Flat-Field Imaging Alignment for In-Focus Target Separation
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Solution Overview
Problem
Existing methods for aligning and imaging colloidal and cellular suspensions or droplets are cumbersome and lack real-time feedback, making efficient extraction and analysis difficult.
Innovation Solution
A method involving the combination of three immiscible phases, where a third phase containing targets is positioned between two other phases, allowing for alignment and imaging in a flat-field configuration using an optical sensor.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If traditional FACS systems are used for cell sorting and manipulation, then cells can be separated and manipulated in continuous microfluidic flows, but the system becomes complex and cumbersome
Solution Approach 1:
The system divides the sample into discrete droplets containing individual cells or particles, with each droplet acting as an independent unit. This segmentation allows parallel processing of multiple samples simultaneously, reducing overall system complexity while maintaining high throughput capability
Solution Approach 2:
The patent introduces an intermediary imaging system that captures images of droplets in a flat-field configuration, serving as a mediator between the microfluidic flow and the sorting mechanism. This intermediary step simplifies the control system by providing direct visual feedback for real-time sorting decisions
2Measurement precision
If conventional imaging methods are used for target alignment, then imaging can be performed, but targets exhibit significant overlap and are not in focus within the field of view
Solution Approach 1:
The patent transitions from conventional wide-field imaging to a confocal or optical sectioning approach that isolates a specific focal plane. By imaging only at the focal plane where droplets are in focus, the system eliminates out-of-focus overlap and achieves precise measurement of individual targets
Solution Approach 2:
The system ensures that only targets within the focal plane are imaged with high quality, while targets outside this plane are excluded from the image. This local quality approach maintains sharp focus and prevents overlap by selectively capturing only in-focus targets
3Productivity
If conventional alignment methods are used, then particle positioning can be attempted, but real-time feedback is lacking and analysis efficiency is reduced
Solution Approach 1:
The patent implements a closed-loop feedback system where the imaging system continuously monitors droplet positions and characteristics in real-time, and this information is immediately used to control the sorting mechanism. This real-time feedback eliminates delays and enables rapid sorting decisions
Solution Approach 2:
The system maintains continuous imaging and sorting operations without interruption, with the imaging system continuously capturing droplet information as they flow through the field of view. This continuous operation eliminates idle time and maintains maximum productivity throughout the sorting 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
Enables rapid and accurate imaging of targets with minimal overlap, improving analysis and sorting efficiency by ensuring targets are in focus within the field of view.
Implementation Method 1
combining a first phase, a second phase, and the third phase in a volume... the first phase, the second phase, and the third phase may be substantially immiscible, and the third phase may be in fluid communication with the first phase and the second phase
Data Source
AI summary
A method of aligning a plurality of targets is provided. The method includes generating a plurality of targets. A third phase includes the plurality of targets. The method further includes combining a first phase, a second phase, and the third phase in a volume. The first phase, the second phase, and the third phase are substantially immiscible, and the third phase is in fluid communication with the first phase and the second phase, and the first phase, the second phase, and the third phase are operable to be in a configuration of the third phase between the first phase and the second phase in the volume.