Not long ago I mentioned Scanbox’s new ability to sample on a surface. Now you can access this new feature in the GUI by navigating to the Surface Sampling panel.
Surface sampling allows you to link lines of the resonant scan to depths determined by the optotune setting. In other words, it allows you to sample on a surface along the galvo axis (the vertical axis in Scanbox). Of course, limits are imposed by the range of the optotune and its dynamical response.
Below is an example of how the process works. Here, I am imaging a slide of pollen grains that is tilted along the vertical axis.
Because the slide is tilted, different settings of the optotune bring the grains in different lines into focus, as shown at the beginning of the video.
To compensate for the tilt we can establish a link between lines in the scan and depth. To do this, change the optotune setting while focusing, then hit the Link button, and then click on the grains that are in focus. In this example, I repeat this a handful (3) of times.
Now, when the Enable button is clicked, Scanbox interpolates a value of the optotune for each line given the established links and uploads the resulting values to the Scanbox firmware. When we image the slide with the link active we see all the grains in focus. In other words, Scanbox is now sampling on a slanted surface and compensating for the tilt of the slide.
This is a useful feature to use when compensating for the curvature of a structure that is being imaged or tilting the imaging plane without physically tilting the objective.
Try it and let me know if you run into any problems.
The use of this feature requires an update of Scanbox and the firmware to version 4.0.
Scanbox 