研究目的
To develop a DC-biased optofluidic biolaser for sensitive uric acid detection, overcoming the sensitivity limitation imposed by the laser threshold condition.
研究成果
A DC-biased optofluidic biolaser was successfully developed for sensitive UA detection, achieving a low limit of detection of 3.63 μM with a sample volume of 10 μl. This method enhances the sensitivity of optofluidic biolasers and has potential for detecting other substances in human body fluids that can be converted into H2O2.
研究不足
The sensitivity and accuracy of the sensor are dependent on the precision of the DC bias measurement. The method requires optimization of H2O2 concentration for effective bias.
1：Experimental Design and Method Selection:
A DC-biased optofluidic biolaser was developed using a Fabry-Perot laser cavity and a coupled enzyme reaction to transform UA detection into H2O2 detection. The fluorescent product resorufin served as the gain medium.
2：Sample Selection and Data Sources:
UA samples were prepared with varying concentrations, and the reaction's fluorescent product was used as the data source for detection.
3：List of Experimental Equipment and Materials:
Fabry-Perot laser cavity, microfluidic channel, 532 nm pulsed laser, spectrometer, and various reagents including uricase, HRP, and ADHP.
4：Experimental Procedures and Operational Workflow:
The enzyme reactions were carried out inside the laser cavity, with H2O2 pre-added as a DC bias. The laser emission was monitored over time to determine the UA concentration.
5：Data Analysis Methods:
The laser onset time difference was used as the sensing signal, with the integrated intensity of laser emission analyzed to determine UA concentration.
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Amplex Red Uric Acid Assay Kit
A22181
Thermo Fisher
Provides the necessary reagents for the enzyme-catalyzed reactions, including uricase and Tris-HCl reaction buffer.
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Enhanced Chemifluorescent HRP Substrate Kit
15159
Thermo Fisher
Supplies the ADHP reagent and enhancer for the enzyme-catalyzed reactions, producing the fluorescent product resorufin.
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Horseradish peroxidase (HRP)
31490
Thermo Fisher
Catalyzes the oxidation of ADHP by H2O2 to produce resorufin, the gain medium for the optofluidic laser.
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Spectrometer
HR4000
Ocean Optics
Collects and analyzes the laser emission and fluorescence signals for uric acid detection.
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Fabry-Perot laser cavity
Serves as the laser cavity for the optofluidic biolaser, enabling the detection of uric acid through enzyme-catalyzed reactions.
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532 nm pulsed laser
Continuum
Used as the pump source for the optofluidic biolaser, facilitating the lasing action within the Fabry-Perot cavity.
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