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| STEP 7 | Here is the tube after centrifuging pellet. Don't worry that the pellet is red and some red cells cling to the conical part of the tube. We will clean it up. (figure 7) |  figure 7 (click on image to enlarge) | |||||||||||||||
| STEP 8 | Remove and discard the clear supernatant down close to the pellet, but not too close. Some red cells are still under the pellet and the white cells are on top of it. (figure 8) |  figure 8 (click on image to enlarge) | |||||||||||||||
| STEP 9 | To remove the red cells we will make the solution hypotonic so that they will swell and burst open. The white cells are more robust and will not swell and burst, i.e. lyse. Our 0.9% sodium chloride, distilled water, and 3.6% sodium chloride solutions are kept cold in the ice bath and arranged for easy access. (figure 9) |  figure 9 (click on image to enlarge) | |||||||||||||||
| STEP 10 | First add 0.8 ml of 0.9% sodium chloride. (figure10a) This is normal saline which is istonic and there will be no swelling in this solution. Try to get the pellet at least partly suspended by vortex at this point. (figure 10b) |
STEP 11 |
If the pellet does not suspend easily, you can cut off the plastic pipet top to widen the orifice (figure 11a), then pipet up and down (figure 11b), until it is is suspended (figure 11c)
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