Common sorting rules:
The nozzle size should be at least 5 times greater than cell diameter
Total event rate (including debris and dead cells) should be no more than 1/4 of drop frequency
Our default settings for all cell sorts using AriaII are: 100 µm nozzle, 20 psi pressure and 30 kHz drop frequency. You can typically expect ≥80% sorting efficiency and ≥90% purity when running ≤5 million total cells per hour using purity mask setting.
Non-homogenous samples, debris and faster flow rate often decrease the sorting efficiency and the purity of the recovered populations.
Observed cell recovery (manual count under light microscope) may be significantly lower than instrument reported sort efficiency if e.g. drop delay is poorly calibrated, cells die during sorting process (pH and pressure changes, drying on tube wall) or cells are lost in downstream processing steps(tube and buffer changes)
Recovery volume is 3 mL (of PBS) per 1 million cells
On special request we can speed up the sorting process using 70 µm nozzle (mostly for lymphocytes) or improve side stream quality using 130 µm nozzle (for unusually large cells).