Multiparameter cell sorting enables the isolation of target cells with highest purities based on the combination of different parameters, including simultaneous depletion of dead cells and debris. However, conventional droplet-based sorters with excessive pressure, high shear forces, and electrical charges can cause high stress to cells and lead to transcriptional changes, thereby introducing bias in downstream gene expression analysis. The MACSQuant® Tyto® Cell Sorter enables gentle multiparameter cell sorting minimizing cellular stress for better genomic results.
Data provided by Quy Nguyen (Kessenbrock Lab, University of California, Irvine) demonstrate lower impact on EpCAM+/CD49f+ murine mammary gland epithelial cells sorted with the MACSQuant Tyto compared to conventional cell sorters. Prior to sequencing, library prep was performed using the 10x Chromium® system.
Side-by-side comparison of gene expression of cells sorted on the MACSQuant Tyto and on a conventional droplet-based cell sorter. Dots colored in red indicate which dataset shows higher gene activation. The dot size indicates a relative gene-expression level.
Whole transcriptome (mRNA) analysis showed that cells sorted using the MACSQuant Tyto resulted in higher numbers of total detected genes and unique molecular identifiers (UMIs) compared to cells sorted on a conventional droplet sorter, indicating a high sample quality.
A matrigel sphere formation assay was performed after cells were sorted using either a droplet-based cell sorter or the MACSQuant Tyto. Seven days after seeding, the Tyto-sorted samples showed a significantly higher sphere formation, speaking to a higher viability and functionality of these cells, and matching the results observed in the scRNAseq experiments.
Unlike conventional droplet sorters, the MACSQuant Tyto Cell Sorter uses a gentle microchip-based cell sorting technology to reach highest purities and cellular viabilities without exposing cells to stressful conditions. The actual sorting process takes place within the MACSQuant Tyto Cartridge.
The cartridge provides a single-use and fully closed system, eliminating the risk of sample contamination and carry-over. In addition, due to the closed cartridge system, no aerosols will be generated during the sorting process, which is especially important for keeping operators safe when sorting infectious material. Furthermore, the instrument is equipped with a temperature-controlled cooling unit, stabilizing the samples at a temperature of choice from 4–25 °C.
Researchers know they can trust their samples in the MACSQuant Tyto and overcome puzzling applications.
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