NER: Nanoengineered Pipettes for Patch Clamp Measurements

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AbstractCTS-0102889Richard Zare, Stanford UniversityThe fundamental processes of biology occur on the nanoscale. Proteins are on the order of few nanometers; the organelles essential to cellular function are tens or hundreds of nanometers. Direct investigation of these structures has been limited by the difficulty of making tools of compatible size. A novel tool is proposed that may enable the manipulation and investigation of individual nanoscale biological objects with control and flexibility impossible with existing tools. A nanoengineering process that produces pipettes especially has been designed for the robust manipulation of small biological objects, such as vesicles. Unlike the conventional pipettes routinely used in electrophysiology, the proposed pipettes have a concave seating surface at their tips. This seating surface provides a large contact area between the pipette and the object being held, resulting in excellent stability and control. In addition, the size of the seating surface can be tailored to hold objects of varying sizes, from tens of microns to hundreds of nanometers. The lower size limit is compatible with a wide range of subcellular organelles.A further extension of our pipettes is as delivery devices. It is proposed to combine electrophysiological and chemical composition analyses by coupling patch clamp and capillary electrophoresis with laser induced fluorescence. This novel coupling of analysis techniques will enable the further elucidation of vesicle-to-vesicle differences by combining information about morphology, electrophysiology, and chemical composition. The significance of biological heterogeneity is only beginning to be explored. The tools we propose may afford new insights on this intriguing biological problem.