Protocol

Recording S. mansoni miracidia behavior in custom arenas using the InVision imaging system, an assay procedure to evaluate responsiveness to cues of choice. This protocol explains how to prepare the InVision system. A follow-up protocol explains how to run behavioral assays.

Setting Up InVision:

1. Plug in the power supply chord (black chord) into the outlet on the wall, and camera chords (blue chords) into the “camera input” on the InVision computer.
2. Turn on the monitor/computer by pressing the power button.
3. After a couple minutes, the monitor will turn on. Select “lab” as the sign-in option. Type in the password and wait for the desktop screen to load.

4. Open the “Motif” application by clicking the blue and white “M” icon at the top right of the screen. Select “Open Motif” from the drop-down menu.
5. Once the application opens, connect both cameras under the “Local Cameras” widget. Select the blue “Connect” button, one at a time, next to each camera. The left camera is named “24568709” and the right camera is “24568744”. The buttons will turn from blue to gray and two windows (one for each camera) will pop up below when the cameras are connected.

6. Make sure you are on the “Record” tab, below the “Local Cameras” widget. Set the camera exposure on each camera to 750.0 µs. Give the recording a title (”filename”) and a description. Template and image below.
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METADATA TEMPLATE: Filename (example): 20240115_a01_RVH where 20240115 is the date, a01 is the arena index, and RVH are the initials of the person doing the experiment. The system automatically saves files with the date and camera # added to the filename. Description (example): “First replicate, double agar arena design, 20 microliter aliquot of miracidia added, 0.5% SCW and 0.5% APW agar cubes used, SCW agar cube is on the right (camera: 24568744), arena filled with APW

7. Navigate to the “Configure” tab, right next to the “Record” tab. Type in the “Recording Duration” as “01:00:00,” where the numbers represent hours:minutes:seconds. Set the “Recording Codec” to hq-fast, and set the “Max Framerate” to 8.0.

8. Click the “IQ/Schedule” tab. Set LED to 10.0 and both fans to 0.75 by dragging the blue slider or by typing in the values manually.
9. Navigate back to the “Record” tab and once ready to record, select the blue “Record” button above the metadata (filename and description). After this, the system will record for 1 hour and then you will need to move the videos onto Wheeler Lab network attached storage server. This is described below as the last step(s) in the “Performing Behavioral Assays” section.

10. After the last replicate of the day, navigate to the “Move/Copy” tab of the Motif software. Click the Move All button to transfer the recordings from the InVision hard drive to the the Wheeler Lab server (data.wheelerlab.bio). The data on the hard drive will automatically be deleted after the transfer is complete.