Micromachined Fluid Transfer Devices for Molecular Screening

描述（由申请人提供）：项目摘要/摘要：在试图增加许多高通量过程的样品密度和测定准确性时，缺乏用于1微升以下体积的快速、廉价和准确的流体转移技术正在成为限制因素。高通量实验，如化合物库重新格式化，纳升流体转移，测定小型化和分子库筛选都将大大受益于更准确和更快的流体转移技术。为了提供nL分配技术（例如金属销工具，其具有%CV> 15%的相对不精确的流体转移，或者非常昂贵且相对缓慢的声学分配）的可行替代方案，该提议提供了一种包含384个高精度微加工硅流体转移装置（FTD）的平行流体转移头的设计。已经制造了原型硅FTD阵列，其具有尖锐的硅工具，该工具设计用于刺穿微量滴定板上的箔或聚合物密封件，并在一次操作中取出样品。使用若丹明6 G在DMSO中的流体转移稀释实验来模拟化合物文库重新格式化方案，表明原型硅针工具阵列平行转移100 nL样品，%CV为2- 3%。一旦制造了硅FTD，在微成型工艺中使用硅FTD制备聚合物FTD就相对简单：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，其用于压缩成型与硅原件相同的聚合物部件。在并行使用染料转移和平板扫描仪在1至500 nL范围内对各种设计的流体转移进行初步测试后，硅和聚合物中最令人鼓舞的设计将被转移到合作者Matrical，Inc.。并在一系列实验中进行评价以确定它们在纳升流体转移和化合物库重新格式化中的功效。总之，已经设计了微机械加工的硅或聚合物工具组，其将以等于或小于任何当前技术的估计价格提供用于被动亚纳升流体转移的快得多、准确得多且更鲁棒的手段。项目叙述：本提案中开发的仪器将促进分子库筛选、化合物库重新格式化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上级性能能力的设备，预计即使是预算最有限的实验室也将能够使用这些工具。这些设备所带来的生产力和效率的提高将使人类疾病预防、检测和治疗新方法的筛选和表征取得快速进展。

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.