研究目的
To develop a method for creating microfluidic oxygen sensors that enable simultaneous imaging of spatial structure and chemical oxygen gradients using solvent-induced fluorophore impregnation (SIFI) in polystyrene devices.
研究成果
The SIFI method successfully creates oxygen-sensitive surfaces in polystyrene microfluidic devices, enabling simultaneous imaging of spatial structure and oxygen gradients. The approach is reproducible and generalizable to other polystyrene containers for sensing applications, with potential uses in cell culture and biomedical devices.
研究不足
The technique is limited to polystyrene materials; diffusion of oxygen through thin polystyrene devices might bias measurements in slow-flow or respiratory processes, requiring anoxic chambers or hybrid devices with oxygen-impermeable layers. The luminescence decay under air is multiexponential, complicating calibration.
1:Experimental Design and Method Selection:
The SIFI method was used to impregnate interior surfaces of polystyrene microfluidic structures with PtTFPP dye. This involved exposing the surfaces to acetonitrile solutions of the dye at controlled temperatures to create a gel layer for dye diffusion.
2:Sample Selection and Data Sources:
Polystyrene sheet material and fabricated microfluidic devices (straight microchannel and pore network devices) were used. Dyes included NPO for development and PtTFPP for oxygen sensing.
3:List of Experimental Equipment and Materials:
Equipment included an AML wafer bonder for bonding, Etaluma Lumascope 600 for cross-sectional imaging, Lambert LIFA-P system for fluorescence lifetime imaging, Leica DMI6000 microscope, mass flow controllers for gas mixing, milliGAT pump for liquid flow, and Micromodule G591 for dissolved oxygen equilibration. Materials included polystyrene sheets, PtTFPP and NPO dyes, acetonitrile solvent, and bonding components.
4:Experimental Procedures and Operational Workflow:
Devices were fabricated by hot embossing and bonding. Impregnation involved injecting dye solutions into devices at 50°C for specific times, followed by drying. Imaging was performed using fluorescence microscopy and FLIM under controlled oxygen conditions. Calibrations were done with gaseous and dissolved oxygen samples.
5:Data Analysis Methods:
Fluorescence lifetime data were analyzed using frequency domain techniques, with phasor plots and Stern-Volmer representations for calibration. Statistical variations were considered from pixel-to-pixel data.
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Microscope
DMI6000
Leica
Microscope platform for mounting the Lambert LIFA-P system for imaging.
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LIFA-P
LI2CAM-P
Lambert Instruments
Fluorescence lifetime imaging system for measuring oxygen concentrations via frequency domain techniques.
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Lumascope
600
Etaluma
Inverted microscope for cross-sectional bright field and fluorescence imaging.
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Wafer Bonder
AML
Used for bonding polystyrene sheets in microfluidic device fabrication.
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Mass Flow Controller
Controls gas flows for preparing gaseous oxygen mixtures for calibration.
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Pump
milliGAT
Global FIA
Pumps water flow for dissolved oxygen experiments.
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Membrane Device
G591
Liqui-Cel
Equilibrates liquid with gas for dissolved oxygen control using hydrophobic polypropylene hollow fiber membranes.
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UV Light
UVGL-58
Mineralight
Conventional UV light source for imaging overall structures.
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CCD Camera
DSC-TX20
Sony
Camera for capturing images under UV light.
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