A 2-D microcantilever array for multiplexed biomolecular analysis

An accurate, rapid, and quantitative method for analyzing variety of biomolecules, such as DNA and proteins, is necessary in many biomedical applications and could help address several scientific issues in molecular biology. Recent experiments have shown that when specific biological reactions occur on one surface of a microcantilever beam, the resulting changes in surface stress deflect the cantilever beam. To exploit this phenomenon for high-throughput label-free biomolecular analysis, we have developed a chip containing a two-dimensional (2-D) array of silicon nitride cantilevers with a thin gold coating on one surface. Integration of microfluid cells on the chip allows for individual functionalization of each cantilever of the array, which is designed to respond specifically to a target analyte. An optical system to readout deflections of multiple cantilevers was also developed. The cantilevers exhibited thermomechanical sensitivity with a standard deviation of seven percent, and were found to fall into two categories-those whose deflections tracked each other in response to external stimuli, and those whose did not due to drift. The best performance of two "tracking" cantilevers showed a maximum difference of 4 nm in their deflections. Although "nontracking" cantilevers exhibited large differences in their drift behavior, an upper bound of their time-dependent drift was determined, which could allow for rapid bioassays. Using the differential deflection signal between tracking cantilevers, immobilization of 25mer thiolated single-stranded DNA (ssDNA) on gold surfaces produced repeatable deflections of 80 nm or so on 0.5-μm-thick and 200-μm-long cantilevers.