研究目的
To develop a portable lab-on-chip system for real-time monitoring of enzymatic chemiluminescent reactions, specifically for detecting hydrogen peroxide using horseradish peroxidase as a model enzyme.
研究成果
The developed portable lab-on-chip system successfully integrates microfluidics and optoelectronics for real-time monitoring of chemiluminescent enzymatic reactions, achieving a low limit of detection for hydrogen peroxide and demonstrating feasibility for biosensing applications, with potential for future expansions to other analytes and improvements in sensitivity.
研究不足
The system may have constraints in terms of the specific enzymes and analytes tested (e.g., limited to horseradish peroxidase and hydrogen peroxide), potential issues with reusability and long-term stability, and the need for further optimization for broader applications.
1：Experimental Design and Method Selection:
The system integrates a microfluidic network with amorphous silicon photosensors for on-chip chemiluminescence detection, designed to reduce optical losses and enhance compactness.
2：Sample Selection and Data Sources:
Horseradish peroxidase is immobilized on microchannel walls as the enzyme model; hydrogen peroxide solutions at various concentrations are used as analytes.
3：List of Experimental Equipment and Materials:
Glass substrates (Borofloat 33), hydrofluoric acid for etching, amorphous silicon layers deposited via PECVD, electronic boards for photocurrent read-out, metallic box for shielding, and reagents like luminol and 4-iodophenol.
4：Experimental Procedures and Operational Workflow:
Fabrication involves wet etching of microchannels, anodic bonding of glass substrates, deposition and patterning of photosensors, functionalization of channels with polymer brushes and enzyme immobilization, followed by flow of reagent mixtures and photocurrent measurement.
5：Data Analysis Methods:
Photocurrents are read out and analyzed to determine limits of detection and quantification, with comparisons to literature values.
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Borofloat 33
Used as the glass substrate for microfluidic network fabrication.
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Hydrofluoric acid
25%
Used for wet etching of the glass substrate to define microfluidic channels.
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Amorphous silicon photosensors
p-i-n junction
Detect chemiluminescent radiation and produce photocurrents proportional to light emission.
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Plasma Enhanced Chemical Vapor Deposition system
Used for deposition of amorphous silicon layers.
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SU-8 3005
3005
Used as an insulation layer in the fabrication process.
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Indium Tin Oxide
Used as the bottom contact for the photosensors.
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Titanium Tungsten
TiW
Used as the top contact for the photosensors.
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Metallic box
Encloses the system to provide shielding from electromagnetic interferences.
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Te?on-holder
Ensures sealed connection to external tubes for reagent flow.
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Electronic boards
Read out the photocurrents from the photosensors.
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