✍🏼 Nicole Jagnandan, Jose Morachis

🏠 NanoCellect Biomedical, San Diego, CA

📑 Biomicrofluidics (2022)

Abstract

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.

How is the WOLF used in this study
the authors describe how the WOLF Cell Sorter was integrated into workflows to improve genomic sample quality, particularly for single-cell RNA-sequencing applications. The WOLF sorter uses a microfluidic, low-pressure sorting technology to gently remove debris, dead cells, and unwanted populations from heterogeneous samples, which helps preserve cell viability and RNA integrity prior to cDNA library preparation. By sorting cells with multi-parameter fluorescence gating while minimizing shear stress and sorter-induced cellular stress seen with traditional droplet sorters, the WOLF enabled cleaner, more viable cell populations that generated higher-quality sequencing data, including increased genes detected per cell and improved library complexity. Its gentle sorting mechanism makes it especially useful for sensitive downstream genomic assays where maintaining the native state of cells is critical.