Legacy Photostimulation module

A description of the Plugin can be found here: Photostimulation module

In case you want to use the Legacy Photostimulation described here, it is still possible, please contact your Scientifica representative. A description of the Standard Plugin can be found here: Photostimulation module

SciScan’s uncaging module allows users to perform point stimulation or uncaging experiments in conjunction with electrophysiology recordings and is accessed through the Plugins Menu. The uncaging module allows you to select multiple points within your field of view and use a fast laser attenuator, eg. Pockels cell, to flash these points with precise spatial and temporal resolution. Resulting activity cannot be recorded optically. i.e you cannot image the response; please use electrophysiology equipment to record responses electrically.

Uncaging Module GUI

Uncaging GUIUncaging GUI

Label Name Function
A Image/Point Tools Select between digitally zooming the image or adding point ROIs.
B Image Window Displays image from the currently selected channel.
C External Trigger Enable or disable wait for trigger before stimulation.
D Stimulation Power Relative power percentage to be applied during stimulation.
E Pre-Stim Settling Time Delay between scanning mirror movement command and Pockels cell opening to allow for the mirror to move and stabilize. This will differ with the distance between points.
F Stim Duration Duration for which laser is applied.
G Stim Interval Delay between onset of a stimulation and onset of subsequent stimulation at the same point.

If using single spot repetitions, but multiple group repetitions, the minimum stim interval is given by: (pre-stim settling time + stim duration) x number of spots.

If using multiple spot repetitions, the stim interval is the delay between the onset of one stimulation and the onset of the next. Minimum value, in this case, would be the same as stim duration.

H Group Repititions If more than one spot is selected, this number denotes the number of times the points are cycled through.
I Spot Repititions The number of stimulations per spot before moving to the next location.
J Confirm Generates output waveform for Pockels cell and scanning mirrors. Pop-up will launch with a start or cancel option before beginning.
K Close Closes the Uncaging module and returns function to the SciScan GUI.

Waveform Examples

Single Spot Repetitions

Single Spot RepititionsSingle Spot Repititions

Multiple Spot Repetitions

Multiple Spot RepititionsMultiple Spot Repititions

Use

Navigate to the channel you would like to use as a reference for the uncaging or stimulation experiment. For reference on how to access this, please see the GUI overview.

Open the Uncaging plugin through the plugin menu.

Choose the point ROI tool (A) and select the points on your image that you would like to stimulate. Press control to add multiple points.

Enter the desired parameters for Pre-Stim settling time, Stim Duration and Stim Interval.

Choose the desired power level for stimulation.

Press Confirm

A popup will launch giving you the options of “Start” or “Cancel”. Press start to begin the uncaging protocol or cancel to go back and adjust any parameters.

If “wait for external trigger” is enabled, once “Start” is pressed, the system will wait for a TTL input on the external.start.trigger.channel. For more information on triggering please see the triggering tab.

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