Fluorescence Detection Background Subtraction Resin Microchips
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Solution Overview
Problem
Fluorescence detection methods face accuracy issues due to intense background fluorescence from microchips, which prolong detection times and increase costs, especially with resin microchips where the background fluorescence persists longer than silica glass, necessitating additional treatments like quenchers.
Innovation Solution
A fluorescence detection method that involves detecting and storing the temporal change of fluorescence intensity from a reference microchip, allowing for subtraction of background fluorescence from the biological sample's signal before the background intensity fully attenuates, enabling accurate and rapid detection without additional treatments.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If resin microchips are used to reduce cost, then manufacturing cost is reduced, but background fluorescence intensity increases and detection time increases
Solution Approach 1:
The patent applies preliminary action by measuring the background fluorescence of the resin microchip before sample injection and storing this temporal profile. This pre-measured background data is then subtracted from the actual detection signal, eliminating the need to wait for background attenuation and enabling immediate sample detection while maintaining accuracy.
2Measurement precision
If waiting for background fluorescence to attenuate before detection, then measurement precision is improved, but detection time increases
Solution Approach 1:
The patent extracts the background fluorescence component from the total detected signal by subtracting the pre-measured background temporal profile from the actual detection signal. This separation allows the sample fluorescence to be analyzed independently without waiting for background attenuation, thus improving detection speed while maintaining precision.
Solution Approach 2:
The patent applies preliminary anti-action by pre-measuring and storing the background fluorescence temporal profile before sample detection. This pre-characterized background data is then used to counteract the background interference during actual detection, eliminating the need to wait for attenuation and enabling immediate accurate measurement.
3Measurement precision
If quencher treatment is applied to reduce background, then measurement precision is improved, but device complexity and cost increase
Solution Approach 1:
The patent replaces the chemical approach (quencher treatment) with a computational/mathematical approach. Instead of modifying the microchip physically or chemically to reduce background, the method uses background subtraction algorithms to eliminate background interference from the detected signal, simplifying the overall system while maintaining detection accuracy.
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 method allows for high-accuracy detection of fluorescence intensity from biological samples with reduced background influence and shorter detection times, eliminating the need for costly quencher treatments and reducing the time required for background fluorescence to attenuate.
Implementation Method 1
a fluorescently-labeled biological sample fed to a microchip and excitation light is applied thereto to detect an intensity of emitted fluorescence
Implementation Method 2
the fluorescence emitted from the microchip itself... The fluorescence emitted from the microchip acts as background and lowers the accuracy, i.e., the S/N ratio, of the detection of the fluorescence intensity
Data Source
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Figure 3
Figure 4A~4C
AI summary
A fluorescence detection method is disclosed, which allows detection of a fluorescence intensity from a fluorescently-labeled biological sample with high accuracy in a shorter time, even when a fluorescence intensity from a microchip itself acting as background. A fluorescence intensity (Ur0) of fluorescence (Lk0) emitted from an equivalent microchip (11) itself, which is equivalent to a microchip (10) with a fluorescently-labeled biological sample (R) fed thereto, is detected for a period from the start of application of excitation light (Le) until the fluorescence intensity (Ur0) sufficiently attenuates, and temporal change of the fluorescence intensity (Ur0) is stored. The excitation light (Le) is applied to the microchip (10) and the biological sample (R) is fed before the fluorescence intensity (Ur0) from the microchip (10) itself sufficiently attenuates to detect the emitted fluorescence intensity (Ur) for a period from the start of application of the excitation light (Le) to a point of time after the biological sample (R) is fed. Values of the stored temporal change of the fluorescence intensity (Ur0) are subtracted from the fluorescence intensity (Ur) detected from the microchip (10) with the biological sample (R) fed thereto, to detect a fluorescence intensity (Uc) of fluorescence (Lk) emitted from the biological sample (R).