This video will show you how to use the SpectraMax M2 plate reader
To turn on the machine, switch the power button located on the back of the machine to the on position.
Then double click to open the SoftMax Pro-6 program on the computer.
a pop-up screen will automatically appear
First select the “choose different instrument” icon. Make sure to refresh until the name of the machine appears. Then click “Ok”
Next, select the “configure your acquisition settings” icon.
There are several different options with “Endpoint” and “Kinetic” being the most frequently used.
The endpoint option is to be used if you only need a single reading, while the Kinetic option allows multiple readings with customizable reading intervals.
You can also measure fluorescence levels in this plate reader
When analyzing fluorescence samples remember to use black opaque 96 microwell plates.
For demonstration purposes, here we will only show how to configure the Kinetic read option.
First, click on the kinetic icon. A list of all the adjustable parameters will appear in the left panel.
Then select the number of wavelengths you would like to read your sample at.
Up to 6 can be used, but here we will use 1.
Next select the wavelength you would like to use
600 Nanometers is commonly used to measure bacterial cell concentrations
For plate type select the first option which is the 96-well standard clear bottom plate
For plate area drag and select the wells you would like to read
For timing the total runtime and reading intervals can be set, the timing here is shown in hours minutes seconds
After setting your timing parameter the total number of readings will appear in the bottom
There is also an option to shake your plate prior to each reading
This is good to include as it helps disperse any cells that may have settled to the bottom of the well.
to add the Shake option select both boxes
5 & 3 seconds for before and inbetween reads are the default settings.
You can increase or decrease these periods as needed for your experiment.
“Speed mode” provides faster well reading but can result in less accurate results.
It should be used when shorter read intervals are applied
In “more settings”, it is very important to unclick the “calibrate” box, or condensation will form on your plate lid leading to inaccurate absorbance measurements.
After you have configured your acquisition settings, select “configure plate layout”.
This program allows you to pre-select where your standards and or samples are located.
To do this, drag and select the area you would like to edit.
Then click add and enter a name for the selected area.
Make sure to also indicate whether the selected area is for standards or unknowns
next select plate blank or group blank
If you are doing a single analysis you will likely only need one blank for the whole plate, in that case select the well and click on plate blank.
however, if you are performing more than one type of test on a single plate, you will need to create groups that each have their own blank.
To create a group blank, simply select a designated well for that group and click on the group blank option.
The series option allows the user to input specific concentration values if a standard curve is used. In this example, the standard curve runs from left to right so that option is selected.
The starting value of our standard curve is 100000 and it reduces by 10 folds every step.
The temperature of the plate reader is also adjustable
To set the temperature click on the temperature icon located in the top panel
Select on then proceed to set your desired temperature
Click ok when finished
Please note the temperature can only be set at ambient temperature or above, it cannot cool below room temperature
Make sure to monitor the temperature and to not start your experiment until the desired temperature has been reached
to input plates first click on the open flash close icon in the top panel
the instrument door will automatically open
Then carefully load your plate into the machine and press the same icon once again to close the door
When you're ready select the read icon the machine will start running
To view the progress of your readings click and drag to select the wells you want to view
Then hold down the cTRL key while double clicking the selected area to bring up the progress graph
At anytime you wish to stop the reading click on stop
When the readings are complete click on the play icon at the top left corner to export your data
check the box for Plate 1
You can select raw to export the unedited version of your data
or you can select reduced to export your data with the blanks already subtracted
Make sure to select columns output format for Kinetic run and Click ok when finished
be sure to change your file type to excel files before clicking save
To shut down the machine ensure the plate door is closed before turning the machine off using the switch at the back
Thank you for watching