Updated: 2023-04-07
Revision: B
Note: There is no particular protocol for spiking. This article provides tips that may improve the accuracy of your spike-ins.
Depending on the lab, some prefer to pick the cells and spike (more accurate).
Others do serial dilution of the cell suspension then spike (less accurate).
Here we use a method that generally gives good precision of the number of cells spiked (in the 100 to 500 range).
- Make sure the suspension is of single cells and not aggregates. Gently triturate to disperse the cells if necessary.
- Determine the cell concentration (do a quick pre-count of a diluted cell sample).
- Make an appropriate dilution so that the concentration is about 5 cells per microliter (100 cells/20 microliters)
- Use the appropriate volume corresponding to the desired number of cells to be spiked
- After spiking, recount at least 4 times, each with the same volume that was used to spike
- Determine the count, standard deviation, etc...