DNA-Protein-Polymer-Metal Cation Complex for High-Resolution SEM Imaging
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Solution Overview
Problem
Current microscopy techniques, such as transmission electron microscopy (TEM) and fluorescence microscopy (FM), face limitations in imaging DNA molecules with high resolution and compatibility with microfluidic devices.
Innovation Solution
A DNA-protein-polymer-metal cation complex is developed, which enables high-resolution, real-time imaging of DNA molecules under scanning electron microscopy (SEM) by enhancing the resolution and compatibility with microfluidic devices.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If TEM is used to observe DNA molecules, then detailed observation of DNA fine structures is enabled, but the procedure becomes time-consuming and complex due to the need for heavy metal salts and shadow casting techniques
Solution Approach 1:
The patent extracts and eliminates the complex shadow casting procedure and heavy metal salt staining steps from the TEM imaging process. By using DNA-binding proteins with intrinsic electron scattering capability, the method removes the need for additional staining and shadow casting steps, significantly simplifying the procedure while maintaining high-resolution observation of DNA fine structures
Solution Approach 2:
The patent introduces DNA-binding proteins as intermediaries that directly bind to DNA molecules and provide electron scattering contrast. These proteins serve as a bridge between DNA and the electron beam, enabling direct visualization without requiring heavy metal salts or shadow casting techniques, thus resolving the contradiction between observation detail and procedural complexity
2Ease of operation
If fluorescence microscopy is used for DNA imaging, then simplicity and accessibility are achieved, but resolution is insufficient for accurate observation at the nanometer scale
Solution Approach 1:
The patent changes the detection parameter from optical fluorescence to electron scattering. By using DNA-binding proteins that scatter electrons, the method achieves nanometer-scale resolution comparable to TEM while maintaining the simplicity of fluorescence microscopy workflows, including compatibility with microfluidic devices and chemically functionalized surfaces
Solution Approach 2:
The patent creates a multi-functional imaging approach that combines the ease of fluorescence microscopy operations with the resolution of electron microscopy. The DNA-binding proteins enable the system to function both as a simple fluorescent assay platform and as a high-resolution electron microscopy sample, eliminating the need to choose between simplicity and precision
3Measurement precision
If TEM is used for DNA imaging, then high resolution is achieved, but compatibility with microfluidic devices and chemically functionalized surfaces is poor due to alignment issues with carbon film-coated metal grids
Solution Approach 1:
The patent inverts the traditional TEM approach by instead of placing DNA on carbon film-coated metal grids, it uses DNA-binding proteins that can be directly applied to microfluidic devices and chemically functionalized surfaces. This inversion allows DNA imaging to be performed in microfluidic channels and on functionalized surfaces, achieving both high resolution and compatibility with modern molecular biology platforms
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
The complex allows for enhanced visualization of DNA molecules with diverse shapes and lengths at high magnification, improving DNA optical mapping and facilitating real-time imaging.
Implementation Method 1
a polymer, an anhydride thereof, or a salt thereof, which is capable of binding to the DNA-binding protein via intermolecular electrostatic interaction
Implementation Method 2
a metal cation-containing component, excluding uranium, that is capable of binding to the polymer, the anhydride thereof, or the salt thereof through interaction
Implementation Method 3
a DNA-binding protein comprising a peptide having an amino acid sequence capable of binding to the DNA molecule
Implementation Method 4
having an amino acid sequence capable of binding to the DNA molecule
Data Source
AI summary
Disclosed are a DNA-protein-polymer-metal cation complex, a DNA imaging method using the same, and a scanning electron microscopy (SEM)-based DNA detection method. The DNA-protein-polymer-metal cation complex disclosed herein enables real-time imaging of DNA molecules of various shapes and lengths with improved resolution by using SEM even at high magnifications, and improves DNA detection, thereby further enhancing DNA optical mapping.