This was great 👍🏻

Nice explanation. Thank you

Awesome 👌

thanks so much

really helped in my campbell biology exercise

Thank you sir very helpful

Can you please tell me how much PI and Rnase should be there? Should we add sodium citrate with that?

Hi I want to measure cell counting of sulfate reducing bacteria. Can you share any suitable protocol ?

thankyou so much

Thanks

Thank you so much. Would Dapi staining work the same?

@thatdudenate22

3 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.

@thatdudenate22

3 жыл бұрын

but just to be clear you are still able to use DAPI but that phase might be a little inaccurate

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

How can we count the cell?? In cell cycle

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.

@michaeldgn9813

8 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

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.

How many cells is required for an analysis?

@natureabioros8686

Жыл бұрын

Several thousand at least for Flow Cytometry.

@elisabethpepsiparty5735

6 ай бұрын

10k and it will take a LONG time with this little cells

@ 0:57 I swear I thought that was my phone😂

Why are we degrading RNA during cell cycle analysis?