Negative-Pressure Dot Immunoblotting for Multi-Sample Detection
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Solution Overview
Problem
Existing protein detection methods like Western blot and ELISA are complex, require large sample quantities, and lack sensitivity, specificity, and the ability to perform multi-sample detection simultaneously.
Innovation Solution
A dot immunoblotting detection device and method using a negative pressure suction device, a hole plate with 70 through holes, and a nitrocellulose membrane, allowing for sample concentration and formation of injection grooves, followed by a series of immunoreactions and image analysis to determine protein expression levels.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If Western blot or ELISA methods are used for protein detection, then detection sensitivity and specificity can be achieved, but the operation becomes complicated and requires a large number of samples
Solution Approach 1:
The device segments the detection process by using a plate with multiple through-holes, allowing simultaneous processing of multiple samples in parallel. Each hole acts as an independent detection unit, dividing the complex single-sample Western blot/ELISA process into multiple simple parallel operations, thereby reducing overall operation complexity while maintaining detection sensitivity and specificity.
Solution Approach 2:
The detection device is designed as a universal platform that can handle multiple sample types and detection conditions simultaneously. The plate structure with standardized through-holes allows it to perform both Western blot and ELISA-type detections, making it a multi-functional device that reduces the need for separate complex procedures for different detection needs.
2Measurement precision
If Western blot or ELISA methods are used for protein detection, then detection accuracy is maintained, but the ability to perform multi-sample detection simultaneously is lost
Solution Approach 1:
The plate is divided into multiple through-holes, each capable of holding and processing a separate sample independently. This segmentation allows simultaneous multi-sample detection while maintaining the detection accuracy of individual samples, as each hole provides an isolated environment similar to traditional single-sample methods.
Solution Approach 2:
The invention transitions from single-sample sequential detection to multi-sample parallel detection by adding the spatial dimension of multiple holes in the plate. This dimensional change allows multiple samples to be processed simultaneously without compromising the vertical depth of the immunodetection reaction, thereby maintaining accuracy while increasing productivity.
3Ease of operation
If traditional detection methods are used, then sample processing is straightforward, but cross-contamination occurs and reduces detection reliability
Solution Approach 1:
Each through-hole in the plate provides a physically separated compartment for sample processing. This segmentation prevents cross-contamination between samples while maintaining ease of operation, as samples remain isolated in their respective holes throughout the detection process, yet the overall procedure remains simple and straightforward.
Solution Approach 2:
The plate structure acts as an intermediary barrier between samples, preventing direct contact and cross-contamination. This intermediary structure allows simple sample processing procedures to be performed without compromising reliability, as the physical separation provided by the plate holes blocks contamination pathways while maintaining operational simplicity.
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 method provides sensitive, specific, and economical multi-sample detection with reduced cross-contamination, suitable for large-scale testing.
Implementation Method 1
a negative pressure suction device, a hole plate with 70 through holes, and a nitrocellulose membrane, allowing for sample concentration and formation of injection grooves
Data Source
AI summary
The present invention provides a detection device of dot immunoblotting detection, including a negative pressure suction device, a hole plate, a hose for connecting the hole plate with the negative pressure suction device, and a nitrocellulose membrane in tight fit with the upper end surface of the hole plate. The present invention further provides a detection method of dot immunoblotting detection, including preparation, sample injection, blocking, incubation of a primary antibody, incubation of a secondary antibody, development and analysis. The detection method performs negative pressure suction through the negative pressure suction device, which is favorable for concentrating samples during sample injection and avoiding the influence, caused by cross contamination after diffusion of the samples, on an experimental result, and the experimental result is more accurate.