Operation

Ephus

  • Ephus should be configured to save the xsg file. This can be loaded as a mat file that contains the state of Ephus during the acquisition. It contains a structure called header which contains the pulse info and gain along with the actual stimulus trace​

Configuration hotswitches

  • Save configuration settings for program windows regarding recording and stimulation settings

  • See:

    • Programs → hotswitch
    • Programs → hotswitch: hs_config

  • Located in C:/Rig/Ephus 2013b/hotswitches

Looping

  1. Programs → LoopGui
  2. Set number of iterations
  3. Set interval time between iterations
  4. Ensure stimulator and acquirer are set to external and use PFI9 for triggering

ScanImage

  • 145×145 μm images at 256×256 pixel resolution at an effective frame rate of 5 Hz

Maintaining cells in field-of-view

  • Use screen annotation software (such as gInk) to annotate cells so that they may be kept in focus in the field-of-view throughout the experiment.

Looping (cycles)

Cycles allow multiple iterations with defined delays between acquisitions. Cycles are saved in: C:\Rig\Scanimage5-3\cycles

Setup procedure:

  1. View → Cycle Controls to open cycle configuration (CYCLE-MODE CONTROLS)
  2. Check Cycle mode on
  3. Load existing cycle or save new cycle configuration

Configuring iterations:

Multiple iterations can exist for each cycle, with each iteration preceded by a defined Iteration Delay.

Example: 1 cycle with 4 iterations, each with 120s delay

  1. Click >> for cycle iteration data
  2. Cycles done: displays X of 1
  3. Add 4 iterations
  4. Enter 131 in Iteration delay field and press Enter
    • Calculation: Frame collection time (~11s for 50 frames at 5 Hz) + desired delay (120s) = 131s total
  5. Check Restore original cfg
  6. Check Auto-reset iteration/count
  7. Click CYCLE to begin cycle

Note: There is a timing delay of ~97-99 ms (occasionally up to 700 ms) between the ScanImage acquisition start message (e.g., "Starting '1 x zoom 5 Hz' at 6/13/2018 12:33:16.136") and the acquisitionStarted timestamp in the digtrig_stim user function.