Cell sorting services are performed with Fluorescence-activated cell sorting (FACS cell sorting) which is a powerful tool used in both basic and clinical research that enables individual cells to be separated from a heterogeneous sample for downstream analysis or therapeutic applications. FACS sorting can be used to separate live cell populations into subpopulations based upon the fluorochrome associated with a detection antibody.
FACS cell sorting is a specialized form of flow cytometry and uses many of the same principles of cell preparation and labeling as traditional cytometry. In fact, a cell sorter works in a similar way to a flow cytometer with a single-cell suspension of fluorescently labeled cells passing through a fluidic system and interrogated by lasers that excite the fluorescent molecules used to label the different cell types. However, uniquely to cell sorters, electrostatic drop deflection, (the same principle used in ink-drop printers), is employed to target specific cell types for collection.
This is achieved by utilizing a vibration mechanism to create break points in the fluidics stream that result in the generation of distinct droplets. By fine-tuning the frequency of the vibrations, the frequency of the break points can be optimized to generate droplets containing single cells. Charge (hi/low positive or negative) is applied to droplets containing cells of interest, enabling them to be diverted into collection tubes using electrostatically charged deflection plates.
This enables relatively pure cell populations to be collected. Typically, 2 to 4 different cell populations can be sorted simultaneously from a single heterogeneous population. Populations can be sorted based on a variety of characteristics – essentially, if you can differentiate a cell, you can sort it.
- TBNK (CD16, CD56, CD19, CD25, CD14, CD4, CD8, CD3, HLA-DR, CD45)
- T subsets (CD45RA, CD197/CCR7, CD28, CD279/PD1, CD27, CD4, CD8, CD3, CD57, CD45)
- T reg intranuclear (CD45RA, CD25, CD39, CD4, FoxP3, CD3, Helios, CD45)
- TCR (TCR gamma-delta, TCR alpha-beta, HLA-DR, TCR Vdelta1, CD4, CD8, CD3, TCR Vdelta2, CD45)
- B cells (IgD, CD21, CD19, CD27, CD24, CD38, IgM, CD45)
- DCs (CD16, LINEAGE, CD1c, CD11c, Clee9A, CD123, HLA-DR, CD45)
- Intracellular (IFN gamma, TNF alpha, IL-2, CD8, CD3, CD4)
- DTC panel 1 (CD16, CD56, CD19, CD25, CD39, CD14, CD4, CD8, CD3, HLA-A2, CD45)
- DTC panel 2 (CD47, CD133, CD326, 7AAD, CD324, CD274, CD44, HLA-A2, CD45)
- ALL (CD7, CD79b, CD117, CD19, CD10, CD4, CD8, CD3, CD5, CD45)
- CLL (KAPPA, LAMBDA, CD117, CD19, CD10, CD20, CD23, CD43, CD5, CD45)
- MM1 (KAPPA, LAMBDA, CD48, CD56, CD117, CD138, CD38, CD33, CD19, CD45)
- MM2 (CD9, CD13, CD10, CD3, CD117, CD138, CD38, CD33, CD19, CD45)
- Histone DeAc (fixable L/D, H3 K27Ac, CD19, CD56, CD114, CD4, CD8, CD3, HLA-DR,DP,DQ, CD45)