Fiber-optic chemical sensor microarrays for the detection of pH and O2 have been developed with subsecond response times. Sensor microarrays are fabricated by the covalent immobilization (pH sensor arrays) or the physical entrapment (O2 sensor arrays) of fluorescent indicators in photodeposited polymer matrices on optical imaging fibers. Polymer microarrays are comprised of thousands of individual elements photodeposited as hemispheres such that each element of the sensor array is coupled directly to a discrete optical element of the imaging fiber and is not in contact with other neighboring elements. Because of the hemispherical shape and the individuality of the array elements, diffusion of analyte to the sensor elements is dominated by radial diffusion, resulting in a rapid response time. pH-sensitive arrays based on fluorescein respond to a 1.5-unit pH change within 300 ms, while the O2-sensitive arrays respond to O2 changes within 200 ms (90% of steady state response).
An optical penicillin biosensor is described, based on the enzyme penicillinase. The sensor is fabricated by selective photodeposition of analyte-sensitive polymer matrices on optical imaging fibers. The penicillin-sensitive matrices are fabricated by immobilizing the enzyme as micrometer-sized particles in a polymer hydrogel with a covalently bound pH indicator. An array of penicillin-sensitive and pH-sensitive matrices are fabricated on the same fiber. This array allows for the simultaneous, independent measurement of pH and penicillin. Independent measurement of the two analytes allows penicillin to be quantitated in the presence of a concurrent pH change. An analysis was conducted of enzyme kinetic parameters in order to model the penicillin response of the sensor at all pH values. This analysis accounts for the varying activity of the immobilized penicillinase at different pH values. The sensor detects penicillin in the range 0.25-10.0 mM in the pH range 6.2-7.5. The sensor was used to quantify penicillin concentration produced during a Penicillium chrysogenum fermentation.
Microstructures were fabricated on optical imaging fibers with a photopolymerization technique. Monodisperse polymeric microarrays were produced containing spots of 2.5 micrometers in diameter spaced 4.5 micrometers apart. Polymer microarrays were also deposited on other substrates by using imaging fibers for light delivery. The technique allows micrometer-scale photopatterning with masks larger than the desired dimensions.
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