Survival rate in MTT (View forum version)



saraarasus

Posted 17 March 2009 - 05:03 PM

Hi,
Has any one know how should I calculate the survival rate after getting the OD from MTT.

Many thanks,
Sara

WOW

Posted 17 March 2009 - 10:04 PM

According to my understanding MTT assay in most cases reflects "cell viability". This term was somehow different from cell survival or cell proliferation. Therefore once you do this assay in a short time (within 24hrs), it mostly reflect cell death and survival. If the time is more than 24hrs, you should consider proliferation effect. Since I am not clear your experiment design, I could not exactly answer your questions. May it help you and good luck.

saraarasus

Posted 18 March 2009 - 02:46 PM

Hi WOW,
Thanks for your reply. What I did was that, I treated my cells for 24 hours with my protein. Then I did the MTT. I remove the media , then add 90micro litre of RPMI without phenol red, then I added 10microlitre of 5mg/ml MTT in the same media and incubate it for 4 hrs at 37 deg. Then I added MTT solvent and mix them and read the OD whithin 1 houre.
I have two question:
1-Is it the correct procedure?
2- What should I use as the high control?

Thanks,
Sara

cotchy

Posted 19 March 2009 - 06:24 AM

Hi WOW,
Thanks for your reply. What I did was that, I treated my cells for 24 hours with my protein. Then I did the MTT. I remove the media , then add 90micro litre of RPMI without phenol red, then I added 10microlitre of 5mg/ml MTT in the same media and incubate it for 4 hrs at 37 deg. Then I added MTT solvent and mix them and read the OD whithin 1 houre.
I have two question:
1-Is it the correct procedure?
2- What should I use as the high control?

Thanks,
Sara


Sara your protocol looks fine.

A positivie control of DMSO can be used (1 % or 0.1% DMSO dissolved in your normal media is sufficent to kill cells)

DRT

Posted 19 March 2009 - 03:26 PM

A positivie control of DMSO can be used (1 % or 0.1% DMSO dissolved in your normal media is sufficent to kill cells)


Wouldn't you need a higher concentration than 1% DMSO? ^_^

bob1

Posted 19 March 2009 - 04:00 PM

0.01% DMSO is sufficient to kill most cell lines, it dissolves essential cell membrane components.

saraarasus

Posted 19 March 2009 - 06:42 PM

0.01% DMSO is sufficient to kill most cell lines, it dissolves essential cell membrane components.


Thanks for your helpful reply.

Cheers,
Sara