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

我们开发了一种CD形状的装置(Lab-Disk)，可以简单、快速地从大量细胞中分离单个细胞。为了对该装置进行聚合酶链式反应或逆转录-聚合酶链式反应，在本研究中，该装置是由硅和玻璃制成的，不会蒸发微室中的包埋溶液。经单细胞分离和聚合酶链式反应，证实了沙门氏菌特异性invA基因从单细胞中扩增，扩增速度为200cell/μ。为了实现Jurkat细胞的Q-PCR，利用TTH DNA聚合酶实现了热细胞直接RT-PCR，TTH DNA聚合酶是一种具有很高反转录活性的热稳定性聚合酶。在该装置上进行细胞分离和连续热细胞直接RT-PCR后，从单个细胞的RT-PCR产物中可以检测到200cell/μ或以下的Jurkat细胞的荧光信号。

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.