Checklist for FACS Cell Analysis/Cell Sorting
Preparation of cells for FACS analysis or sorting:
- It is possible to sort human cells, if the S1/L2 requirements are fulfilled (e.g. tested negative for HBV, HCV, HIV).
- A specific request is required, if the material contains nontested human blood or nonfixed cells from human tissues.
Explanation: The basis of the BD FACS Aria-II sorter is a microfluidic system which potentially generates aerosols. While the sample fluids are exposed to high pressures during the sorting process, the aerosols can be generated which contain potential HBV, HCV or HIV from the sample. Our device is equipped with an aerosol management system and therefore, can be prepared for these applications.
- We don't provide sorting of microorganism or of cells which have been in contact with micororganisms; thereby, we are able to provide the sorting of samples under semi-sterile conditions.
- We only can provide sorted cells which are contained in the original sample; a potential loss of at least a quarter of the cells should be calculated.
- Please provide the sort samples as stained single cell suspension in a concentration of about 1x10^7 cells/ mL; ideally, the cells are suspended in a PBS-Buffer mit 0,5% BSA und 2mM EDTA; during the sorting process any buffer will be replaced by semi-sterile PBS, which the machines operates with.
- To exclude cell clumping, please filter the cell suspension through a mesh of 40 µM pore size (available at Falcon or Miltenyi Biotec; ideal are 5mL Round Bottom Polystyrene Tubes with integrated cell strainer - Falcon #352235)
- Please provide tubes filled with medium to collect the sorted cells; it is recommended to supplement the medium with antibiotics as the sorting process is semi-sterile.
- It is possible to sort in to 96-well plates, but this should be communicated when the sorting appointment is made.
- Please provide staining controls and samples to set up the machine.