Microplate with Integrated Microfluidic Channels for TIRF Imaging
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Solution Overview
Problem
Current microplate technologies are inadequate for high-throughput assays that require single-molecule recognition and total internal reflection fluorescence (TIRF) microscopy, as they lack efficient fluidic connectivity and functionalized imaging zones for simultaneous analysis of multiple samples.
Innovation Solution
A microplate design featuring multiple layers with integrated microfluidic channels and a prism for TIRF microscopy, allowing fluid connection between wells and a central imaging zone for simultaneous sample analysis, and functionalized imaging zones for specific analyte binding.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Productivity
If traditional microplate technologies are used, then device simplicity is maintained, but assay throughput and single-molecule recognition capability are insufficient
Solution Approach 1:
The microplate is divided into multiple functional layers: a top layer with sample wells, a middle layer with microfluidic channels for fluid transport, and a bottom layer with imaging zones for detection. This segmentation allows each layer to be optimized for its specific function while working together to achieve high-throughput single-molecule analysis
Solution Approach 2:
The patent implements a nested structure where microfluidic channels are integrated within the microplate layers, and imaging zones are positioned within the bottom layer. Multiple functional elements are nested within the compact microplate architecture, enabling complex functionality without proportionally increasing overall device footprint
2Productivity
If multiple samples are analyzed simultaneously, then productivity increases, but sample volume requirements and reagent consumption increase
Solution Approach 1:
The patent employs microfluidic channels with integrated vacuum ports to control fluid flow. By applying vacuum pressure, samples and reagents are efficiently drawn through the microfluidic network to the imaging zones, enabling precise delivery of small volumes while maintaining high throughput analysis of multiple samples
Solution Approach 2:
The patent transitions from traditional two-dimensional well-based analysis to a three-dimensional microfluidic system with vertical channel networks. This dimensional change allows samples from multiple wells to be routed through channels to shared imaging zones, enabling simultaneous analysis of multiple samples with reduced per-sample reagent volumes
3Productivity
If binding times are reduced, then productivity increases, but measurement precision may be compromised
Solution Approach 1:
The patent replaces traditional mechanical mixing and incubation methods with microfluidic flow control. By using controlled fluid flow through the microfluidic channels, samples are efficiently delivered to imaging zones and binding reactions are initiated with precise temporal control, achieving fast binding times while maintaining detection precision through consistent flow conditions
Solution Approach 2:
The microfluidic system enables continuous flow of samples and reagents through the imaging zones, maintaining continuous binding reactions. This continuous action allows for rapid equilibrium to be reached while ensuring consistent measurement conditions, thereby preserving measurement precision even with reduced binding times
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
Enhances assay throughput, reproducibility, and reduces sample volume requirements by bundling samples into a single imaging zone, enabling faster binding times and cost-effective multiplex analysis with reduced reagent consumption.
Implementation Method 1
single-molecule recognition through equilibrium Poisson sampling (SiMREPS) and in other assays employing total internal reflection fluorescence (TIRF) or HiLo microscopy
Implementation Method 2
Layer 2 lacks the prism and the layer comprises a high refractive index material suitable for use as a waveguide in waveguide TIRF
Data Source
AI summary
This disclosure pertains to a consumable product (microplate) suitable for use in assays utilizing single-molecule recognition through equilibrium Poisson sampling (SiMREPS) and in other assays employing total internal reflection fluorescence (TIRF) or HiLo microscopy. The disclosed microplate is also suitable for use in other high throughput assay systems, such as single-molecule FRET, ligand-receptor binding studies, membrane biology assays, cell-based TIRF and near-TIRF assays. The disclosure further pertains to the use of the microfluidic microplate for the detection of analytes, including nucleic acids, polypeptides, carbohydrates, lipids, post-translational modifications, amino acids, metabolites, and small molecules.