Abstract
A few years ago, we introduced the Centrifuge Force Microscope (CFM) as a new platform for massively parallel single-molecule manipulation. Our original prototype instrument used a microscope on a rotary stage to generate uniform centrifugal forces, which were applied to thousands of microspheres at once, hundreds of which formed single molecule tethers. Here we present a second generation CFM that is fully integrated into a commercial benchtop centrifuge, further increasing the accessibility of the method and greatly expanding the dynamic force range. We have condensed a complete video microscope to fit into a centrifuge bucket, along with battery power and an onboard computer. The result is a fully wireless “CFM module” that can be placed inside a swinging bucket, while still retaining full control of the instrument (even during centrifugation) from a wifi enabled computer. We have demonstrated the unit up to 1000g, which gives a force range of ∼70fN to 70pN using 2.8 micron microspheres. Our current device can save images at near USB 2.0 speeds with up to 5-megapixel resolution, and we are working toward improving the bandwidth further. While a number of single-molecule applications exist, one of our primary interests will be to probe the structure and function of various non-coding RNAs.
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