How to Use the Microplate Reader

By Megan Forman, Luke Serensits, and Melissa Mcgroarty

• Turn on the power box (small rectangular box next to the plate reader).

• Turn on the Plate reader.• If the PC is not turned on,

turn it on and log into Bio421 (password is protein).

Turning on the Plate Reader

• After the plate reader and computer are turned on open Gen 5 1.10.

• Click on Protocol ( under Experiment).

• Once the protocol is opened click on procedure.

Starting Gen 5 1.10

• After double clicking on procedure, click on the plate type arrow and choose what plate you are using.

• Dr. Li will tell you what type of plate he is giving you for your experiment

Define your plate

• After picking what type of plate you are using double click on read.

• Select the areas where your samples are located. Make sure your plate reads continuously. The platereader can not read disconnected sections of the plate.

Read

• Set detection method to fluorescence.

• Read type is set to endpoint or spectrum (dependent on experiment).

• Read speed should be set to normal.

• Set the excitation and emission wavelengths and select the 9.0 nm band width (Dr. Li will give guidance on how to set them).

• When doing a spectrum select emission/ excitation step of 1 nm.

• If the samples show very high fluorescence, the sensitivity must be lowered.

• Lastly set the top probe vertical offset to 8.00 nm and click ok.

What Kind of Experiment?

• Shake needs to be set to 3 seconds.

• Click on shake so it is highlighted and drag the shake in front of your experiment (so that the plate is shaken before the machine reads the plate.

• Click on validate to make sure the sequence is ok and proceed by clicking ok.

Shake the Plate

• Click on the plate layout icon to tell the machine where the blanks and samples are located.

• Blanks and samples must be in the area selected in the full plate icon.

• After this is completed click ok.

Plate Layout

• The protocol that you created needs to be saved on the expansion I: drive (click file to save the protocol).

• Remember, make sure to only save the protocol on the expansion I: drive!

Save your Protocol!

• After you saved your protocol click on the new experiment icon.

• Next, click on the protocol that was made.

• Click on the read plate icon and press read.

• After read is pressed save your results to the expansion I: drive and click save.

New Experiment

• After saving your plate a Load Plate window appears.

• DO NOT CLICK OK RIGHT AWAY!

• First, load your plate correctly (A should be in the top left corner) and then press ok.

Loading your plate

•After the plate reader is done reading your samples and blanks your data can be analyzed.

•Click on the excel icon to export your data to excel.

•After the Data is obtained it can be manipulated to graph different things such as Intensity vs. Concentration.

Obtaining Data

•After results are obtained, take the plate out of the plate reader.•Make sure to close the plate reader door by pressing the small button above the power switch.•Next turn of the plate reader and then shut off the power box (small rectangular box). •Also, if needed turn off the computer.

Clean Up