Tune parameters tab

This tab contains tools that will allow you to determine the best paramters for your dataset.

Start by clicking on Start adding points button on the top-right of the tab.

You can add both true spots but also spots that could be detected but are not valid (e.g., dimmer spots that should not be counted).

To switch between adding true or false positives toggle Clicking on true spots (middle-right of the tab in the “Spots properties” section). In the same section you can also change the apperance of the spots or clear the selected spots.

The first parameter that you can visually tune is the Resolution multiplier in y- and x- direction. To tune this, activate it by clicking on Activate on the right of the plus button, then hover on one spot and press up/down arrows on the keyboard to adjust the circle size to the spot.

The second parameter is Spot minimum z-size (μm). To tune this, activate it by clicking on Activate on the right of the plus button. The viewed plane will switch to the zy plane to allow you to see the spots along the z-axis. As before, hover on one spot and press up/down arrows on the keyboard to adjust the length of the line to the height of the spot.

Once you add some spots, you can view their features in the “Numerical values of the last spot hovered with mouse cursor”. Select which features you want to view and hover with the mouse onto a spot.

This step is very useful to get a sense of what could be a minimum and maximum value for filtering spots. For example, let’s say you want to use the Glass’ effect size (a measure of Signal-to-Noise Ratio) to filter valid spots. However, you don’t know what could be a good minimum value. Therefore you can click on the dimmer valid spots and view what is their Glass’ effect size.

Finally, the Spot detection threshold function and the minimum and maximum values of the selected filtering features can be automatically tuned by SpotMAX. To do so, click on the “Start autotuning” button on the top-right of the tab.