Expanded Biological Sample Imaging for Cellular Ultrastructure
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Solution Overview
Problem
Fluorescence microscopy is limited in revealing cellular ultrastructures due to insufficient optical resolution and the inability to label the entire sample at high density, while electron microscopy requires extensive data acquisition time for three-dimensional imaging.
Innovation Solution
A method involving physical expansion of biological samples by a factor of two or more, combined with bulk labeling using reagents to introduce contrast, allowing for high-density labeling and resolution of fine structures without specific labeling or electron microscopy.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If fluorescence microscopy is used to image proteins, then high contrast and high precision labeling are achieved, but the optical resolution is insufficient to reveal fine structures (ultrastructure) of the cell
Solution Approach 1:
The patent applies physical expansion to transform the sample from its native scale to an expanded scale (4-10x or more), effectively adding a dimensional transformation that enlarges fine structures to be resolvable by light microscopy. This dimensional change allows structures previously below the diffraction limit to become visible without changing the microscopy technique itself.
Solution Approach 2:
The patent changes the physical parameters of the sample by expanding its size while preserving the relative positions of cellular structures. This parameter change (size expansion) transforms unresolvable fine structures into resolvable ones, while the labeling precision remains high because the expansion maintains spatial relationships.
2Quantity of substance
If the rest of the sample is labeled to reveal cellular context, then the scope of visible structures increases, but the labeling density cannot be sufficiently high to reveal fine structures (ultrastructure)
Solution Approach 1:
By expanding the sample physically, the patent creates additional spatial room that allows high-density labeling to be achieved. The expansion distributes molecules over a larger volume, reducing steric hindrance and enabling higher labeling densities that can reveal ultrastructural details.
Solution Approach 2:
The patent performs physical expansion before labeling, which prepares the sample by creating sufficient spatial separation between molecules. This preliminary action enables subsequent high-density labeling to successfully reveal fine structures that would be impossible to resolve at native density.
3Manufacturing precision
If electron microscopy is used to obtain three-dimensional images of sample ultrastructure, then fine structures can be resolved, but days to weeks of continuous data acquisition are required
Solution Approach 1:
The patent replaces the electron microscopy mechanical imaging system with a light microscopy system combined with physical expansion. This substitution eliminates the need for lengthy electron microscopy data acquisition while achieving comparable ultrastructural resolution through optical means on expanded samples.
Solution Approach 2:
By changing the size parameter of the sample through physical expansion, the patent transforms sub-resolution ultrastructures into super-resolution features visible by light microscopy. This parameter change reduces acquisition time from days/weeks to minutes/hours while maintaining ultrastructural detail.
4Manufacturing precision
If physical expansion of the sample is performed, then fine structures are enlarged to a resolvable scale, but the sample requires embedding in a polymer network and additional processing steps
Solution Approach 1:
The patent introduces a polymer network as an intermediary medium that embeds the biological sample. This intermediary provides mechanical support during expansion, maintains structural integrity, and enables the physical expansion process while preserving the sample's ultrastructural details for subsequent 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
Enables visualization of cellular ultrastructures at high resolution and contrast, overcoming the limitations of fluorescence microscopy and reducing the time required for imaging.
Implementation Method 1
physically expanding the sample by at least a factor of two in at least one dimension
Implementation Method 2
bulk labeling a plurality of components of the sample with at least one reagent to introduce contrast
Data Source
AI summary
Methods and systems for physical expansion and imaging of biological samples are described herein. In one aspect of the disclosure, a method for preparing a biological sample for the purpose of generating images of its ultrastructure with an imaging instrument includes a) physically expanding the sample by at least a factor of two in at least one dimension; and b) bulk labeling a plurality of components of the sample with at least one reagent to introduce contrast.