Microwell Plate Partitioning for Precise Digital Assays
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Solution Overview
Problem
Existing biological and chemical assays face challenges in achieving accurate, simple, and fast analyte detection, particularly for low concentrations, due to inefficiencies in sample partitioning and signal amplification.
Innovation Solution
A device comprising movable plates with microwells of predetermined geometry and volume, allowing for precise sample partitioning and signal amplification through closed configurations, which enhance analyte detection by minimizing cross-well interference and enabling digital PCR techniques.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If samples are partitioned into microwells with predetermined geometry and volume, then measurement precision and quantification accuracy are improved, but device complexity increases due to the need for precise microwell fabrication and plate alignment mechanisms
Solution Approach 1:
The sample is partitioned into multiple isolated microwells with predetermined geometry and volume on the second plate. Each microwell acts as an independent reaction chamber, enabling digital PCR assays where individual analyte molecules can be detected and counted. This segmentation provides absolute quantification of nucleic acids without requiring standard curves, directly improving measurement precision.
Solution Approach 2:
The device utilizes controlled spacing parameters between the first plate inner surface and microwell rims, transitioning from an open configuration (spacing > well depth) for sample deposition to a closed configuration (spacing < 1 μm) for assay execution. This parameter change enables precise volume control and isolation of samples in micrawells while maintaining ease of operation.
2Ease of operation
If the device uses movable plates with open and closed configurations, then ease of operation is improved for sample deposition, but reliability may worsen due to potential misalignment or contamination between configurations
Solution Approach 1:
The device employs movable first and second plates that can transition between open and closed configurations. In the open configuration, plates are spaced apart to allow droplet deposition onto the micrawell plate. In the closed configuration, plates are brought together with spacing less than 1 μm to seal the samples in micrawells. This dynamic design enables easy sample loading while maintaining reliable sample isolation during assays.
Solution Approach 2:
The spacing between the first plate inner surface and micrawell rims acts as an intermediary mechanism. In the open configuration, larger spacing allows free access for sample deposition. In the closed configuration, the spacing reduces to less than 1 μm (or less than 1/10 of micrawell depth), effectively sealing the samples without requiring direct contact between plates, thus preventing contamination while maintaining isolation.
3Productivity
If micrawells have small volume substantially less than the fluidic sample, then productivity is improved by enabling parallel processing of multiple samples, but manufacturing precision requirements increase for consistent micrawell fabrication
Solution Approach 1:
The fluidic sample is divided into multiple micrawells, each with a volume substantially less than the total sample volume. This segmentation enables parallel processing of multiple partitions simultaneously in a single device, significantly improving productivity. Each micrawell can independently contain and process individual analyte molecules or droplets, allowing high-throughput digital PCR assays.
Solution Approach 2:
The micrawells are designed with predetermined and known geometry parameters, including specific depth (200 μm or less) and volume constraints. These controlled geometric parameters ensure consistent partitioning of samples across all micrawells, enabling reliable quantitative measurements while maintaining manufacturability through standardized fabrication processes.
Data Source
AI summary
Provided herein is a method and device for partitioning a fluidic sample. The device contains a plate containing microwells. The method comprises depositing a sample on one or both of the plates when the plates are in an open configuration, wherein the deposition is in the form of a single or multiple droplet of the sample, wherein at least one of the droplets has a volume that occupies more than two microwells and closing the plates to the closed configuration to partition the sample in the microwells.