Initial OD Calibration

Experiments
1

Hi!

I am planning out my first OD calibration. I plan to use cells directly, and wanted to confirm a few other pieces of my workflow. Below are my questions:

  1. It looks like the current eVOLVER code will calibrate both photodiodes at once, correct?
  2. I plan to start with the lowest concentration, and pipette in more cells to the same vial for increasing OD points. As long as the vial has at least 25 mL, it should be ok if the volume increases a little with each point, since I will also be measuring the OD independently, correct? This way I don’t have to change the vials as much.
  3. I plan to use a Quick Drop reader with a cuvette for the independent OD readings. What do folks use? Is there something more efficient I should consider?
  4. Lastly, how often do you recommend that we calibrate temperature and OD? Before every experiment?
  5. How low can/should I go on the low OD side?

Any other recommendations?

Thank you for the help!
Katie

2

Here is a dated post (using old MATLAB scripts) on what should be done. Generally, the calibration method is similar. I hope to update w/ another post soon.