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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| Stage scanning cameras not getting triggered | Hardware/firmware out of date | Make sure you have Rev F XY card or later, make sure micro-mirror card has jumper on positions 11/12 of SV6 |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 (NOT universal version), 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-ability” of the piezo objective movers. 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; there can be significant variability even within the same brand/type of camera. 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). PCO Edge cameras seemed to have the worst vibration initially, but in mid-2015 their internal design was modified to correct this and now all three major camera brands seem roughly comparable.
- Mechanically decouple the camera and piezo: You can mount the camera to the air table instead of the the microscope. As of mid-2016 ASI offers a universal air table mount for this purpose, and several groups have rigged this up themselves. There is some evidence that the CDZ-R block used on RAMM-mounted diSPIM reduces the vibration compared with the CDZ-1000 used when the diSPIM is mounted on other inverted microscopes.
- Reduce susceptibility of piezo to vibrate: If you have an 2014 or older ASI piezo objective mover you can update to the version introduced early 2015 which is significantly stiffer and hence less prone to vibration. At the same time the mounting scheme was changed which makes alignment significantly easier and offers other minor benefits. The only downside to updating the piezo is that the travel range will be 150um instead of 300um. ASI performs such updates at cost, contact them for details. If you are only doing stage scanning the piezos can be eliminated entirely.
