Troubleshooting and Tips for Zeiss Microscopes

Note on cleaning: (objective lenses, camera, filters for fluorescence on
slider.)
Please try to avoid doing this.
a) The camera should NEVER be cleaned - it needs to be in a cleanroom to be cleaned.
b) Objective lenses subject to repeated cleaning can lead to loss of performance
c) When sliding between filters with fluorescence be CAREFUL not to touch with your fingers. It is not good to clean these filters, and if the dichroic is damaged, the cube is useless.

Problem: Condenser does not hold position and slides down.

Suggestion: The screw holding the condenser may be loose. This is a side screw on the left side opposite the knob where we focus the condenser. A small allen wrench is used to tighten it. When the heat is turned on, over time, the grease will loosen and we need to tighten it periodically.

Problem: Dust on digital camera lens; particles show up in images.

Remove the camera from the microscope. Use compressed air to blow the dust off. If that doesn't work, a small amount of glass cleaner on some lens paper should do the trick. Avoid excessive pressure with dry lens paper.

Fluorescence Lamp

If you ever adjust the fluorescence alignment, make sure that you never pull on the collector lens knob (this is the big black knob on the
lamp housing) -- it has to fit into a groove in the lens collector, and if it falls out, then we lose our focusing capability.

Problem: Media spilled on motorized stage or objective

Please be careful and at the end of your experiment check to see if there have been any leaks. It is important to carefully clean the stage and microscope to prevent rusting and to keep everything working properly. Use cottonswabs for the stage, but use only proper lens paper for the objective. Please see Joyce if you have any questions. It is better to ask if you have doubts.

Thanks! A simple check at the start and end of an experiment will help greatly in microscope maintenance.

Metamorph

In general, if there are any problems with metamorph -- even if it is that it doesn't recognize the key -- Joyce would like to know. Neal Gliksman is
very helpful, and a lot of the issues that have been brought up recently have been solved.

Email files

Email files that are being generated randomly in your image gathering folders was due to the fact that the email function in the MM drop-in manager was installed. The only reason to have this installed is if you want to send MM images from the MM application as an attachment via email. Joyce saw no reason for this so she removed the email drop-in.

Image file saving

Randomly, it appeared that some files were saved as 8-bit images. It appears that some of the journals in the "define camera acquisition settings" had the 8-bit image transfer option checked. Please make sure when you write a journal that you check to see that all the options are correct. There was the fitc journal that appeared to get 'subtract background' checked, and this seemed to happen randomly. Thus be careful and check before you gather your images with journals. Don't assume that the journal is correct.

Try to use the Acquire function under the Acquire menu -- this is more versatile in the settings for gathering images.

Princeton Instruments camera driver

If you have a problem with Princeton Instruments camera driver being recognized by MM, you may have to go under programs, MetaImaging 4.5 and
open the Video Manager. Click on configure (when PI is highlighted) and then click on Autoconfigure. This seems to work to let MM know to communicate with the camera.

Version

Joyce updated Metamorph to version 4.5r6 in October (the one next to the microscope).

There should be no problems. However, if there are problems, and you would like to convert back to 4.5r2, send an email out first to discuss. It is
very easy to go back and forth between the two. Go into C:/MM and open up mmupdater.exe -- when it asks you for the updater, you can click on either the zip file for 4.5.2 or 4.5.6.

Problem: Uneven brightness or loss of light intensity in brightfield.

Suggestion: Any filters in the optical path will decrease the light. Above the condenser, there are two filters
that can be swung into and out of the optical path. The top filter (just below the ring where you close down the condenser aperture for Kohler illumination) contains a blue filter which cuts down on quite a bit of the light. If you remove this filter, you may find that the cells are not happy because they may be exposed to too much light. The filter just below this is the polarizer, and this can be pushed out unless you are doing DIC. The other source which you would be losing light is the slider (which contains the fluorescence filters below the microscope objective nosepiece).

Reason why slider between camera and eyepiece broke:
a) Abuse to the slider. Surprisingly, simply placing your palm on the slider and pushing is enough to do damage the mechanism.
b) The prism is cemented onto the piece that moves with the slider which hits a stop screw when pushed. The slightest pressure and repeated use causes the cement to break loose.
c) Therefore, the prism will be come misaligned, and the slider will be either tight or loose depending on how the prism moves.
IF AT ANY TIME, ANY PARTS ARE LOOSE, PLEASE CONTACT DOUG FISHKIND, OUR ZEISS REP IMMEDIATELY. Contact info should be on website.

Problem: Thermocouple is loose

Please do not use tape to attach the thermocouple -- the adhesive on the tape melts onto the microscope. Use copper wire to secure the thermocouple to the microscope condenser.

Problem: Half of the field of view is obstructed.

Suggestion: Check the slider on the left side which cuts off the light from the eyepiece. It sticks sometimes.