Want to purify immune cell populations for downstream assays? Sorted cells can be used for genetic analyses, proteomics studies, protein expression (by western blot), cellular assays (cytokine secretion, proliferation, killing assays, drug treatment assays, developmental progression, differentiation with individual culture conditions, and more). Cells can also be sorted to separate different populations of genetically-modified cells (transfections, RNAi knockdown, CRISPR targeted cells). We can prepare your sample, stain with the antibodies of interest, and sort specific cell populations. We can also run your pre-stained samples on our instruments. At HIMSR, we offer full service cell sorting. Log into iLabs for service fees; contact Kim Jordan for a quote.
Fluorescence-Activated Cell Sorting (FACs)
We use the BD FACSAria Fusion, a flow cytometry-based cell sorter capable of separating individual cells based on light scatter properties (forward/side scatter), membrane permeability (live/dead exclusion), and expression of up to 12 surface markers. With this instrument, you can separate up to 4 different cell populations at once, and drop individual cells into 96-well plates to expand clonal populations from individual cells. The instrument is in a bio-safety cabinet for safe clean sorting of human cells. You can achieve a very high purity (>95%).
This technology is optimal for separation of any sample in single cell suspension (peripheral blood, PBMCs, disaggregated tissue, cell lines). For most downstream assays, the cells will need to be sorted fresh (live) on the day of sample collection.
Magnetic Bead Cell Enrichment
Cells can be enriched from heterogeneous populations using antibodies coupled to magnetic beads with the Miltenyi AutoMACS system. This system allows both positive selection and negative “untouched” selection of cell populations. Compared to FACs sorting, with magnetic enrichment you can typically start with a smaller cell number and achieve a higher cell yield in a shorter amount of time. Depending on the separation kit and markers used, this technique enriches for populations with typical purities near 90%. Please see the Miltenyi website for available reagent kits.