Cells are cultured on a flexible silicon substrate, and treated with a detergent. This treatment dissolves the majority of the cell, but leaves behind the bare cytoskeleton proteins which remain bound to the substrate. The silicon can be stretched or compressed to directly apply force to the cytoskeletal structures. This allows for rapid in vitro analysis of these structures under a force load. For example, the binding of labeled proteins can be monitored to determine the effect of contraction or stretching on binding. Furthermore, this technique can be conducted in the presence of potential drugs, to quickly screen for their effect and potency in modulating tissue function or stiffness. This application is highly sought after for the development of engineered tissues, as well as novel treatments for various injuries and tissue damage.
Other methods to apply force to proteins, such as AFM or optical tweezers, are limited by the difficulty of setting up experiments and the time to complete an analysis. Additionally, these instruments are very expensive and difficult to operate and maintain. This method allows for multiple measurements to be conducted at the same time, greatly increasing the rate of experiments that can be conducted by a single researcher. The speed of analysis will be greatly useful for high-throughput screening of small molecules to assess their medical use in tissue engineering and injury treatment.
This method was verified by correctly identifying signaling molecules that bind only to force strained cytoskeletal proteins.
Patent Issued (US 6,982,150)
Available for licensing and sponsored research support
Tech Ventures Reference: IR M02-045
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