Micro-chamber plate for real-time PCR with transparent sealing
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Solution Overview
Problem
Current micro-chamber plates for real-time PCR face challenges such as high sample volume requirements, cross-contamination, and bubble formation during temperature cycling, which complicate the analysis of multiple samples and lead to measurement errors.
Innovation Solution
A micro-chamber plate design featuring a transparent layer for optical measurement, an injection layer with punched holes for efficient reagent delivery, and a sample supply layer to prevent evaporation and bubble formation, allowing for easy sample injection and simultaneous analysis of a large number of samples without cross-contamination.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If a standard 96-well plate or 384-well plate is used for real-time PCR analysis, then multiple samples can be analyzed simultaneously, but the sample volume required is large (10-50 μl per well)
Solution Approach 1:
The plate is divided into multiple independent micro-chambers (1536 chambers per plate) instead of using traditional large wells. Each micro-chamber contains only the necessary reaction components (primers, probes, buffer) and requires minimal sample volume (1-5 μl), enabling high-throughput analysis with reduced sample consumption.
Solution Approach 2:
The invention transitions from a 2D array of large wells to a 3D micro-chamber structure with vertical depth, allowing dense packing of 1536 micro-chambers in a single plate while maintaining optical measurement capability through transparent materials and optimized chamber geometry.
2Reliability
If micro-chambers are sealed with a semitransparent membrane to isolate reaction solutions, then cross-contamination is prevented, but evaporation of reaction solution occurs and additional sealing steps are required
Solution Approach 1:
A flexible transparent film is used to seal the micro-chambers, providing both isolation to prevent cross-contamination and optical transparency for fluorescence measurement. The film creates a closed system that prevents evaporation while maintaining measurement capability.
Solution Approach 2:
The plate structure combines transparent plastic or glass materials that provide both optical clarity for fluorescence detection and barrier properties to prevent evaporation and cross-contamination, integrating multiple functions into a single material system.
3Loss of substance
If a water-repellent membrane is inserted between cover glass and wafer to prevent evaporation, then evaporation is reduced, but the process becomes troublesome requiring separation and elimination of multiple layers
Solution Approach 1:
The invention removes the complex multi-layer structure (cover glass, water-repellent membrane, wafer) and extracts only the essential sealing and measurement functions, implementing them in a single integrated transparent plate with micro-chambers that requires no layer separation or elimination steps.
Solution Approach 2:
The transparent plate structure is designed to be self-sealing through its integrated micro-chamber geometry and flexible film, eliminating the need for additional water-repellent membranes or complex assembly/disassembly procedures. The structure serves its own sealing and measurement functions without external assistance.
4Reliability
If mineral oil is used to seal micro-chambers completely, then cross-contamination is prevented, but bubbles are formed during temperature cycling causing measurement errors
Solution Approach 1:
A flexible transparent film seals the micro-chambers without introducing bubbles, providing both cross-contamination prevention and optical transparency. The film conformally seals the chamber geometry without trapping air, eliminating the bubble formation problem associated with mineral oil while maintaining measurement accuracy.
5Manufacturing precision
If different solutions are loaded in each micro-chamber using a micropipette, then precise delivery is achieved, but the process takes a long time especially for 1,536 micro-chambers
Solution Approach 1:
The invention combines multiple solution delivery functions into a single integrated micro-chamber plate design where all micro-chambers are manufactured with precise geometry and sealing characteristics, eliminating the need for individual micropipetting of each chamber. The plate can be loaded as a single unit or in batches, dramatically increasing throughput while maintaining precision through standardized chamber design.
Data Source
AI summary
The present invention relates to a micro-chamber plate and a manufacturing method of the same, more precisely a micro-chamber plate facilitating real-time measurement and analysis of fluorescence obtained from the reaction of multiple reaction solutions containing primers or probes selectively binding to each corresponding gene without cross-contamination in order to analyze biological samples containing numbers of genes and a manufacturing method of the same.