12. Troubleshooting
Hardware and signals
| Problem | Likely cause and fix |
|---|---|
| A channel is flat, or stuck at the top or bottom of its cell | Food touches the top electrode. Wipe it off and re-load until the trace responds |
| A whole row (P1–P12) is flat | That arena module or its cable is loose. Reseat it |
| Traces drift slowly | The arena module is not flush with its sensor board. Reseat it |
| Noisy traces | Electrical noise nearby (phones, power supplies): move them away. Arenas touching each other: separate them. Label tape used instead of masking tape: its glue can interfere with the signal |
Flies and feeding
| Problem | Likely cause and fix |
|---|---|
| Little feeding | Food set or dried: keep it at 70 °C and load a dome. Low motivation: 2–4 h dry starvation. Recent CO2: wait 24 h. Environment: 25 °C, 70 % RH |
| Food looks dry at the end of the run | With such small volumes the food always looks dry by the end of the experiment. This is normal |
| Side bias | Level the arenas; avoid uneven light |
Connection
| Problem | Likely cause and fix |
|---|---|
| Port list says “No USB device found” | The basestation is not plugged in or not powered. On Linux, add your user to the dialout group (see Installation) |
| Indicator shows Reconnecting | The USB connection dropped. The app reconnects by itself and the recording continues; dropouts are filled and shown as gaps. Check the cable and avoid USB hubs |
| An OptoPad shows Old firmware | Click Update (see Maintenance) |
| An OptoPad cannot connect | Another program is using the port, or it is the FlyPad’s port. Close the other program and choose the port labelled Arduino Mega · OptoPad |
OptoPad
| Problem | Likely cause and fix |
|---|---|
| Arm closed loop is greyed out | Start acquisition and connect at least one OptoPad; the status line says what is missing |
| LEDs do not light | Use Test in the protocol to check the LED; reseat the LED module; check the LED controller’s power |
| Too many or too few triggers | Adjust the Lick threshold using Lick signal… |
Camera and tracking
| Problem | Likely cause and fix |
|---|---|
| Find lists no camera | The camera’s driver (for Hikrobot: MVS) is not installed, or the camera is not on a USB3 port. Install the driver and plug the camera into a blue USB3 port |
| Connect says the camera is busy | Another program holds it. Connect stops MVS and left-over FlyPad Acquisition processes by itself; otherwise close the other program |
| The video frame rate is lower than set | Exposure too long (each frame needs its exposure time), or the USB link is too slow: shorten the exposure, crop the camera (Crop…) |
| The card counts dropped video frames | The PC cannot encode fast enough. Lower the frame rate, or crop the camera. FlyPad data is never affected |
| A fly is not found | It has not moved since tracking started (it is in the background): wait until it moves, or Relearn background. Check in Processing steps… which test its pixels fail |
| Tracking finds something that is not a fly | A shadow or a moved arena: Relearn background; raise Sensitivity or Min contrast; check that the arena outlines fit (Arenas… → Fit) |
| Sips with the fly on that electrode is low | The arena’s channels are swapped or wrong, or the arena is drawn in the wrong place |
Recording and analysis
| Problem | Likely cause and fix |
|---|---|
| Traces in Review look like random noise | Wrong Format. Choose 64 ch for recordings from the earlier 64-channel FlyPad |
| Analysis finds the wrong number of files | Only recordings of one experiment should be in the folder (including its subfolders). Move test recordings out |
| Results end early or look too short | Set Duration in Analysis Parameters to your assay length (360000 = 60 min). Recordings shorter than Duration are analysed up to their end and reported under Data checks |
| Conditions or labels are wrong | Conditions come from the file name, labels from Log_…txt. Fix them in the wizard’s Experiment Labels page |
Display
| Problem | Fix |
|---|---|
| Part of the window is hidden | The app sizes itself to the screen area above the taskbar. If you moved it to a smaller screen, maximize it once; it remembers the new size |
Still stuck? Contact Easy Behavior at [email protected] and attach the app’s log file:
- Windows:
%LOCALAPPDATA%\FlyPad Acquisition\logs\flypad_debug.log - Linux:
~/.local/state/FlyPad Acquisition/logs/flypad_debug.log