MACSQuant® Tyto® cell sorting applications

Cell sorting applications on the MACSQuant® Tyto® Sorter

Sorting of immune cells

Sorting of antigen-specific T cells to high purity using the MACSQuant Tyto Sorter.
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Sterile sorting of antigen-specific T cells for clinical applications

In this FDA-approved clinical phase II study, peptide-MHC-multimers were used to detect and sort a rare subset of antigen-specific T cells out of PBMCs using the MACSQuant Tyto Sorter. The closed cartridge system facilitates contamination-free cultivation and expansion of these cells after sorting.

Sorting of pre-enriched regulatory T cells to high purity using the MACSQuant Tyto Sorter.
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Sterile sorting of pre-enriched regulatory T cells for medical applications

In this study, the CliniMACS Prodigy® was used for immunomagnetic and fluorescent labeling and cell isolation in order to pre-enrich white blood cells obtained via leukapheresis.

Subsequently, cells were sorted on the MACSQuant Tyto. Combining these closed-system instruments guarantees full sterility of your medical product.  
 

Subsequent sorting of PBMC-derived B cells, T cells, and NK cells to high purity using the MACSQuant Tyto Sorter.
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Subsequent sorting of B cells, T cells, and NK cells from one population of peripheral blood mononuclear cells (PBMCs)

Unlike conventional droplet sorters, sorting on the MACSQuant Tyto allows for retrieval of the negative population and permits subsequent sorting of functionally different cell populations out of one single population of cells. In this study, B cells, T cells and NK cells were subsequently sorted out of one population of PBMCs without affecting cell viability.

Sorting of neutrophils to high purity using the MACSQuant Tyto Sorter.
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Gentle sorting of neutrophils

Neutrophil granulocytes are very sensitive towards environmental changes like mechanical stress, temperature changes, and other factors. Therefore, gentle isolation is a critical step for accurate analysis of their function.

Here, we sorted neutrophil granulocytes out of non-lysed whole blood to high purity using the MACSQuant Tyto Sorter. No difference in expression of activation and naïve markers could be observed compared to other
separation methods. 

Sorting of different NK cell populations from one starting material using the MACSQuant Tyto Sorter.
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Gentle subsequent sorting of different NK cell populations

Due to the unique cartridge design, the MACSQuant Tyto Sorter allows for a completely sterile recovery of the negative fraction after the sort process. In addition to  the prevailing gentle sorting conditions, this permits subsequent sorts from one and the same starting sample.

In this study, the two major subtypes of NK cells present in human blood were subsequently sorted out of one population of peripheral blood mononuclear cells (PBMCs) without affecting cell viability.



Sorting of stem cells 

Sorting of iPSCs to high purity using the MACSQuant Tyto Sorter.
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Sorting of induced pluripotent stem cells (iPSCs) with preserved functionality

GFP-positive IPSCs were sorted using the MACSQuant Tyto, then replated and cultured for 24 hours. When compared to controls, GFP-positive IPSCs showed comparable replating efficiencies, highlighting the gentle nature of sorting with the MACSQuant Tyto. Subsequently these cells where differentiated into cardiomyocytes and sorted to 98% purity. After replating, cardiomyocytes started contracting within 24–48 hours, confirming functionality of cells.


Sorting of mesenchymal stem cells to high purity using the MACSQuant Tyto Sorter.
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Gentle sorting of rare mesenchymal stem cells

Due to the gentle sorting conditions and because all cells remain in the cartridge after each sort, sequential sorting can be applied to enrich rare cells in a two-step sorting strategy. In this study bone marrow derived mesenchymal stem cells were sorted to high purity in a sequential sorting strategy using the MACSQuant Tyto. 

Sorting of CD133+ cells to high purity using the MACSQuant Tyto Sorter.
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Gentle sorting of CD133+ cells from frozen cord blood mononuclear cells (MNCs)

Gentle sorting conditions and a completely sterile cartridge system, in which cells remain at all times, allow for efficient enrichment of rare cells on the MACSQuant Tyto Sorter, e.g., by using a two-step sorting strategy.

In this experiment, rare CD133+ cells were sorted from frozen cord blood MNCs. The positive fraction of the first sort was used as input for a second sort, yielding a highly pure cell population.



Sorting of high-risk material

Sorting of yeast cells to high purity using the MACSQuant Tyto Sorter.
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Completely safe sorting of yeast cells

Due to the closed cartridge system, the MACSQuant Tyto allows for completely safe sorting of hazardous cell material, such as bacteria, yeast or potentially contaminated cells. In this experiment, GFP-positive yeast cells were sonicated and sorted to high purity without affecting viability. On the MACSQuant Tyto Sorter subsequent experiments can directly be executed without a labor-intensive cleaning routine or risk for persistent contamination of your sorter. 

Sorting of GFP+ bacteria to high purity using the MACSQuant Tyto Sorter.
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Sorting of bacteria without any risk

Within the closed MACSQuant Tyto Cartridge, the formation of biohazardous aerosols or droplets is completely eliminated, making it a completely safe system. This is particularly important with regards to operator safety when working with high-risk material, such as bacteria. In addition, the closed cartridge design excludes problems like sample-to-sample carry-over or the risk of persistent instrument contamination. Due to the single-use disposable cartridge, there is no need for difficult sterilization procedures between subsequent sorts

This experiment provides a proof of principal for safe sorting of very small (E. coli diameter ≈ 1µm, length ≈ 2µm), high-risk material using the MACSQuant Tyto Sorter. GFP-expressing bacteria (E. coli) were sorted from a mixture with wildtype bacteria, resulting in a sort purity of 97%.



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