Porous Insert Structure for Convection-Suppressed Spatial Profiling
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current spatial profiling methods for target analytes in biological samples suffer from convection-induced mislocalization during processing, leading to discrepancies between the native position of analytes in tissues and their captured positions on detection arrays, which compromises spatial resolution and accuracy.
Innovation Solution
The use of a porous insert structure with a hydrophilic surface, placed in the assay chamber to limit free flow space and reduce fluid convection, combined with a capture probe having a spatial barcode to correlate and identify the location of biological analytes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If a tissue is permeabilized by proteinase in a solution and mRNA transcripts are released to migrate to a proximal location on a spatial array by diffusion and gravity, then the analyte can be captured on the detection slide, but convection during processing steps causes mislocalization of the analyte away from its native position
Solution Approach 1:
A porous insert structure is introduced into the assay chamber to limit free flow space and reduce fluid convection. The porous material allows controlled permeation while restricting bulk fluid movement that would cause convection-induced analyte displacement, thereby improving spatial resolution and localization accuracy.
Solution Approach 2:
The porous insert acts as an intermediary element between the tissue sample and the detection array. It mediates the fluid dynamics by limiting convection currents while still allowing necessary permeabilization and analyte migration, thus resolving the conflict between processing requirements and spatial accuracy.
2Ease of operation
If the free flow space in the chamber is increased to allow adequate permeabilization solution penetration, then the tissue can be properly processed, but convection currents increase and cause greater analyte displacement
Solution Approach 1:
The porous insert provides a structure that maintains adequate permeabilization solution penetration through its porous network while simultaneously limiting bulk fluid flow and convection currents. This resolves the contradiction by decoupling the requirements for solution penetration from uncontrolled convection.
Solution Approach 2:
The porous insert changes the physical parameters of the chamber by introducing a structure with controlled porosity and surface area. This modifies fluid dynamics to allow necessary permeabilization while restricting convection, thus improving spatial resolution without compromising processing efficiency.
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
Enhances spatial resolution and accuracy in profiling target analytes by minimizing convection-induced mislocalization, allowing for precise correlation of analyte locations within biological samples.
Implementation Method 1
the porous insert limits free flow space in the chamber, thereby reducing fluid convection above the biological sample
Implementation Method 2
An insert structure may be made of porous material, and may include a hydrophilic surface, which allow a permeabilization solution to easily penetrate through the insert structure
Implementation Method 3
capturing a biological analyte from the biological sample or an intermediate agent indicative of the presence of the biological analyte in the biological sample using the capture probe
Data Source
AI summary
Provided herein are technologies for reducing fluid convection during processing and analysis of a biological sample. The provided technologies can include the use of a porous insert to limit fluid convection adjacent to a biological sample. In particular, provided technologies can include providing a substrate comprising a capture area, a biological sample comprising a cell disposed on the capture area, a buffer disposed on the biological sample, and a gasket disposed on the substrate, wherein the gasket provides a chamber comprising the lateral dimension of the capture area and includes a height above the capture area and biological sample; and a porous insert positioned in the chamber and in contact with the buffer and/or or the biological sample, wherein the porous insert limits free flow space in the chamber, thereby reducing fluid convection adjacent to the biological sample.