Well Array Fluorescence Detection Using Solvent Dehydration
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Solution Overview
Problem
Existing methods for detecting target substances in miniaturized reaction spaces suffer from reduced detection sensitivity due to decreased capture of target substances, which is exacerbated by the miniaturization of reaction volumes.
Innovation Solution
A method involving a well array with first and second wells of specific volume ratios, sealed with a sealing liquid and dehydrated using a water-absorbing organic solvent to concentrate fluorescent substances for enhanced detection, combined with a fluid device and kit for target substance detection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Loss of time
If the volume of the reaction space is miniaturized, then the detection time of the enzymatic reaction is shortened, but the proportion of the target substance captured in the reaction space decreases and detection sensitivity is lowered
Solution Approach 1:
The reaction system is segmented into two distinct wells: a first well for target substance capture and a second well for enzymatic reaction. This segmentation allows the system to maintain a larger capture volume while performing reactions in a miniaturized space, thereby resolving the contradiction between detection time and detection sensitivity.
Solution Approach 2:
The second well (reaction well) is positioned at the bottom of the first well (capture well), creating a nested structure. The reaction space is defined by the volume between the sealing liquid and the bottom surface of the first well, effectively nesting the reaction chamber within the capture chamber. This nested configuration enables both capture and reaction functions within a compact structure.
2Productivity
If the volume of the reaction space is miniaturized, then the detection time is shortened, but the capture proportion of target substance decreases
Solution Approach 1:
The system separates the capture function (first well with larger volume) from the reaction function (second well with smaller volume). This allows the capture well to accumulate sufficient target substances while the reaction well maintains miniaturized dimensions for rapid detection, resolving the contradiction between detection speed and target substance capture.
Solution Approach 2:
The invention utilizes the vertical dimension by positioning the second well at the bottom of the first well. This three-dimensional arrangement allows the capture well to have a larger effective volume for target accumulation while the reaction space remains compact, achieving both high capture proportion and fast detection speed.
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 high-sensitivity detection of target substances by concentrating fluorescent signals, improving capture probability and reducing detection time.
Implementation Method 1
replacing the sealing liquid with a water-absorbing organic solvent, thereby dehydrating the content of the well, reducing the volume
Implementation Method 2
emitting excitation light to the fluorescent substance to detect fluorescent light in each well of the well array
Data Source
AI summary
A method for detecting a target substance in a sample includes a step of introducing the sample and a reagent that generates a fluorescent substance in a presence of the target substance into each well of a well array; a step of sealing each well of the well array with a sealing liquid such that each well forms an independent reaction space; a step of replacing the sealing liquid with a water-absorbing organic solvent, thereby dehydrating the content of the well, reducing the volume, and generating the fluorescent substance in a case where the target substance is present in the content; and a step of emitting excitation light to the fluorescent substance to detect fluorescent light generated in each well of the well array, in which detection of the fluorescent light in the well indicates the presence of the target substance in the well.