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Table of Contents
Troubleshooting
Try to isolate the problem to the extent possible.
| Problem | Possible Reason | Possible Fix |
|---|---|---|
| Communication error between software and hardware | Power cycle the affected hardware and restart the software | |
| One hardware element not responding | Connect to the hardware using manufacturer's software or serial terminal for Tiger controller | |
| Beams only come out of the objective if both beams are turned on | Fibers going to the wrong scanner | Exchange the fibers |
| Stage scanning cameras not getting triggered | Hardware/firmware out of date | Make sure you have Rev F XY card or later, make sure micro-mirror card has jumper on positions 11/12 of SV6 |
| Acquisition images look different than alignment (look like epi view) | Camera trigger cables swapped | Swap the camera trigger cables |
| Inconsistent communication with Tiger controller | Windows comm port problems | Update USB to comm port driver, disable USB suspend in Windows per Micro-manager recommendations |
Piezos
The piezo objective movers are the most fragile and failure-prone component of the diSPIM. See the wiki page for care and troubleshooting instructions.
Software
If you encounter bugs in the software it is best to contact the developers directly. For Micro-Manager see the wiki page for instructions.
Vibration
Some users have reported vibrations, e.g. diffraction-size beads will appear as a diagonal smear. Assuming you have the system on a floated air table, this is probably due to a combination of the vibration of the camera fan and the vibration-prone nature of ASI's original piezo objective movers. Some cameras vibrate more than others, and there can be significant variability even within the same brand/type. PCO cameras seemed to have the worst vibration initially, but in mid-2015 their internal design was modified to correct this and now all three cameras seem comparable. The exact threshold of when vibrations become noticeable depends on the experiment specifics.
There are three approaches to reduce the apparent vibration:
- Reduce camera vibration: Try swapping cameras, even with another one that is nominally identical, to see if the problem is reduced. Some cameras can be water-cooled in which case the fan can be turned off, e.g. Hamamatsu Flash4. Some cameras can turn their fans off, e.g. Andor Zyla (only recommended for bursts of acquisition, e.g. if you acquire for a few seconds every minute).
- Mechanically decouple the camera and piezo: You can mount the camera to the air table instead of the the microscope. Several groups have rigged this up themselves. In early 2016 ASI designed a universal air table mount for this purpose; it is still being prototyped but having an anxious customer would expedite things.
- Reduce susceptibility of piezo to vibrate: You can update the piezo objective mover to the version introduced early 2015 which is significantly stiffer. At the same time the scheme was changed for mounting the piezo to the SPIM arm which makes alignment significantly easier and offers some other minor benefits. The only downside to updating the piezo is that the travel range will be 200um instead of 300um. ASI performs such updates at cost, contact them for details.
