How many cells per well if we use a 96 plate? 72h treatment... I know it varies, but my cells grow a lot, I wonder what you would suggest haha
@chaithu9132 жыл бұрын
Thank you sir very helpful
@sambitpradhan6292 Жыл бұрын
Can you please tell me how much PI and Rnase should be there? Should we add sodium citrate with that?
@hanumakumar8997 Жыл бұрын
Why are we degrading RNA during cell cycle analysis?
@AnimeshGoel10 ай бұрын
How can we count the cell?? In cell cycle
@vihaancubejunghare56145 ай бұрын
really helped in my campbell biology exercise
@fidahussainshigri56662 жыл бұрын
Hi I want to measure cell counting of sulfate reducing bacteria. Can you share any suitable protocol ?
@PRIYANKAGONEMAD Жыл бұрын
Why doesn’t S phase have a higher peak than G0/G1? Cells synthesisze new DNA in S phase so there much be more DNA that PI binds to.
@michaeldgn981310 ай бұрын
The peaks on the y-axis are the amount of cells, so the high peak at G0/1 is showing that there are lots of cells at this stage in the fell cycle. Along the x-axis is the DNA content so S phase is showing more DNA content than G0/G1, and G2 is showing more DNA content than S and G0/G1
@charlesidu-okojokwu97325 ай бұрын
Awesome 👌
@jono43273 жыл бұрын
@ 0:57 I swear I thought that was my phone😂
@fallseason51632 жыл бұрын
This was great 👍🏻
@quintabrownderiona7023 жыл бұрын
Thanks
@mamostamuhamad73263 жыл бұрын
thanks so much
@zahrarashno45863 жыл бұрын
thankyou so much
@tomwinkler2943 жыл бұрын
Thank you so much. Would Dapi staining work the same?
@thatdudenate223 жыл бұрын
DAPI stains the nucleus but doesn't work well on apoptotic cells. Propidium Iodide (PI) works better if you are trying to analyze all cell cycle phases.
@thatdudenate223 жыл бұрын
but just to be clear you are still able to use DAPI but that phase might be a little inaccurate
@IScreenshotNFTsАй бұрын
I used DAPI. And it works pretty well, but at very high concentrations, of 20ug/mL. Internet recommends 0.1-1ug/mL, but thats wrong. I strongly suggest to use PI, works better.
@ridongfeng89242 жыл бұрын
How many cells is required for an analysis?
@natureabioros8686 Жыл бұрын
Several thousand at least for Flow Cytometry.
@elisabethpepsiparty57358 ай бұрын
10k and it will take a LONG time with this little cells
@tomwinkler2943 жыл бұрын
You resuspended in 2 ml ETOH?
@natureabioros8686 Жыл бұрын
For a fixing protocol where the cells are killed but preserved for imaging. This way you dont have to worry as much about decay of the cells before your analysis. Similar in principle to paraformaldehyde (PFA) treatment of tissue sections or microscope slides.
@natureabioros8686 Жыл бұрын
Presumably you dont have to though if you want to keep your cells alive. Prob put them in serum free media till you need to analyze, then prob swap out your media with PBS for less background signal.
@natureabioros8686 Жыл бұрын
This is also necessary for Propidium Iodide (PI) staining since it is membrane impermeable in live cells. Thermo has some nice non-toxic dyes that distribute throughout the cytoplasm and are membrane permeant to live cells. See CellTrace. They have three kinds, Violet, Yellow, and Deep Red.