Assay #2 – Assessing Cell Viability at the

EC50 – LIVE/DEAD Assay using Crystal

Violet
TA: Samantha Ramirez
Announcements
Lab Follow Up
 Lab Goals:
• You will make a dilution series using your compound and you will treat your cells
with them for 24 Hours.
• You should try and guess your EC50 concentration from the known
concentrations list using Crystal Violet stain.
Your compound will be used at the following concentrations:
Column 1 - 66 μΜ
Column 2 - 33 μΜ
Column 3 - 16.5 μΜ
Column 4 - 8.25 μΜ
Column 5 - 4.125 μΜ
Column 6 - 2.0625 μΜ
Column 7 - 1.03125 μΜ
 What you should know:
(+) Control: (-) Control:
What Exerts the desired effect? What Exerts the opposite
effect?

Vehicle: EC50:
The Diluent the compound is Half Maximal Effective
administered in? Concentration.
 What you will do today:
You will be working in the hoods:
• Prepare your compound dilution
series.
• Treat your cells as presented on
the right.
• Treat with your vehicle positive
and negative controls as
presented on the right.
 Lab Follow Up:
24 hours after cell treatment with compound:
• A member of each group will need to come back to add crystal violet
stain and take pictures of the cells under light microscopes as below:
 Example of Results:

1) The EC50 here has been

qualitatively analyzed at a
concentration of 8.25 μM.
2) Positive and negative
controls observed correct
responses.
3) Vehicle Control did not effect
the cells which means diluent
had no impact on the cells.
4) Therefore capsaicin at 8.25
μM effectively triggers 50%
cell death in A-549 cells.
ANY QUESTION?