Micromachined Fluid Transfer Devices for Molecular Screening

描述(由申请人提供)：项目摘要/摘要：1微升以下体积的快速、廉价和准确的流体传输技术的缺乏正在成为许多高通量过程提高样品密度和分析准确性的限制因素。高通量实验，如化合物文库重排、纳升流体传输、检测小型化和分子文库筛选，都将大大受益于更准确和更快的流体传输技术。为了提供一种可行的替代NL分配技术的方法，例如金属销工具，其具有相对不精确的流体传输(%CV为15%)，或者非常昂贵且相对较慢的声学分配，该提议提供了一种包含384个高精度微机械硅流体传输装置(FTD)的并行流体传输头的设计。已经制造出了原型硅FTD阵列，它带有锋利的尖头硅工具，设计用于在微滴定板上刺穿箔或聚合物密封件，并在一次操作中提取样品。用罗丹明6G在DMSO中进行的流体传输稀释实验模拟了化合物库的重新格式化协议，结果表明，原型硅针工具阵列以2-3%的%CVs并行传输100nL的样品。一旦制造了硅FTD，在微模制过程中使用硅FTD来制备聚合物FTD相对简单：硅FTD用于制备正硅母模，在其上电沉积(即电铸)镍钴合金以产生金属负极，该金属负极用于对与硅原件相同的聚合物部件进行模压。在使用染料转印和平板扫描仪并行测试了各种设计中1到500 nL范围内的流体转印后，硅和聚合物中最鼓舞人心的设计将被转给合作伙伴Matric，Inc.，并在一系列实验中进行评估，以确定它们在纳升流体转印和化合物库重新格式化方面的有效性。总而言之，微机械加工的硅或聚合物工具组的设计将为被动亚纳升流体传输提供更快、更准确和更坚固的手段，估计价格等于或低于任何当前技术。项目简介：本提案中开发的仪器将促进分子文库筛选、化合物文库重新格式化和任何其他需要处理少量液体的方法。通过以比现有技术更低的价格制造具有卓越性能的设备，预计即使是预算最低的实验室也可以使用这些工具。这些设备带来的生产率和效率的提高将使人类疾病的预防、检测和治疗的新方法的筛选和表征方面取得快速进展。

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.