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Sample guidelines for the cell sort
1. Human samples must be serology tested (HIV and Hepatitis B negative) before brought to the FCRC.
2. Samples suppose to be single cell suspension (SCS) only (i.e. no clumps).
- To prevent clumping during the sort, filter the SCS through the mesh (we suggest to use the 5ml Polystyrene Round-Bottom Tube with Cell-Strainer Cap, Falcon #352235);
- You could also add 0.5 mM EDTA to your buffer solution;
- Use 1-5% serum for the buffer.
3. All the samples to be used at the FCRC should be placed ONLY in the following types of tubes:
- 12 x 75 mm 5ml Polystyrene Round-Bottom Tube non-sterile, Falcon #352008;
- 12 x 75 mm 5ml Polystyrene Round-Bottom Tube with Cell-Strainer Cap, Falcon #352235;
- 12 x 75 mm 5ml Polystyrene Round-Bottom sterile Tube with the Cap, Falcon #352054.
4. Please provide the controls - unstained and single color controls. Preferable concentration of the controls is around 1 x 106cells/ml, volume 0.5-1.0 ml.
5. The sample concentration for the sort should typically be around 10-15 x 106cells/ml. If you have fewer then 5 x 106 cells put them into the minimal volume of 0.5 ml.
6. Please provide extra 15 ml of the buffer you use for the cell samples.
7. Please provide collection tubes for the purified (post-sort) cells.
- For the post-sort tubes use eppendorf tubes or any type of 12 x 75 mm tubes (made from glass, polypropylene, polystyrene, etc.);
- In order to prevent cells sticking to the sides of the tubes, pre-coat the tubes, filling them with the serum for 30 minutes before the sort;
- For the sterile sort the post-sort tubes should be also sterile.
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