Detection Disc Focus Groove for Microscopic Object Analysis
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Solution Overview
Problem
Current methods like flow cytometry struggle to detect and count small microscopic objects, such as single molecules, due to background signal interference and prolonged analysis times when dealing with large sample volumes, especially in samples like rolling circle products (RCPs) produced in biotechnology.
Innovation Solution
A detection disc with a focus structure groove and channel, manufactured using a silicon substrate with a crystallographic structure, where the focus structure has sloping sidewalls and a V-shaped cross-section, is used to enhance focusing precision and reduce background noise, allowing for accurate detection and counting of small objects in a fluid sample.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If flow cytometry is used to detect microscopic objects, then integrated fluorescence measurement is achieved, but background signal from free labels in solution overwhelms the signal from objects of interest
Solution Approach 1:
The invention transitions from measuring integrated fluorescence in a three-dimensional volume to detecting fluorescence at a two-dimensional focal plane. The focus structure (groove) creates a specific optical path that allows the detector to observe only objects at a defined depth, effectively filtering out background signal from free labels in solution that are out of focus.
Solution Approach 2:
The detection volume is segmented into a specific focal plane using the groove structure. This divides the three-dimensional sample volume into discrete two-dimensional detection planes, allowing selective observation of objects at specific depths while excluding background signals from other regions.
2Measurement precision
If large sample volumes are analyzed to detect single molecules, then detection sensitivity is improved, but analysis time increases significantly
Solution Approach 1:
By confining detection to a two-dimensional focal plane rather than analyzing the entire three-dimensional sample volume, the invention enables rapid scanning and detection. The groove structure creates a thin optical section that can be quickly imaged, reducing the time required to analyze samples while maintaining sensitivity for single molecule detection.
3Measurement precision
If the detection volume is reduced to improve signal-to-background ratio, then single object detection capability is enhanced, but the ability to detect objects in solution is compromised
Solution Approach 1:
The invention creates a thin two-dimensional detection plane using the groove structure, which allows selective observation of objects at a specific depth. This enables single object detection with high signal-to-background ratio while the entire sample volume can still be analyzed by moving the detection plane or processing multiple grooves, thus maintaining coverage of the total sample quantity.
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 solution enables precise focusing and efficient detection of single small objects, including single molecules, by using a dedicated focusing camera to determine the focus depth for a line detector, significantly reducing analysis time and improving signal-to-background ratio.
Implementation Method 1
the focus structure has sloping sidewalls and a V-shaped cross-section, is used to enhance focusing precision
Implementation Method 2
Flow cytometry is a laser-based, biophysical technology employed in cell counting, cell sorting, biomarker detection
Data Source
AI summary
A substrate for use in manufacture of a production master plate for production of a detection disc for carrying samples in an apparatus for detection of microscopic objects in a fluid, the substrate having a channel and separate focus structure, wherein the focus structure is a groove.