Wong E Y, Diamond S L
Penn Center for Molecular Discovery, Institute for Medicine and Engineering, Department of Chemical and Biomolecular Engineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA.
Anal Chem. 2009 Jan 1;81(1):509-14. doi: 10.1021/ac801959a.
In the assembly of microarrays and microarray-based chemical assays and enzymatic bioassays, most approaches use pins for contact spotting. Acoustic dispensing is a technology capable of nanoliter transfers by using acoustic energy to eject liquid sample from an open source well. Although typically used for well plate transfers, when applied to microarraying, it avoids the drawbacks of undesired physical contact with the sample; difficulty in assembling multicomponent reactions on a chip by readdressing, a rigid mode of printing that lacks patterning capabilities; and time-consuming wash steps. We demonstrated the utility of acoustic dispensing by delivering human cathepsin L in a drop-on-drop fashion into individual 50-nanoliter, prespotted reaction volumes to activate enzyme reactions at targeted positions on a microarray. We generated variable-sized spots ranging from 200 to 750 microm (and higher) and handled the transfer of fluorescent bead suspensions with increasing source well concentrations of 0.1 to 10 x 10(8) beads/mL in a linear fashion. There are no tips that can clog, and liquid dispensing CVs are generally below 5%. This platform expands the toolbox for generating analytical arrays and meets needs associated with spatially addressed assembly of multicomponent microarrays on the nanoliter scale.
在微阵列以及基于微阵列的化学分析和酶促生物分析的组装过程中,大多数方法使用针进行接触式点样。声学分配是一种能够通过利用声能从开放源孔中喷射液体样品来进行纳升转移的技术。虽然通常用于微孔板转移,但应用于微阵列时,它避免了与样品进行不必要的物理接触的缺点;通过重新寻址在芯片上组装多组分反应的困难,这是一种缺乏图案化能力的刚性打印模式;以及耗时的洗涤步骤。我们通过将人组织蛋白酶L逐滴递送至单个50纳升的预点样反应体积中,以激活微阵列上目标位置的酶反应,证明了声学分配的实用性。我们生成了范围从200到750微米(及更大)的可变大小的点,并以线性方式处理荧光珠悬浮液的转移,源孔浓度从0.1到10×10⁸个珠子/毫升增加。不存在会堵塞的尖端,液体分配的变异系数通常低于5%。该平台扩展了用于生成分析阵列的工具箱,并满足了与纳升规模的多组分微阵列的空间寻址组装相关的需求。