Tunable Filter Optical Path for Bead Assay Localization
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Solution Overview
Problem
Existing microscopy systems for bead-based assays are computationally intensive and prone to errors due to the need for multiple sensors, which increases processing time and susceptibility to optical alignment issues when localizing and analyzing luminescently labeled microspheres at multiple emission wavelengths.
Innovation Solution
An optical imaging system with a tunable filter is used to localize microspheres based on back-scattered light and read emission spectra, eliminating the need for additional sensors by sharing a common optical path for reflected light and luminescent emission, and allowing for the identification and quantification of target molecules attached to the beads.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If multiple sensors are used to measure forward scatter and luminescence signals, then measurement capability is improved, but device complexity and susceptibility to optical alignment problems increase
Solution Approach 1:
The patent combines forward scatter measurement and luminescence detection into a single sensor system. The detector captures both the forward scatter signal (for microsphere localization) and luminescence signals (for spectral identification) through a common optical path, eliminating the need for separate sensors and reducing optical alignment complexity
Solution Approach 2:
The single detector is designed to perform multiple functions: it measures forward scatter intensity for positioning microspheres, detects luminescence at multiple wavelengths for spectral coding identification, and can operate across different emission wavelengths. This multi-functional approach replaces multiple specialized sensors with one versatile detection system
2Measurement precision
If multispectral images are acquired at multiple emission wavelengths, then spectral identification accuracy is improved, but processing time increases proportionally with the number of colors
Solution Approach 1:
The system performs preliminary localization of all microspheres in the field of view using forward scatter measurements before acquiring luminescence images. This preliminary positioning step creates a reference map that enables efficient subsequent analysis, allowing the system to process multiple wavelengths without proportionally increasing overall processing time
Solution Approach 2:
The patent implements continuous scanning across multiple emission wavelengths without requiring complete re-localization at each wavelength. The forward scatter-based localization serves all wavelength acquisitions, and the system continuously accumulates spectral data from the pre-localized positions, maintaining efficient workflow throughout the multispectral imaging process
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 reduces the number of images required for analysis, decreases processing time, and minimizes calibration errors, enabling faster and more robust bead-based assays with improved accuracy and efficiency.
Implementation Method 1
localize microspheres in bead-based assays based on a back-scattered light image
Implementation Method 2
read emission spectra from the microspheres
Implementation Method 3
images of assayed microspheres are acquired at multiple emission wavelengths. The luminescent spectral code and fluorescence from each microsphere
Data Source
AI summary
This invention relates to a system and method related to an epifluorescence microscope based optical system equipped with a tunable filter to localize microspheres in bead-based assays based on a back-scattered light (also known as reflected light) image. A common optical path for reflected and emitted luminescence in conjunction with a tunable filter negates the requirement of an additional sensor employed in existing technologies for localizing microspheres based on light scatter measurements.