Micromachined Fluid Transfer Devices for Molecular Screening

描述（由申请人提供）：项目摘要/摘要：缺乏快速、廉价和准确的流体传输技术，用于1微升以下的体积，这正在成为许多高通量过程中试图提高样品密度和分析准确性的限制因素。高通量实验，如化合物文库重组、纳升流体转移、实验小型化和分子文库筛选，都将从更准确、更快速的流体转移技术中受益匪浅。为了提供一种可行的替代nL点胶技术，如金属销工具，其流体传输相对不精确，% cv为bb0 - 15%，或非常昂贵且相对缓慢的声学点胶，该提案提供了一种包含384个高精度微机械硅流体传输装置（FTDs）的并行流体传输头的设计。原型硅FTD阵列已经被制造出来，带有尖锐的硅工具，设计用于刺穿微量滴定板上的箔或聚合物密封，并在一次操作中提取样品。在DMSO中使用罗丹明6G进行流体转移稀释实验，模拟化合物库重组方案，结果表明，原型硅针工具阵列以2-3%的% cv平行转移100nL样品。一旦硅FTD被制造出来，在微成型工艺中使用硅FTD制备聚合物FTD就相对简单了：硅FTD用于制备正极硅母模，镍钴合金在其上电沉积（即电成型），以产生金属负极，用于压缩模制与硅原件相同的聚合物部件。在使用染料转移和平板扫描仪对各种设计进行1至500 nL范围内的流体转移初步测试后，最令人鼓舞的硅和聚合物设计将转移给合作伙伴Matrical， Inc.，并在一系列实验中进行评估，以确定它们在纳升流体转移和化合物库重新格式化方面的功效。总之，微机械硅或聚合物工具集的设计将为被动亚纳升流体输送提供更快、更准确和更强大的手段，其估计价格等于或低于任何现有技术。项目描述：本提案中开发的仪器将促进分子文库筛选，化合物文库重组和任何其他需要处理小体积液体的方法。通过以比现有技术更低的价格制造具有卓越性能的设备，预计即使是预算最有限的实验室也将获得这些工具。这些设备使生产力和效率的提高成为可能，这将使人类疾病预防、检测和治疗的新方法的筛选和表征方面取得迅速进展。

DESCRIPTION (provided by applicant): Project Summary/Abstract: The lack of rapid, inexpensive and accurate fluid transfer technology for volumes below 1 microliter is becoming a limiting factor in attempts to increase the sample density and assay accuracy of many high throughput processes. High throughput experiments such as compound library reformatting, nanoliter fluid transfer, assay miniaturization and molecular library screening would all greatly benefit from a more accurate and faster fluid transfer technology. To provide a viable alternative to nL dispensing technology such as metal pin tools, which have relatively imprecise fluid transfers with %CVs of >15%, or very expensive and relatively slow acoustic dispensing, this proposal proffers a design for a parallel fluid transfer head containing 384 highly precise micromachined silicon fluid transfer devices (FTDs). Prototype silicon FTD arrays, with sharp, pointed silicon tools designed to puncture foil or polymer seals on a microtiter plate and withdraw the sample in a single operation, have been fabricated. Fluid transfer dilution experiments using Rhodamine 6G in DMSO to simulate compound library reformatting protocols, show that the prototype silicon pin tool array transfers 100nL samples in parallel with %CVs of 2-3%. Once the silicon FTDs are fabricated, it is relatively straightforward to prepare a polymer FTD employing the silicon FTD in a micromolding process: the silicon FTDs are used to prepare a positive silicon master mold onto which a nickel-cobalt alloy is electrodeposited (i.e. electroformed) to create a metal negative which is used to compression mold a polymer part identical to the silicon original. After initial testing of fluid transfer in the 1 to 500 nL range in a variety of designs at Parallel using dye transfer and a plate scanner, the most encouraging designs in both silicon and polymer will be transferred to collaborator Matrical, Inc. and evaluated in a series of experiments to determine their efficacy in nanoliter fluid transfer and compound library reformatting. In summary, micromachined silicon or polymers tool sets have been designed which will provide a much faster, accurate and more robust means for passive sub-nanoliter fluid transfer at an estimated price equal to or less than any of the current technologies. Project Narrative: The instruments developed in this proposal will facilitate molecular library screening, compound library reformatting and any other methodology that requires handling small volumes of liquid. By making devices that possess superior performance capabilities at lower prices than existing technologies, it is anticipated that even the most modestly budgeted of laboratories will have access to these tools. The increases in productivity and efficiency made possible by these devices will enable rapid progress in the screening and characterization of new methods for the prevention, detection, and treatment of human disease.