Jagnandan Nicole, Morachis Jose
Applications, NanoCellect Biomedical Inc., San Diego, California 92121, USA.
Biomicrofluidics. 2022 Jun 10;16(3):034106. doi: 10.1063/5.0092358. eCollection 2022 May.
Single-cell RNA-Sequencing has led to many novel discoveries such as the detection of rare cell populations, microbial populations, and cancer mutations. The quality of single-cell transcriptomics relies heavily on sample preparation and cell sorting techniques that best preserve RNA quality while removing dead cells or debris prior to cDNA generation and library preparation. Magnetic bead cell enrichment is a simple process of cleaning up a sample but can only separate on a single-criterion. Droplet-based cell sorters, on the other hand, allows for higher purity of sorted cells gated on several fluorescent and scatter properties. The downside of traditional droplet-based sorters is their operational complexity, accessibility, and potential stress on cells due to their high-pressure pumps. The WOLF Cell Sorter, and WOLF G2, developed by NanoCellect Biomedical, are novel microfluidic-based cell sorters that use gentle sorting technology compatible with several RNA-sequencing platforms. The experiments highlighted here demonstrate how microfluidic sorting can be successfully used to remove debris and unwanted cells prior to genomic sample preparation resulting in more data per cell and improved library complexity.
单细胞RNA测序带来了许多新发现,例如检测稀有细胞群体、微生物群体和癌症突变。单细胞转录组学的质量在很大程度上依赖于样本制备和细胞分选技术,这些技术能在生成cDNA和制备文库之前,最好地保存RNA质量,同时去除死细胞或碎片。磁珠细胞富集是一种简单的样本清理过程,但只能基于单一标准进行分离。另一方面,基于微滴的细胞分选仪能够根据多种荧光和散射特性,实现更高纯度的分选细胞。传统基于微滴的分选仪的缺点在于其操作复杂性、可及性,以及由于高压泵对细胞造成的潜在压力。由NanoCellect Biomedical公司开发的WOLF细胞分选仪和WOLF G2,是新型的基于微流控的细胞分选仪,它们采用了与多种RNA测序平台兼容的温和分选技术。此处重点介绍的实验展示了微流控分选如何能在基因组样本制备之前成功用于去除碎片和不需要的细胞,从而使每个细胞获得更多数据并提高文库复杂性。