Ligand Binding Assays on Multiwell Plate Sidewalls
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Solution Overview
Problem
Current molecular binding event characterization systems, such as those using TIR imaging ellipsometry, are limited in scalability and prone to sedimentation issues when imaging larger areas like 384 well and 96 well plates, requiring complex optics and being sensitive to sediment falling onto the detection area.
Innovation Solution
The system immobilizes ligand arrays on the side walls of multiwell plates, allowing polarized light to generate a TIR evanescent field without prisms or gratings, enabling larger area imaging and reducing sedimentation by orienting ligands upright, thus simplifying the setup and increasing cost-effectiveness.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Area of stationary object
If TIR imaging ellipsometry is used to image larger areas like 384 well and 96 well plates, then the measurement area is increased, but the device complexity increases due to requiring prisms or gratings and the sedimentation problem worsens
Solution Approach 1:
The patent extracts and eliminates the prism or grating components from the optical system. By directing polarized light through the transparent plate material between the wells to achieve TIR at the sidewalls, the system removes the need for complex coupling optics while maintaining the evanescent field generation capability across large areas.
Solution Approach 2:
The patent transitions from imaging ligand arrays on horizontal surfaces to imaging arrays on vertical sidewalls. This dimensional change allows the light to pass through the plate between wells and strike the sidewalls at the appropriate angle for TIR, eliminating the need for prisms while enabling large-area measurement.
2Ease of operation
If ligand arrays are placed at the bottom of open wells for imaging, then the detection area is accessible, but sedimentation onto the detection area increases
Solution Approach 1:
The patent moves the ligand array from the horizontal bottom surface to the vertical sidewall of the well. This repositioning changes the orientation of the detection area from horizontal to vertical, allowing gravity to prevent sediment from falling onto the ligand array while maintaining accessibility for imaging through the transparent plate.
3Reliability
If prisms or gratings are used to achieve TIR for large area imaging, then the evanescent field is generated, but the cost and device complexity increase
Solution Approach 1:
The patent removes the prism or grating components from the system. By utilizing the transparent plate material itself as the medium through which polarized light passes to achieve TIR at the sidewalls, the system generates the evanescent field without requiring additional optical coupling components, thereby reducing complexity and cost.
Solution Approach 2:
The transparent plate material serves multiple functions: it contains the wells, provides structural support, and acts as the optical medium through which polarized light passes to generate TIR at the sidewalls. This multi-functionality eliminates the need for separate prism or grating components.
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
This approach allows for cost-effective scaling of molecular binding event characterization across larger areas, reducing sedimentation issues and eliminating the need for complex optics, while maintaining high sensitivity and accuracy in ligand binding assays.
Implementation Method 1
a beam of polarized light is directed through the transparent plate material between the wells in a manner to achieve TIR and an evanescent field in the plane of the ligands
Data Source
AI summary
An apparatus and method for real time, label-free imaging and quantitation of binding events at an array of positions are provided. Total internal reflection from a planar side wall of a well of a multiwell plate is used to create an evanescent field in the plane of a pattern of ligands immobilized on the wall. Embodiments include imaging and multiple analyte detection and quantitation of a single wall of a single well as well as the simultaneous imaging and multiple analyte detection and quantitation of a number of wells.