Controls: these tubes do not need to have as many cells as your real sample, you can do as low
as 500k cells per tube for controls

Negative (unstained) or parental cells (NO FLUORESCENT PROTEINS)
Single Color Controls (one color/fluor per tube)
Viability Control – same as the negative or parental cells - we will add the viability dye in the core once you are here

For sorting, since cells are NOT FIXED, we add viability controls in the core – we have DAPI (violet), 7AAD (yellow green) and PI (yellow green) – these are all DNA dyes so if your cell is alive and the membrane is intact then your cells will be NEGATIVE and the dye will NOT incorporate into your cells
We do not require people to use viability dyes but it is strongly encouraged - dead cells tend to have higher autofluorescence, leading to false positive signals so excluding dead cells allows for better resolution of your population

Resuspension buffer (what you will bring your sample in, with the recommended cell
concentration as above)

1X PBS + 2% serum/BSA – please add 2% serum, this significantly helps keeping the cells
more viable
Bring 10-30 mL depending on how many samples you have

Collection media (what we will collect your sorted sample into)

Media WITH A MINIMUM OF 20% SERUM/BSA – the cells that are sorted are in PBS with low serum and will dilute your media as more cells are added
Put your media into your own collection tubes and bring them already made to the core

If you are sorting into a 15 mL conical tube (most common) then add 3-5 mL of collection media per tube
If you are sorting into a 96 well plate then add 100 uL of your media – then once back to the lab add another 100 uL of fresh media on top