Measuring Cell Density Over Time

CAUTION: ALWAYS WEAR GLOVES WHEN USING THE PLATE READER AS ETHIDIUM BROMIDE IS USED IN THIS ROOM.

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1. Open the top of the plate reader, and adjust the wires so they are set to read absorbance (use the guide in the machine).
   
   
2. Log on to the computer as lab admin.
   
   
3. Open the program Fluostar Optima.
   
   
4. Within the program, log on as lab user.
   
   
5. Click on test setup, then test protocol.
   
   
6. Click on the protocol called OD timecourse. Here you can adjust the number of cycles and the cycle time (time between each reading). You can also adjust the gain, which should be set to 750 for absorbance readings. The absorbance should be set to 595nm.
   
   
7. Click on the Concentration Volume Shaking tab. Here you can change the shaking settings so that the plate is shaken after each cycle, for the maximum time of 300s with a width of 5.
   
   
8. Click on the layout tab. Here you can change the settings to change the number of wells on the plate. The start value should be set to 1 and set to increase.
   
   
9. Click OK.
   
   
10. Click the reader configuration tab and ensure it is set to absorbance.
    
    
11. Click on measure, then temperature control. Here you can adjust the temperature to the correct temperature for the bacteria being used.
    
    
12. Click OK.
    
    
13. Click the open plate drawer button and insert your plate. Then click close plate draw.
    
    
14. Click measure, then click measure from this menu. Select your protocol. Here you can save the name of your reading.
    
    
15. Then click the start test run button.
    
    
16. By clicking the button with the graph icon you can monitor the progress.
    
    
17. Once measurements have finished being taken, click on the button that shows all readings the machine has taken.
    
    
18. Double click on your reading and copy and paste the results into a new excel file.
    
    
19. Save the file.