ETHIDIUM BROMIDE STAINING OF TRYPANOSOMES

(This is a rudimentary protocol that I have used. It stains the nucleus very nicely, but kinetoplast staining is not that great, and fades quickly. I am sure their is plenty of room for improvement.)

1) Stain 10 min with 2 mg/ml Ethidium Bromide in PBS. (Yes, 2 mg/ml final!)

2) Wash 2x 100 ul PBS.

3) Resuspend in PBS. (Volume depends on # cells. Usually, 20 - 50 ul is good.)

4) Let sit at RT for 5 - 20 min, then examine. 20 min is good, but can go for 1 hr as needed.

NOTES:

Be sure to start with a good size cell pellet, because you tend to loose some at each wash step.

Pellet cells in u-fuge 3-4 min @ 8000 rpm.