases and biopharmaceutical drug distribution in whole animals lack the ability to provide unbiased, high-resolution imaging at the single-cell level without dissection, and existing tissue clearing techniques are limited by autofluorescence and require specialized equipment.
Innovation Solution
A method combining whole-body labeling with nanobodies and organic solvent-based clearing (LuCiD) to create optically transparent tissues, allowing visualization of single cells and molecules in intact animals using commonly available microscopes, and enabling detection of tumor metastases and drug distribution at the cellular level.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Length of stationary object
If tissue clearing methods are used to render tissues transparent, then imaging depth and penetration are improved, but autofluorescence increases and measurement precision deteriorates
Solution Approach 1:
The patent shifts the imaging wavelength from visible to near-infrared range (700-1000 nm), changing the optical parameters to avoid autofluorescence while maintaining deep tissue penetration. This parameter change allows imaging through cleared tissues without the background interference that plagues visible light microscopy.
Solution Approach 2:
The patent uses light-sheet fluorescence microscopy to create optical copies and 3D reconstructions of entire organs and tissues. This allows visualization of deep tissue structures by capturing multiple planes and reconstructing them computationally, bypassing the need for physical sectioning while maintaining measurement precision.
2Area of stationary object
If whole-body labeling is performed to visualize target molecules throughout the animal, then comprehensive coverage is improved, but detection precision deteriorates due to signal dilution
Solution Approach 1:
The patent extracts and amplifies specific fluorescent signals from target molecules using near-infrared imaging, separating the signal of interest from the background. By focusing detection on specific wavelengths and using signal amplification techniques, the method maintains high detection precision even when visualizing targets throughout the entire body.
Solution Approach 2:
The patent employs a universal labeling approach using near-infrared fluorophores that can label various target molecules (proteins, cells, structures) throughout different organs and tissues. This multi-functional labeling system maintains detection precision across diverse biological targets while providing comprehensive whole-body coverage.
3Measurement precision
If dissection is performed to analyze specific organs, then measurement precision is improved for localized regions, but loss of information increases regarding overall distribution
Solution Approach 1:
The patent transitions from 2D tissue section analysis to 3D whole-organ imaging using light-sheet fluorescence microscopy. By imaging entire organs in three dimensions without physical sectioning, the method maintains high localization precision for specific structures while simultaneously preserving information about their spatial distribution throughout the entire organism.
Solution Approach 2:
The patent uses computational segmentation to divide the 3D imaging data into distinct anatomical regions and structures of interest. This allows precise measurement and analysis of specific organs or tissues while maintaining the context of their overall distribution and relationships within the complete organism.
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 unbiased, high-throughput mapping of tumor micrometastases and precise assessment of biopharmaceutical drug biodistribution in whole animals, reducing the need for dissection and specialized equipment, and providing cost-effective, time-efficient insights into cancer therapy efficacy.
Implementation Method 1
Clearing the fixed animal tissue labeled with the fluorochrome-containing labeling agent with a clearing solution comprising an organic solvent
Implementation Method 2
Labeling a target molecule in the fixed animal tissue with a labeling solution comprising a fluorochrome-containing labeling agent capable of binding to the target molecule
Data Source
AI summary
The present invention relates to methods for preparing an animal tissue for fluorescence microscopy, to an animal tissue obtainable by said methods, to methods for analyzing said animal tissues, and to methods for the detection of metastases, for analyzing the biodistribution of a biopharmaceutical drug, and for analyzing the biodistribution of nanoparticles. The methods for preparing an animal tissue according to the present invention encompass whole-body labeling, clearing and imaging methods. The methods of the invention are advantageous in that they, for instance, allow the visualization of single cells within mammalian tissues including pig and human brains, tumor metastases at the single cell level and of the distribution of biopharmaceutical drugs (e.g. the distribution of cancer-targeting therapeutic antibodies in whole animals such as intact mice) at single cell level in whole mouse.