Sorting Conditions

Our facility offers four different sorting conditions to accommodate various cell types:

📌 70 µm / 70 PSI – For splenocytes, thymocytes, PBMC, whole blood, and bone marrow cells

📌 85 µm / 45 PSI – For transfected B or T cell lines, primary lymphocytes, and bone marrow cells

📌 100 µm / 20 PSI – For fibroblasts, primary cultured cells, endothelial cells, and adherent cell lines

📌 130 µm / 16 PSI – For dissociated tumor cells, large cell lines, and fragile cells

📌 Basic Sorting Buffer

Prepare your samples in 1X PBS or HBSS (without Ca²⁺/Mg²⁺) with 1% FBS and 1 mM EDTA.

🔹 Modifications for Specific Cell Types:

✔️ Sticky Cells – Increase EDTA to 5 mM to reduce clumping.

✔️ Sensitive Cells – Add 25 mM HEPES to stabilize pH.

✔️ Samples with High Cell Death – Add DNAse to prevent aggregation.

Sample Collection

Cells can be collected in various types of tubes or plates:

• Eppendorf tubes
• 5 ml FACS tubes (ideally polypropylene as cells adhere less than in polystyrene)
• 15 ml conical tubes (2-way sort only)
• Multi-well plates (6, 12, 24, 48, 96, 384) (1-way sort only)
• Slides (1-way sort only)

Coat the recovering tube/plate with media or PBS before arriving. You can add roughly 10-20% of the receptacle volume (e.g. 2ml in a 15ml conical tube). Consider adding a higher concentration of FBS to the recovery media for sensitive samples.

The collection tubes can also be kept at 4°C during the sort. You can decide the collection temperature on the day of your sort and tell the operator.