Single Cell Analysis Device with Segmented Porous Membrane
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Solution Overview
Problem
The existing cell analysis devices face challenges in efficiently capturing nucleic acids while maintaining high cell capturing efficiency, as the improvement in nucleic acid capturing efficiency leads to increased pressure loss, resulting in reduced suction rates and cell settlement, and the amplification products often adsorb on the device walls, affecting the accuracy of single cell analysis.
Innovation Solution
The device employs a repulsive force to prevent cells from settling on the substrate other than the cell capture part, using gravity or electrostatic forces, and incorporates a three-dimensional porous membrane with a hydrophilic inner wall to reduce adsorption, along with a separation mechanism to prevent amplification products from reaching the inner wall, such as using air or nonpolar solvents and ultrafiltration membranes.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If the average pore diameter of the porous membrane is reduced to several μm or less (especially 1 μm or less) and the thickness is increased to 10 μm or more (especially several tens of μm or more) to improve nucleic acid capturing efficiency, then the nucleic acid capturing efficiency is improved, but the pressure loss increases and the suction rate decreases
Solution Approach 1:
The device is divided into two distinct functional regions: a cell capture part with larger pores for efficient cell suction and a nucleic acid capture part with smaller pores for efficient nucleic acid capture. This segmentation allows each region to be optimized for its specific function without compromise
Solution Approach 2:
Different pore sizes are applied to different locations of the porous membrane: larger pore diameters in the cell capture part region and smaller pore diameters in the nucleic acid capture part region. This local quality variation resolves the contradiction by providing appropriate pore characteristics for each functional requirement
2Measurement precision
If the porous membrane thickness is increased to 10 μm or more (especially several tens of μm or more) to improve nucleic acid capturing efficiency, then the nucleic acid capturing efficiency is improved, but the pressure loss increases and the suction rate decreases
Solution Approach 1:
The device is divided into two distinct functional regions: a cell capture part with larger pores for efficient cell suction and a nucleic acid capture part with smaller pores for efficient nucleic acid capture. This segmentation allows each region to be optimized for its specific function without compromise
Solution Approach 2:
Different pore sizes are applied to different locations of the porous membrane: larger pore diameters in the cell capture part region and smaller pore diameters in the nucleic acid capture part region. This local quality variation resolves the contradiction by providing appropriate pore characteristics for each functional requirement
3Measurement precision
If the pressure loss is reduced by using smaller pore diameter and greater thickness, then the nucleic acid capturing efficiency is improved, but the suction rate of cells into the cell capture part decreases
Solution Approach 1:
The device is divided into two distinct functional regions: a cell capture part with larger pores for efficient cell suction and a nucleic acid capture part with smaller pores for efficient nucleic acid capture. This segmentation allows each region to be optimized for its specific function without compromise
Solution Approach 2:
Different pore sizes are applied to different locations of the porous membrane: larger pore diameters in the cell capture part region and smaller pore diameters in the nucleic acid capture part region. This local quality variation resolves the contradiction by providing appropriate pore characteristics for each functional requirement
4Reliability
If cells remain in regions other than the cell capture part due to gravity settlement, then the ratio of analyzable cells decreases and nucleic acid contamination increases, but preventing this requires reducing pressure loss which conflicts with nucleic acid capturing efficiency
Solution Approach 1:
The device is divided into two distinct functional regions: a cell capture part with larger pores for efficient cell suction and a nucleic acid capture part with smaller pores for efficient nucleic acid capture. This segmentation allows each region to be optimized for its specific function without compromise
Solution Approach 2:
Different pore sizes are applied to different locations of the porous membrane: larger pore diameters in the cell capture part region and smaller pore diameters in the nucleic acid capture part region. This local quality variation resolves the contradiction by providing appropriate pore characteristics for each functional requirement
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
This approach enhances cell capturing efficiency and maintains nucleic acid capturing efficiency, allowing for accurate single cell analysis by preventing cell adsorption and ensuring high-yield amplification products, enabling precise gene expression analysis at the single cell level.
Implementation Method 1
the substrate has a repulsive force in a direction which separates the cells from the substrate
Implementation Method 2
the substrate has a repulsive force in a direction which separates the cells from the substrate
Implementation Method 3
a three-dimensional porous membrane with a hydrophilic inner wall to reduce adsorption
Implementation Method 4
a three-dimensional porous membrane with a hydrophilic inner wall to reduce adsorption
Implementation Method 5
using air or nonpolar solvents and ultrafiltration membranes
Implementation Method 6
the pressure control means controls so that when the cell is captured by the cell capture part, a force in a direction from the cell capture part to the nucleic acid capture part acts as a first pressure
Data Source
AI summary
The purpose of the present invention is to provide a single cell analysis device in which the improvement of the nucleic acid capturing efficiency and the improvement of the cell capturing efficiency are both achieved and a highly accurate single cell analysis data is thereby obtained. The present invention relates to an improvement of a cell analysis device including a two-dimensional array chip having a plurality of cell capture parts capable of capturing a single cell in each of the capture parts, and nucleic acid capture parts corresponding to the respective cell capture parts, the nucleic acid capture parts being capable of capturing a nucleic acid extracted from the cell captured by the cell capture part.