I have isolated human neutrophils (from blood) in the inactivated condition for parasite infection study. I need to fix the cells for staining protocol. I have used 1% and 4% paraformaldehyde. I am getting more ghost cells during staining protocol. What should I use for fixing cells? Also I need it to grow on coverslip. What is the method used for adhering neutrophils on glass coverslip?
Hi,
Even I'm observing a lot of Ghost cells during staining. Did you find a way out? What conditions did you use to fix your cells?
Thanks,
Hi Riva,
If you want to fix PMNs for ICC then fixed cells with 4 or 3.7% PFA at 4 degree Celsius for 30 minutes. Used zero acceleration and break for centrifugation when cells are live. Give minimum jerk to cells during resuspending pellet. Avoid using pipette for resuspending cells. Whenever you fix cells for microscopy then try to use the cells within 3-4 days after fixation.
This procedure works well with me. Try same.
Best luck!
Durgesh Pitale.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line