Proc. SPIE-Int. Soc. Opt. Eng. 1999, 3602 (Advances in Fluorescence Sensing Technology IV), 220-231.
Single-Analyte to Multianalyte Fluorescence Sensors
J. J. Lavigne, A. Metzger, K. Niikura, L. A. Cabell, S. M. Savoy, J. S.-J. Yoo, J. T. McDevitt, D. P. Neikirk, J. B. Shear, E. V. Anslyn
The rational design of small molecules for the selective complexation of analyltes has reached a level of sophistication such that there exists a high degree of prediction. An effective strategy for transforming these hosts into sensors involoves covalently attaching a fluorophore to the receptor which displays some fluorescence modulation when analyte is bound. Competition methods, such as those used with antibodies, are also amenable to these synthetic receptor, yet there are few examples. In our laboratories, the use of common dyes in competition assays with small molecules has proven very effective. For example, an assay for citrate in beverages and an assay for the secondary messenger IP3 in cells have been developed. Another approach we have explored focuses on multi-analyte sensor arrays which attempt to mimic the mammalian sense of taste. Our system utilizes polymer resin beads with the desired sensors covalently attached. These functionalized microspheres are then immobilized into micromachined wells on a silicon chip thereby creating our taste buds. Exposure of the resin to analyte causes a change in the transmittance of the bead. This change can be fluorescent or colorimetric. Optical interrogation of the microspheres, by illuminating from one side of the wafer and collecting the signal on the other, results in an image. These data streams are collected using a CCD camera which creates red, green, and blue (RGB) patterns that are distinct and reproducible for their environments. analysis of this data can identify and quantify the analytes present.