Intrinsic & epi-fluorescence imaging using the port camera

Some colleagues have asked about the possibility of integrating imaging acquisition from the port camera to do intrinsic and/or epi-fluorescence imaging ahead of targeting a region of interest with two-photon experiments in Scanbox. We have now added this option to Scanbox. To use this  option you will need to wire the I2C port on the…

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Sampling on a Surface

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…

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Tiling with Knobby

Knobby scheduler allows the user to move the microscope by specifying a list of relative changes in position at given frame numbers.  One quick way to fill up the table is provided by a set of text fields in the knobby panel.  This allows one to perform (x,y,z) tiling with a given range and step…

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Processing Volumetric Data

After collecting volumetric data with the Optotune you can now process your data as follows. First, use sbxsplit() to generate separate data files for each “slice” of the optotune. For example, if I have a data file gn7_008_001.sbx collected some data with an optotune waveform having a period of 5 the command will generate a…

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Spatial Calibration for Multiple Objectives

Multiple users of the scope may be running projects that require different types of objectives.  How to keep a spatial calibration for each and switch between them when necessary? Scanbox now includes an “objective” configuration variable — a cell array of strings each with the name of a different objective.  Right now I have: Now,…

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Visualizing Individual Slices during Volumetic Imaging

During volumetric imaging, Scanbox displays all images as they are acquired. This can be inconvenient if we are trying to visually assess the activity of neurons within any one optical slice.  One solution is to display the images separately in a montage by means of a Scanbox plug-in.  However, it would be better to have…

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Automatic Control of Laser Power

A new checkbox within the Laser panel (labeled AGC) allows you to turn an automatic control of laser power on and off. When AGC is on, Scanbox checks the distribution of pixel values on the image every T seconds, and increases or decreases the laser power by a certain factor if the fraction of pixels…

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Automatic Optotune Calibration

We previously explained how to calibrate the optotune manually.  With the introduction of Knobby 2, we are able to make this process automatic.  You will now find a ‘Calibration’ button in the Optotune panel.  To use it, do the following: Set the optotune slider to its lowest value (slide all the way down) Bring some pollen…

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Click-and-Center with Knobby Mouse Control

If you have been making use of Knobby’s spatial calibration button, you can now move on to interacting with Knobby remotely. The new Knobby scheduler panel looks a bit expanded.  The speed (coarse/fine/super-fine), the mode (normal/rotate), and zero (XYZ/XYZA) buttons should be self explanatory. They do exactly the same as if you were to be…

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Spatial Calibration and return to origin

We added two new features to the knobby scheduler panel, as shown below by the arrows. One (left arrow) allows knobby to perform an automatic spatial calibration of the system, measuring the (x,y) size of pixels at all magnifications.  To perform this calculation do the following: Focus on some pollen grains.  Make sure one of the…

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