I really appreciated this video - thank you for posting. We don't consider the centrifuge to be a 'clean' location, for this reason we wipe down the buckets before bringing into hood every time :)
Thank you very much for this very nice video. I am currently following the same process but with sheep blood. The point is I don't have 4 layers after the first centrifuge. Does anybody knows if the type of blood plays an important part in this experience ? thank you to reply me
Great video. I am wondering if there is 10ml blood, can more PBMCs be isolated using 5ml blood plus 3ml ficoll once for two times or total 10ml blood plus 6ml ficoll for one time?
You see, even after decanting the supernatant, the cell pellet remains immersed in around 200 microliters of media. Flicking the tube resuspends this pellet in the remaining media and then you proceed to top up this cell resuspension with 1 mL of basic media. You then count your PBMCs and correct for whatever concentration you need for further experiments or storage.
I would very much like to have the video translated into french with captions. This is something you can aid with. Unfortunately, it is outside of our current possibilities.
not bad but one should NEVER use that kind pipet to recover the cells from the mononuclear layer. I always teach people to use a 1 ml disposable 12" pipet for this! Or, use elongated 10" glass pasteur pipets with a bulb and cotton plug. sterilized of course. NEVER use that P-1000 you used because it is way too short and you are in danger of contaminating the tube, and it is just NOT long enough! :)!
wonderful protocol, just what I really need. Thank you very much
I really appreciated this video - thank you for posting. We don't consider the centrifuge to be a 'clean' location, for this reason we wipe down the buckets before bringing into hood every time :)
Thank you very much. This is very helpful.
Thank you so much this help me a lot
So helpful 😃
They seem to be using extra long pipette tips, probably to avoid this and none of the actual pipette is that near the tube. You have a point though.
very nice video , thanks a lot .
Thank you very much for this very nice video. I am currently following the same process but with sheep blood. The point is I don't have 4 layers after the first centrifuge. Does anybody knows if the type of blood plays an important part in this experience ? thank you to reply me
I've always recovered the PBMCs using sterile disposable pasteur pipettes (plastic).
Yes, we have done so too and works just the same.
@@TanoMac73 usted tiene el protocolo completo? Podría contactarme por favor? doctorraulortiz@gmail.com gracias de antemano!
To good for words ❤️
Great video. I am wondering if there is 10ml blood, can more PBMCs be isolated using 5ml blood plus 3ml ficoll once for two times or total 10ml blood plus 6ml ficoll for one time?
Processing volumes depend and are limited by the capacity of your centrifuge buckets and adaptors.
Without cooling ?
I cannot understand the last step about resuspend pbmc's..can anyone help?
You see, even after decanting the supernatant, the cell pellet remains immersed in around 200 microliters of media. Flicking the tube resuspends this pellet in the remaining media and then you proceed to top up this cell resuspension with 1 mL of basic media. You then count your PBMCs and correct for whatever concentration you need for further experiments or storage.
ممكن حد يحط المراحل كلها بالفرنسي
I would very much like to have the video translated into french with captions. This is something you can aid with. Unfortunately, it is outside of our current possibilities.
not bad but one should NEVER use that kind pipet to recover the cells from the mononuclear layer. I always teach people to use a 1 ml disposable 12" pipet for this! Or, use elongated 10" glass pasteur pipets with a bulb and cotton plug. sterilized of course. NEVER use that P-1000 you used because it is way too short and you are in danger of contaminating the tube, and it is just NOT long enough! :)!
That is exactly why we use extra long pipette tips
I think it's quite clear that you CAN do it lol she just did it