NER: Nanoengineered Pipettes for Patch Clamp Measurements

斯坦福大学richard Zare生物学的基本过程发生在纳米尺度上。蛋白质只有几纳米大小；细胞功能所必需的细胞器有几十或几百纳米。对这些结构的直接研究由于难以制造出兼容尺寸的工具而受到限制。提出了一种新的工具，可以使操纵和研究单个纳米尺度生物对象具有现有工具无法实现的控制和灵活性。一种生产移液器的纳米工程工艺已经被设计用于对小生物物体（如囊泡）的稳健操作。与电生理学中常规使用的传统移液器不同，所提出的移液器在其尖端有一个凹的座位表面。这个座面提供了一个大的接触面积之间的移液器和被持有的对象，从而产生良好的稳定性和控制。此外，座椅表面的尺寸可以定制，以容纳不同尺寸的物体，从几十微米到几百纳米不等。较低的尺寸限制与广泛的亚细胞细胞器兼容。移液器的另一个扩展是作为输送装置。提出了将膜片钳和毛细管电泳与激光诱导荧光相结合的电生理和化学成分分析方法。通过结合形态学、电生理和化学成分的信息，这种新的分析技术耦合将能够进一步阐明囊泡与囊泡之间的差异。生物异质性的重要性才刚刚开始被探索。我们提出的工具可能为这个有趣的生物学问题提供新的见解。

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.