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Participants/Project Abstracts

Summer 2006 Bioanalytical Science REU Participants

Scott Kallgren, Hillsdale College

DEVELOPMENT OF PHOTO-PATTERNED ARRAYS FOR SPR IMAGING

Department of Chemistry, University of California - Riverside, UCR 92521 and Hillsdale College, Hillsdale, MI 49242.

Photolithography was attempted as means to produce array patterns on gold-coated glass substrates, which would subsequently be used for biodetection with surface plasmon resonance imaging (SPRi). Initial photomasks designed with MS Paint/Powerpoint and printed on transparency proved non-ideal due to minor smearing and inhomogeneity, but were nonetheless adequate. High-definition masks were designed with Adobe Photoshop and converted into photographic slides. Though these masks were found unusable due to absorption of all UV radiation, simpler slide films may still be a viable option. Fluorescence recovery after photobleaching (FRAP) experiments were employed to determine both the extent of photo-pattern removal and the extent of vesicle fusion in the wells. An inquiry into the feasibility of FRAP with PMT detection (rather than CCD) showed promise, but may require too much effort to be workable. Gold-coated slides were successfully coated with silica and subjected to photo-patterning, but etching was attempted unsuccessfully due to photoresist instability in HF. High-definition SPR images of a photo-patterned gold slide were obtained. The result is very promising for future biosesnsor array development. Low-power fluorescence microscopy of the green-to-red fluorescent photoresist indicated that there is no photoresist in the wells, and the addition of PC-NBD resulted in a blue-to-green fluorescence over the entire surface, raising the possibility that vesicles fused in the wells. It is still unclear whether the wells are sufficiently clean to be used as arrays in SPRi biodetection, but further FRAP analysis of photo-patterned chips should indicate whether vesicles fused to a biliayer in the wells. Some ambiguity could be due to chip-to-chip variation and may be corrected by using fresh photoresist, developer solution, and keeping the baking temperature more constant .

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