A robotics platform for automated batch fabrication of high density, microfluidics-based DNA microarrays, with applications to single cell, multiplex assays of secreted proteins

Abstract

Microfluidics flow-patterning has been utilized for the construction of chip-scale miniaturized DNA and protein barcode arrays. Such arrays have been used for specific clinical and fundamental investigations in which many proteins are assayed from single cells or other small sample sizes. However, flow-patterned arrays are hand-prepared, and so are impractical for broad applications. We describe an integrated robotics/microfluidics platform for the automated preparation of such arrays, and we apply it to the batch fabrication of up to eighteen chips of flow-patterned DNA barcodes. The resulting substrates are comparable in quality with hand-made arrays and exhibit excellent substrate-to-substrate consistency. We demonstrate the utility and reproducibility of robotics-patterned barcodes by utilizing two flow-patterned chips for highly parallel assays of a panel of secreted proteins from single macrophage cells.

Flow-pattern Guided Fabrication of High-density Barcode Antibody Microarray

“…For instance, to create a 5 x 5 array, anchor sequences A to E may be used, while bridging sequences A'-i to E'-xxv are utilized. To automate the flow patterning process, a robotics platform has been developed 18 although this has only utilized one DNA flow patterning step. In principle, the same platform can be extended to the second step of perpendicular flow patterning, while the switching between two steps may require manually peeling off the first PDMS mold after the first patterning step, followed by washing and drying the slide (this step would take about 10 min), before mating the slide with another PDMS mold to facilitate the perpendicularly oriented second patterning step.…”

Biological Laser Printing Technology and Its Applications

“…吸收层上形成深 20~100 μm 的吸收小孔来控制 [8,18] ， 如图 4 所示， 每个孔的尺寸大约为 0. [25] ， 是发展基因组学、 蛋白质组学、 基因表达研究和生物医学诊 断 的 基 本 工 具 [26][27][28][29] [30][31][32][33] 。这些方法因简单易行已较为成熟， 但也存在一些局限性， 如打印后生物分子活性不高及空间分辨不…”

Microfluidics-Based Single-Cell Functional Proteomics for Fundamental and Applied Biomedical Applications

“…Of these choices, stamping does not permit the required level of multiplexing, whereas dip pen does not yield a surface coverage sufficient for stable and sensitive assays. Thus, we have developed microfluidic www.annualreviews.org • Single-Cell Functional Proteomicsflow patterning into the method of choice for SCBCs, including even building robotics systems to automate the task (79). In Figure 4 we provide data showing the influence of various barcoding surface chemistries on assay sensitivity, some of which are described in Reference 20.…”