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Projector Plugin

Revision as of 12:06, 25 September 2013 by Arthuredelstein (Talk | contribs)

The Projector plugin provides a single graphical user interface for controlling SLMs (spatial light modulators), such as DMDs (digital mirror devices), LCOS (liquid crystal on silicon) modulators or even digital computer projectors, and also for controlling steerable-mirror (e.g. galvo-based) phototargeting devices.

The Projector plugin's interface shows three tabs: Point and Shoot, ROIs, and Setup. How to operate each tab is described in detail below.

Setup Tab

Plugin projector-setup.png

The Setup tab allows you to test the phototargeting device and perform initial calibration. Press On and Off buttons to turn the light on and off (in the case of a pulsed laser such as the MicroPoint, the On button will only turn the light on briefly). Press the "Show center spot" button to illuminate the center of the phototargeting device's range. This Center Spot should be visible in the camera's field of view -- if it isn't, the physical alignment of the camera and/or phototargeter will need to be adjusted.

The Calibrate button starts an automated calibration procedure, where the phototargeter's internal coordinates are mapped to the pixel coordinates to the camera's pixel coordinates. This calibration allows the Projector plugin to convert mouse clicks or ROIs drawn on the live camera image into precisely directed phototargeting events.

The calibration procedure starts by illuminating a few spots near the center of the phototargeter range. This gives the Projector plugin an estimate of the orientation and location of the camera field of view relative to the phototargeter range. Then, points in a square lattice are illuminated one by one, and imaged on the camera. The Projector plugin then uses the lattice measurements to compute a mapping of each pixel to phototargeter coordinates for future phototargeting. Mappings for each camera and phototargeting deviced are stored and will be remembered even after Micro-Manager is restarted.


Setting Description
On / Off Toggles device illumination.
Calibrate Creates calibration map of phototargeting device range to the camera pixel position.
Show center spot Illuminate center of SLM or galvo device.
Phototargeting channel Toggles to specified channel during phototargeting. You don't need to set this unless some hardware needs to be moved during imaging, like a dichroic and/or a shutter. If you don't see your channel listed, you must set the Core-ChannelGroup in the Property Browser to the Group containing this channel.

Point and Shoot

Plugin projector-point and shoot.png

Point and Shoot lets you phototarget a small region instantaneously, by clicking on a live image of the specimen. Galvo-based devices fire a single shot, while SLM devices will turn on in a small region around the clicked pixel. The procedure is very simple:

  1. Under the Point and Shoot tab, set "Point and shoot mode" to "On."
  2. Turn on live mode.
  3. Hold down the Control key (on the keyboard) and click anywhere in the live image window. You should immediately see a laser spot appear for a time at the clicked location.

ROIs tab

Plugin projector-rois.png


The Regions of Interest (ROIs) tab lets you specify regions on the image to be phototargeted. Phototargeting of an ROI can be carried out instantly, or programmed into a Multi-Dimensional Acquisition.

  1. Before running an ROI experiment, the calibration (described above under "Setup Tab") must already have been carried out. Over time, the calibration may drift, so repeat calibration as needed. It's also useful to try "Point and Shoot" first, to make sure the phototargeting hardware is working and the calibration is correct.

The procedure for illuminating a single ROI is as follows:

  1. Select one of the ROI tools on the ImageJ toolbar. Possible ROIs tools can include Rectangular, Oval, Polygon, Freehand, or Point. (Note that some devices, such as the MicroPoint, currently only support Point targeting.)
  2. Press "Set ROI(s)". The ROI will be uploaded to the phototargeting device.
  3. Choose a value of 1 or more for "Loop". If Loop is greater than 1, then for every phototargeting event, the phototargeter will loop through the set of ROIs multiple times.
  4. Turn on Live mode (on the main Micro-Manager window).
  5. Press "Run Rois now!" The ROI should be illuminated (rastered in the case of a galvo-based device).

It's also possible to run multiple ROIs:

  1. Click the ROI Manager button to display ImageJ's ROI Manager.
  2. Draw an ROI, and press "Add." The ROI will be included in the list.
  3. Press "Set ROI(s)", turn on Live mode, and then "Run Rois now!"

Pressing "Run ROIs now!" is a good way to phototarget the specimen during a Multi-Dimensional Acquisition. Just start the MDA, wait for the right moment, and then hit "Run Rois now!" to cause phototargeting to start, while acquisition continues simultaneously.

You can also run ROIs at pre-determined time points in an acquisition sequence, however:

  1. Check Run ROIs in Multi-Dimensional Acquisition
  2. Enter a "Start Frame" index -- that is, the first frame at which phototargeting should commence.
  3. If further phototargeting events are desired, check "Repeat every" and specify how many frames should elapse before the next phototargeting event should occur (after the first).
Setting Description
Set ROI(s) Program the device with ROI(s) drawn on the selected image.
ROI Manager To add multiple 2D regions on the image, one has to use the ImageJ ROI Manager. This button is just a shortcut to that ImageJ menu.
Loop Number of times to fire ROIs. This setting differs from the device's Repetitions property by looping after visiting each region.
Spot dwell time (Does not affect pulse laser) Stops photo illumination after set time.
Run ROIs now! Fires ROIs drawn on current image.
Run ROIs in Multi-Dimensional Acquisition Fires phototargeting devices in parallel with MDA (does not affect image capture rate).
Start Frame Timepoint in MDA to fire sequence.
Repeat every __ frames Timepoint interval in MDA to again fire sequence until MDA completes or device turned off in Setup tab.


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