knobby version 2

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 size.

tiling

The first row of entries correspond to the range (in um) to be covered in the x, y and z axes respectively.  The second row specifies the step size in each case.  The last entry at the bottom tells Knobby how many frames to sample at each location.

In the example above, Knobby will perform a z-stack with 40um range and 20 um steps (that is, a total of 3 locations), on an (x,y) grid of 3×3 with 200um horizontal and vertical range and steps of 100um.

As you input the desired values the table will update automatically.

Once ready, click the “Arm” checkbox (and the “Return” checkbox if you want knobby to go back to its initial position at the end) and start your acquisition.

Here is the resulting scan for the example above:

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:


% objective list
sbconfig.objectives = {'Nikon 16x','Nikon 25x'};

Now, within the Knobby panel you will see a pull down list containing the objective names:

objectives

Select the desired objective before performing a calibration.  Once the calibration is finished it will be saved.  If a calibration is not present for a given objective the calibration button will read “No Calibration” and the mouse control will be disabled. If you change the objective, simply select the corresponding entry in the pull down list to apply the new calibration (no need to restart Scanbox).

When collecting data the info structure will include the objective name and calibration data:


info =

struct with fields:

resfreq: 7930
postTriggerSamples: 5000
recordsPerBuffer: 512
bytesPerBuffer: 10240000
channels: 2
ballmotion: []
abort_bit: 0
scanbox_version: 2
scanmode: 1
config: [1×1 struct]
sz: [512 796]
otwave: []
otwave_um: []
otparam: []
otwavestyle: 1
volscan: 0
power_depth_link: 0
opto2pow: []
area_line: 1
calibration: [1×13 struct]
objective: 'Nikon 25x'
messages: {}
usernotes: ''