Hi Monalisa,

I did flow cytometry using the same probe (DCFH-DA). I had not problems with Ehrlich cells auto-fluorescence. Maybe, you should check the lasers parameters.

If you need something, send a message.

Good luck

Hi Marcelo!

Thank you very much by the answer.

By the way, do you still have the protocol?

I´m still testing the adequate amounts, guiding myself by the INVITROGEN protocol...

I´d like some "light", if possible!!

Thank you again.

Hi Monalisa,

find attached the protocol used by me.

Marcelo

Sorry, I forgot to attach.

Hi Marcelo!

I see you fix the cells to do the equipment´s adjusts.

I wonder if it was my mistake...

I´ll do the experiments and tell you what happened!

Thank you!!! See you later!

Hi Monalisa

You can check following reference also.

Mosiman, V. L., Patterson, B. K., Canterero, L. & Goolsby, C. L. Reducing cellular autofluorescence in flow cytometry: An in situ method. Communications in Clinical Cytometry 30, 151-156,

-vish

Selection of buffer is also important. You may avoid PBS or Tris buffer and use MES buffer with sorbitol.