Tips For Spiking Cells Into Whole Blood

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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...