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ILDA functions for FRAP, TIRF, FLIM modules

  1. If you have an additional module such as a FRAP, TIRF or FLIM module, ILDA fuctions will need to be configured. You can continue setting up the recipe by clicking Next.
  2. In the Ilda Functions tab, select your device connected to the Ilda connector (only available for the XL box type).
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  3. All Ilda functions and all parameters in each function are automatically loaded. Check if all functions and parameters are correct, then click on the Laser tab.
  4. In the Laser tab, you will find a table where you must specify the shutter and power Sub Devices for each laser line. Select the Sub Devices by clicking on the drop-down menu in each column.
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  5. Then click on the FRAP Mode tab.
  6. Select Active Frap Mode to access this option in the interface (if the box is unchecked, no FRAP parameters appear in the interface):
    1. Select the sub-device that determines the frap density and the scanRate.
    2. Specify the density index, which must match the FRAP diameter value set for the sub-device in the Device tab.
    3. Select the Sub Device that determines the Fire Preview.
    4. Active Channel Elements shutter for FRAP (Allows to indicate which channel is a FRAP channel).
    5. Add the discriminant for the frap calibration like Objective, filter cube.
    6. Add camera calibration.
      Note: If you have several cameras indicate if you want to use the same calibration for all cameras or a different one. If you want to use the same calibration indicate the same number in the calibration column for all cameras
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  7. When it’s done, then go to the TIRF Mode tab.
  8. Select Active TIRF Mode to access this option in the interface (if the box is unchecked, no TIRF parameters appear in the interface):
    1. Select the Sub Device assigned to the TIRF density.
    2. Specify the density index which need to be the same number than in the subdevice TIRF diameter in the Device tab.
    3. Select the Sub Device responsible for the Angle, Angle Length and the radius. Those parameters will help you to configure as you want a TIRF experiment.
    4. Active Channel Elements for TIRF (allows to indicate which channel is a TIRF channel).
    5. Add discriminant for the TIRF calibration like Objective, filter cube, lasers.
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  9. If you have a lightsheet system, go to the ROI Mode tab (for ROI scanning).
    1. Select Active ROI Mode to access to this option in the interface.
    2. As with FRAP, select the sub-device that determines the density and the scanRate.
    3. Specify the density index which need to be the same number than in the subdevice frap diameter in the Device tab.
    4. Add active channel elements ( if you activate one element in the channel that means you are in the ROI mode).
    5. Add discriminant.
    6. Add ROI.
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  10. The Illumination Mode is used to add a virtual device to select the illumination mode (e.g. if you have a multimodal system with FRAP, TIRF, Spinning Disk).
    1. Select Active Illumination Mode to access to this option in the interface.
    2. As with FRAP, select the Sub Device that determines the density and the scanRate.
    3. Specify the density index which must to be the same number than in the Sub Device frap diameter in the Device tab.
    4. Add active channel elements.
    5. Add discriminant.
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  11. Click on Next to proceed to the last step, which is the FLIM configuration.
    Fastpath: If you don’t have the FLIM module, you can directly click on Create Recipes.
    1. Add all devices responsible for the Delay.
    2. Add discriminant.
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  12. Click on Create Recipes. Your recipe is created. You can now finalize your I.S. configuration.