Sulforhodamine B (SRB) colorimetric assay for cytotoxicity screening

Sulforhodamine B colorimetric assay for cytotoxicity screening. I did not add any voice descriptions in this video. You can find more information on this assay here:
www.ncbi.nlm.ni...

[@pedroalarconzapata9534]

Thank you for your work!

[@j.n4913]

May I ask another question?
If I want to have a baseline, that is I want to have a OD at day 0, how to do this?

[@memo4hope]

Thank you so much it was so helpful.. I just was wondering what will different in suspension cell lines??

[@8io6eek]

+Rose mary Thank you for your comment! I haven't tried with suspension cells lines, but my colleague did. Instead of aspirating the medium out of the wells after treatment, add 50 uL of 80% TCA to each well and refrigerate for 4 hours. The cells should be fixed to the bottom of the plate and proceed as usual. Good luck!

[@memo4hope]

Thank you so much.. I will try that next week and I'll tell you how it goes. :-D

[@luveni13]

Thank you for the video, it was indeed very helpful. What solvent did you use to dissolve trichloroacetic acid?

[@kamelsaleh8728]

Thank you.. excellent effort

[@konstans415]

Hello!Thank you for the video.After adding the Tris Base I noticed that my cells stayed intact and in the bottom of the wells.Is that normal?

[@8io6eek]

Yes, that is normal. The TCA fixes the cells to the bottom of the plate.

[@konstans415]

@@8io6eek thank you!

[@olefaulukitchen9376]

well illustrative, thank you

[@cigdemozen5005]

Hi,
Thank you for the video, it is very helpfull. I wonder, why do you sonicate the drug solution before you use?

[@8io6eek]

+Cigdem Özen Hi. My drug was a powder. Sonication can help make the drug more soluble. I wanted to make sure my drug is completely dissolved so I sonicate it.

[@cigdemozen5005]

Thank you

[@chamalikodagoda7680]

Thank you so much

[@Digitaleducation711]

How TCA fixes the cell in the bottom . what does it do to cells

[@8io6eek]

With 10% TCA, it fixes the cell monolayer so the SRB dye can bind the protein components of the fixed tissue.

[@olefaulukitchen9376]

But which cellular component does SRB binds to?

[@8io6eek]

proteins

[@jinx00471]

Excellent

[@j.n4913]

Can we use tca to fix cells for days, and at the last day to wash up tca and dye all plates together?

[@8io6eek]

Hi, my colleague left the TCA for 1 week and the data was still fine. Good luck!

[@drmostafamahmoud4620]

Thanks alot for your work, How long time SRB assay takes? Regards.

[@j.n4913]

@@drmostafamahmoud4620 it depends on your demands. You can even make a 146 hours experiment.

[@drmostafamahmoud4620]

Greatly appreciate your help. Thanks.

[@8io6eek]

@@drmostafamahmoud4620 It depends how long you want to treat your cells. Usually, the 1st day you will seed your cells. The last part to fix, dye and wash the cells can take up to 1.5 days.

[@AnhNguyen-jl6ul]

Thank you for your video

[@8io6eek]

You’re welcome! Hope it’s helpful!

[@eduardomoraisberetta7909]

muito bom!

[@user-nq2qp8fg1k]

Is it okay to wash with tap water after acetic acid washing?

[@8io6eek]

김한솔 yes

[@user-nq2qp8fg1k]

Can you guess why I got minus values of 'cells killed'(=100-100*meanODsample/meanODday0) T_T...?

[@8io6eek]

I usually do the following: (treated cell OD/control non treated cell OD)*100 = % cell viability

[@xiaolongwang6657]

so nice

[@8io6eek]

Thank you!