How to graph or plot MTT assay data - (Jan/01/2013 )

this is example of data collected from elisa plate reader. i dont know hoe to plot the graf of % cell viability vs log concentration

I will say what I would do if I were u
for % of viability I calculate it as abs of test divide by abs of control (no treatment) X100
(get average of Abs for test repeats in ur exp / by average abs of repeats of control)
Each experiment should be repeated 2-3 times at different days or more, so I would calculate STD from my these repeats ( if I do the experiment 3 times) I would calculate % of vaibility for each time as previously mentioned then calculate standard deviation or Standard error for these 3 times,
the i guess X axis would be log conc, and Y axis would be % of viability..
This I would do if I were u

Hello,

I would also appreciate some help with this. I do MTS assays to measure the activity of certain compounds. I do each trial with 4 replicates for each group and then normalize the abs of each well with the average abs of the control. After that I calculate the average and standard deviation for each group. I then repeated the experiment 2 more times for a total of three trials, but now I end up with three averages (each having a standard deviation) and I can't figure out the proper way to calculate the final average and standard deviation. I have tried various search terms in google and could not find anything that match exactly with what I need.

For example, the data that I got for drug A at a certain concentration is (data are % of control): Trial 1: mean = 90 , SD= 5; Trial 2: mean = 94, SD = 4; Trial 3: mean = 89, SD = 3. How do I calculate the overall mean and standard deviation of these values?

I was told that I can just propagate the errors of each trial. Is this OK to do?

It would be a lot easier if I could just average the abs for each treatment group, normalize to the control and calculate the average and SD of the three trials, but what happens to the error associated with the abs of the replicated wells?

Would greatly appreciate a response if anyone has any experience with this type of calculation.

Thanks!

Ms VV on Wed Mar 6 01:47:17 2013 said:

Hi DRT, thank you for responding! Just to clarify, your suggestion is to enter the % viability of each well for each trial and do an ANOVA to test for statistical significant differences? So I should enter the data as follows?

Trial 1 Trial 2 Trial 3
% well 1 % well 1 % well 1
% well 2 % well 2 % well 2
% well 3 % well 3 % well 3
% well 4 % well 4 % well 4

If there are no significant differences, then I can just use all the values to calculate one average and standard deviation?

That seems correct, most software will want the results in a single column with a tag denoting which trial in an adjacent column.

good luck

Ms VV on Wed Mar 6 01:47:17 2013 said:

Thanks DRT! I'll try that and see.

Hi Peniel! Yes, I use 96 well plates. I usually incubate for 48 h without replacing the medium and have no noticeable decrease in volume by looking at the wells. There is a decrease of 0.1-0.2 mL when measured. I incubated a few times for 72 h and also did not noticed a big decrease in volume. I use 0.1 mL per well. The wells could hold up to 0.3 mL, maybe you could use a higher volume if you are not already doing so. Sorry, I don't really have any suggestions other since I haven't had that issue.

Peniel on Fri Mar 8 11:01:42 2013 said:

DRT on Sun Mar 10 18:57:10 2013 said: