I have cultured human cancer cells with carbon nano particle. After culture, I wanted to collect the cells to do FACS and some others. However, the nanoparticles affected the result. How could I separate the nano particles from the culture medium?
The particle is about 0.5 um.
Thank You very much, Mohammad Kamal!
Did it work well for your samples? I will try with your suggestion. But, with that speed, does it make the particle conjugated to the cells?
You could remove the media first, wash the cwlls a few times with pbs, and then detach them for facs analysis in fresh media
The best solution would be doing what Dr. Kamal and Dr. Chou suggested, in this order and 2 to 3 times. (Something called repeated washing :)
Are your cells adherent or grown in suspension? If they are adherent, then Dr. Chou's suggestion would be the best route. If the cells are grown as a suspension culture, then a low speed spin (500-800 rpm for 3-5 min) would sediment the cells leaving the majority of small nano particles in the supernatant. Repeat this 1-2 times before resuspending in fresh media for FACS analysis
Try density gradient centrifiguation, i.e. at low rpm u can have cells seperated first leaving free carbon nanoparticles in the medium....
Many thanks to Lea Chou, Tina Leisner and everyone! the cells are adherent.
Please try to go for centrifugation at low rpm because at the high rpm that is above 1500 rpm nanoparticles will start forming a pellet.
Thanks Arpita Gupta!
I tried with 500 rpm for 30 seconds since the nanaparticle is easily down. It is pretty good for the FACS analysis.
Sincerely,
Dear Quyen Nguyen,
I'm happy to know that it worked... :)
I wanted to know that what was the objective behind culturing the carbon nanoparticles with human cancer cells?
Best of Luck for the further studies.....
It is better to apply magnetic nano-particles e.g.:
Article Magnetic Gold Nanoparticles in SERS-Based Sandwich Immunoass...
To immobilize some antibodies
Article Determination of antibodies against human growth hormone usi...
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