Flow cytometry on erythrocytes - Need practical advice (Dec/12/2008 )

Hi,
I would like to detect a certain protein on mouse erythrocytes. How much do I have to dilute the blood before doing the labeling? Are additional manipulations with the blood necessary? Is the labeling procedure like a standard labeling protocol? How about the settings on the flow cytometer?
Thank you for all helpful hints!
Regards

There are various cellular stages of RBC development. Are you looking to stain nucleated erythrocytes or terminally differentiated (non-nucleated) erythrocytes?

If you are still interested in this question, I can ask a colleague who frequently uses flow cytometry to study erythropoiesis.

Hi,
We are interested in terminally differentiated erythrocytes. But making a distinction between nucleated and non-nucleated erythrocytes on flow cytometry is not really our goal. It would be nice to have some practical advice regarding dilution of blood, centrifugation speed, incubation times and concentrations for antibodies, solutions for agglutination, settings on the flow cytometer etc. Thank you in advance.
Regards,
Sofie