Preparation of Single Cell Suspensions for Mass Cytometry
Materials and Instruments
1. Experimental cell lines
2. Phosphate-buffered saline (PBS)
3. 0.25% Trypsin-EDTA
4. Centrifuge
5. Flow cytometer
Preparation Procedure
1. Cell Washing and Harvesting
Begin by washing the cells with PBS. Following this, treat the cells with 0.25% Trypsin-EDTA to facilitate cell detachment and harvesting.
2. Suspension and Centrifugation
Resuspend the detached cells in PBS. Transfer the suspension to a centrifuge tube, and centrifuge at 300g for 5 minutes to pellet the cells.
3. Resuspension and Filtration
After discarding the supernatant, resuspend the cell pellet in fresh PBS. Filter the cell suspension to remove any aggregates and large debris.
4. Cell Counting and Dilution
Use a cell counter or hemocytometer to determine the cell concentration. Adjust the cell suspension to the desired dilution according to experimental requirements.
5. Preparation for Mass Spectrometry Analysis
Prepare a single-cell suspension tailored for mass spectrometry analysis. This step may require modifications based on the specific protocols associated with the mass spectrometer in use.
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