We are experiencing different problems pelleting fibroblast:
a) From the resuspension of the comercial vials (after defrosting, we resuspended in 5ml complete DMEM + L-Glutamin + FBS + Streptomycin + Penicilin)
b) From a suspension of a 25mm2 flask at confluence (same medium).
In both cases we used 50ml falcon tubes and 25ºC.
- We don´t see any pellet by centrifuging at 1500rpm/254 rfc
- We see just a very tiny deposit when centrifuging at 1900/400 rfc. But we are afraid the cells could suffer.
Any recomendations? Thank you very much for all your help.
Dear Dr. Ana Lopez,
I feel the selection of an appropriate centrifuge tube size may solve this problem. Depending on the volume of your sample, you may choose the most appropriate tube size to ensure precise and effective separation as it is a critical factor in optimizing the centrifugation process.
Using a tube that is too large for the sample volume can lead to inefficient separation, as the larger surface area may cause the sample to spread thinly, reducing the sedimentation force on each particle.
Therefore, matching the tube capacity closely with your sample volume is essential for achieving optimal separation efficiency.
Have you tried using 15ml centrifuge tube instead of 50ml tube for a small volume of 5ml? I feel it should work at the same speed of 1500rpm.
Regards,
Malcolm Nobre
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Biology
- Biomolecules
- Nucleic Acids
- RNA