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Microfluidic Sensors with Impregnated Fluorophores for Simultaneous Imaging of Spatial Structure and Chemical Oxygen Gradients
journal contribution
posted on 2019-01-04, 00:00 authored by Jay W. Grate, Bingwen Liu, Ryan T. Kelly, Norman C. Anheier, Thomas M. SchmidtInterior
surfaces of polystyrene microfluidic structures were impregnated
with the oxygen sensing dye Pt(II) tetra(pentafluorophenyl)porphyrin
(PtTFPP) using a solvent-induced fluorophore impregnation (SIFI) method.
Using this technique, microfluidic oxygen sensors are obtained that
enable simultaneous imaging of both chemical oxygen gradients and
the physical structure of the microfluidic interior. A gentle method
of fluorophore impregnation using acetonitrile solutions of PtTFPP
at 50 °C was developed leading to a 10-μm-deep region containing
fluorophore. This region is localized at the surface to sense oxygen
in the interior fluid during use. Regions of the device that do not
contact the interior fluid pathways lack fluorophores and are dark
in fluorescent imaging. The technique was demonstrated on straight
microchannel and pore network devices, the latter having pillars of
300 μm diameter spaced center to center at 340 μm providing
pore throats of 40 μm. Sensing within channels or pores and
imaging across the pore network devices were performed using a Lambert
LIFA-P frequency domain fluorescence lifetime imaging system on a
Leica microscope platform. Calibrations of different devices prepared
by the SIFI method were indistinguishable. Gradient imaging showed
fluorescent regions corresponding to the fluid pore network, dark
pillars, and fluorescent lifetime varying across the gradient, thus
providing both physical and chemical imaging. More generally, the
SIFI technique can impregnate the interior surfaces of other polystyrene
containers, such as cuvettes or cell and tissue culture containers,
to enable sensing of interior conditions.
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300 μ m diameterMicrofluidic Sensorspore throatspolystyrene containers10-μ m-deep region340 μ msense oxygenchemical oxygen gradientsfluid pathways lack fluorophoresLeica microscope platformchemical imaging40 μ msolvent-induced fluorophore impregnationSimultaneous Imagingmicrofluidic oxygen sensorsLambert LIFA-P frequency domain fluorescence lifetime imaging systemChemical Oxygen Gradients Interior surfacesSIFI methodpore network devicesfluid pore networkSpatial Structurepolystyrene microfluidic structuresacetonitrile solutionstissue culture containersImpregnated Fluorophoresfluorophore impregnationGradient imagingSIFI technique
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