Application of Lab-Disk to high-throughput genomic profiling of single cells

我们开发了一种CD形的装置（Lab-disk），用于从大量细胞中简单快速地分离单个细胞。为了在该装置上进行PCR或RT-PCR，该装置由硅和玻璃制成，在本研究中不会蒸发微室中的截留溶液。经单细胞分离和PCR扩增，以200个/μl的沙门氏菌为接种量，在纸片上检测到沙门氏菌特异性invA基因的扩增。为了实现Jurkat细胞的q-PCR，通过使用Tth DNA聚合酶实现热细胞直接RT-PCR，Tth DNA聚合酶是一种热稳定聚合酶并且具有高逆转录活性。在细胞分离和装置上的连续热细胞直接RT-PCR后，在200个细胞/μl Jurkat细胞或更少的条件下检测到来自单细胞RT-PCR产物的荧光信号。

We have developed a CD-shaped device(Lab-disk) to isolate single cell from a large number of cells simply and rapidly. In order to perform PCR or RT-PCR on the device, the device was fabricated from silicon and glass not to evaporate the entrapped solution in the microchambers in this study. Amplification of Salmonella specific invA gene form single cell was confirmed on the disk at 200 cells/μl of Salmonella enterica after single cell isolation and PCR. To realize q-PCR of Jurkat cell, hot cell-direct RT-PCR was enabled by using Tth DNA Polymerase, which is a thermo-stable polymerase and has high reverse transcription activity. After cell isolation and successive hot cell-direct RT-PCR on the device, fluorescent signals from RT-PCR products for single cells were detected at 200 cells/μl of Jurkat cell or less.