Dear Ziad Salem!

Please you look at this chapter:

Fish cell lines as a tool in aquatic toxicology

https://link.springer.com/chapter/10.1007%2F978-3-0348-8853-0_1 (attached)

Figure 2. Optical density measurements with crystal violet (CV), neutral red (NR) and MTT formazan (MTT) stain as a function of cell number. Absorption was read in a microtiterplate reader at 590 nm for Cv, and at 540 nm for NR and MTT (adapted from Lenz et aI., 1993). (page 6)

Hi Ziad Salem Fouda

You are correct in thinking that you need to perform a statistical test. For this, a simple student's t-test would suffice. However, I notice you do not appear to have a control (cell viability at 0 M copper).

Cell viability is essential for ensuring that your added compound does not interfere too much in subsequent experiments. Now that you have this data, you can run further experiments on your cell lines to determine other properties such as proliferation or adherence.

Best Wishes

Dear Zaid,

It is very nice pilot study.

Is it sour results or after calculation since you have to delete absorbance of reference wavelength (e.g. 630 nm) from absorbance of test wavelength

(e.g. 570 nm). Base on that results you can prepare a graph. In my opinion, you should add more concentration between 0.1 and 1.0 M e.g. 0.25, 0.5, and 0.75 M and than calculate IC50. Since now you have only one result below 50%, if you add more concentration the IC50 calculation will be more precise. Please also remember that if you will add more concentration, than please do not use t-test but one-way ANOVA.

Best wishes

Sorry i forgot to add in my control value, The average absorbance reading was 0.49356 (100% cell viability).

Dear Alexandr Chernov

Thank you for the paper I shall give it a read later today

Dear Jack Hopkins

For a students t-test, how is it that I compare more than 2 sets of data? (Would I include the control in my calculations?)

Dear Michał Otręba

If i were to do an ANOVA, would I include my control result too in the calculations?

Sorry for the inconveniance.

Dear Ziad Salem Fouda

Compare between your control and a particular concentration (only two datasets). However, if you want to compare against all samples simultaneously, Michał Otręba is quite right and you should perform ANOVA.

Dear Zaid,

Yes if you will use ANOVA you have to include your control results. It is the best way to calculate absorbance into % of control. To notice differences in control and see error bars on graph in control samples you can do it like that (for control):

Mean absorbance is 100%, and than recalculate all of the control samples results into %

I hope it will help you :)

Dear Zaid,

Please see this video

Calculation of LC50 in Cytotoxicity Experiment through Excel in Hindi - YouTube

www.youtube.com/watch?v=LeSqqPENQ78