Cell Preparation of Tissue Culture Cells.
Materials
- Trypsin or EDTA
- Flow cytometry staining buffer
- 15 or 50 mL conical centrifuge tubes
Experimental Procedure
1. For cells that grow in suspension, decant the cells into a conical centrifuge tube and perform a cell count and viability analysis.
Proceed to Step 4.
2. For adherent cells lines, detach your cells from the plate, using trypsin or EDTA (10 mM in PBS).
3. In a conical centrifuge tube place the cells and perform a cell count and viability analysis.
4. Centrifuge cells (4-5 minutes at 1000-2000 rpm) and resuspend in the flow cytometry staining buffer so that the final cell concentration is 1x107 cells/mL.
References: http://www.scbt.com/protocols/protocol_08.pdf
http://media.ebioscience.com/data/pdf/best-protocols/cell-preparation-for-flow-cytometry.pdf
1. For cells that grow in suspension, decant the cells into a conical centrifuge tube and perform a cell count and viability analysis.
Proceed to Step 4.
2. For adherent cells lines, detach your cells from the plate, using trypsin or EDTA (10 mM in PBS).
3. In a conical centrifuge tube place the cells and perform a cell count and viability analysis.
4. Centrifuge cells (4-5 minutes at 1000-2000 rpm) and resuspend in the flow cytometry staining buffer so that the final cell concentration is 1x107 cells/mL.
References: http://www.scbt.com/protocols/protocol_08.pdf
http://media.ebioscience.com/data/pdf/best-protocols/cell-preparation-for-flow-cytometry.pdf