Iterative Expansion Microscopy for High-Throughput Super-Resolution Imaging
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current super-resolution imaging techniques are slow and lack high-throughput capabilities for imaging protein architectures over whole organs with nanoscale resolution, limiting the understanding of diseases such as brain disorders and cancers.
Innovation Solution
Iterative expansion microscopy (iExM) method, which involves multiple iterations of expanding biological samples using swellable materials with different crosslinkers to achieve up to 17- to 19-fold linear expansion, enabling high-throughput super-resolution imaging with nanoscale resolution using high-throughput microscopes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional super-resolution imaging techniques are used, then nanoscale resolution can be achieved, but imaging speed is slow and throughput is limited
Solution Approach 1:
The patent applies expansion microscopy to physically expand the sample in the spatial dimension, transforming a 2D/3D structure into a larger-scale structure that can be imaged at nanoscale resolution using conventional high-throughput microscopes. This dimensional transformation enables simultaneous achievement of both high resolution and high throughput by moving the problem from the optical resolution limit to a physical scale problem.
Solution Approach 2:
The patent introduces an expandant material as an intermediary substance that infiltrates the sample and causes controlled expansion. This intermediary enables the sample to be enlarged without altering its molecular structure, thereby allowing high-throughput imaging at nanoscale resolution that would otherwise be unachievable with conventional techniques.
2Measurement precision
If expansion microscopy is used to improve resolution, then nanoscale detail is enhanced, but the imaging process becomes more complex
Solution Approach 1:
The patent segments the expansion microscopy process into distinct modular steps: sample preparation, expandant infiltration, expansion, and imaging. This segmentation allows each step to be optimized independently and facilitates high-throughput processing by standardizing the workflow, thereby reducing overall process complexity despite the added expansion step.
Solution Approach 2:
The patent utilizes controlled parameter changes in the expandant material properties (such as expansion ratio, gelation conditions, and crosslinking density) to achieve consistent and reproducible sample expansion. By optimizing these parameters, the method achieves high resolution while maintaining process simplicity and reliability through standardized protocols.
3Measurement precision
If iterative expansion is applied to achieve higher expansion ratios, then super-resolution capability is improved, but the number of process steps increases
Solution Approach 1:
The patent implements iterative expansion by nesting multiple expansion cycles within a single workflow, where each expansion step builds upon the previous one. The sample undergoes sequential expansions (e.g., 2-fold followed by another 2-fold expansion) to achieve cumulative high expansion ratios (4-fold or higher). This nested approach enables super-resolution capability while managing process complexity through systematic repetition of standardized steps.
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
iExM provides improved high-throughput super-resolution imaging of protein architectures over whole organs with nanoscale resolution, enhancing the ability to dissect disease mechanisms by allowing for detailed visualization of biological structures beyond the classical diffraction limit.
Implementation Method 1
biological samples of interest are permeated with a swellable material that results in the sample becoming embedded in the swellable material, and then the sample can be expanded isotropically in three dimensions
Data Source
AI summary
The present invention leverages the techniques for expansion microscopy (ExM) to provide improved high-throughput super-resolution whole-organ imaging methodology to image protein architectures over whole organs with nanoscale resolution by using high-throughput microscopes in combination with samples that have been iteratively expanded more than once, in a method referred to herein as “iterative expansion microscopy” (iExM). In the ExM method, biological samples of interest are permeated with a swellable material that results in the sample becoming embedded in the swellable material, and then the sample can be expanded isotropically in three dimensions The process of iteratively expanding the samples can be applied to samples that have been already expanded using ExM techniques one or more additional times to iteratively expand them such that, for example, a 5-fold expanded specimen can be expanded again 3- to 4-fold, resulting in as much as a 17- to 19-fold or more linear expansion.