Dear Shawn, sorry for my late response!
The answer to first question is shown in “20211116-10X-Z0.6-AllLasers.jpg” (all examples are uploaded in the next post), where a sample was scanned with four lasers (this is autofluorescent signal). I see no principal difference between the channels, but my previous calculations indicate that the shrinking is a little bit higher at lower wavelength. To do this calculation, I located an object and scanned it in two positions - to be at the top of the scannning area and at the bottom. Then I measured the vertical size of the object and obtained two values (sizes): the size at bottom was a little bit higher than at the top (so the image is vetrically shrinked with non-linear dependence). This may arise from the more higher distortion at the bottom of the scanning area. As I described in my previous posts, this is a function of objective magnification (higher X covers less physical area, so the distortion is less) and a laser line (the higher energy wavelength will refract higher and the distortion is higher as well).
The answer to last question is YES, my microscope is equipped with DIC analyzer under the objective turret, but I use empty slot and it does not block any part of the light pathway. I also checked all the positions of all optical things and did not found any improper installation or positioning, which could affect this.
I have checked again the LSM modules. Unfortunately, I cannot conclude anything. First, I run CanCheck and found that there is the only problem - the potential conflicts of motorized control for spectral detector components (they are called as SDAL/SDAS/SDPINL/SDPINS/SDPL/SDPS/SDWL/SDWS). Here the explanation of the abbreviations:
SD = Spectral Detector (first two characters)
L = Long wavelength (last character)
S = Short wavelength (last character)
P = prism (third character, if not PIN)
W = wedge (third character)
PIN = pin (third-to-fifth letters)
Please see image “008.jpg” as an example, the red marks show this problem. Actually all these modules say they are in potential conflict with counterparts (e.g. SDAL against SDAS). When I move all to the “Safe position” (using CanCheck), the red marks disappear. However, when I then run a test for, say, SDAL, it passes the test successfully and moves to a new position, but the counterpart (SDAS) now again has a red mark for SDAL. Nevertheless, motorized components of spectral detector all passed the tests run by CanCheck or System Maintenance Tool (SMT, please see image “SMT-MotorTest.jpg”). I also compared the results in CanCheck reports obtained now and an year before, no differences were found between the entries related to the spectral detector (as well as other components).
As the image is out-of-center and shrinked, I am unable to run “Resolution Test”, “Scan Field Test” or “Scanner Calibration” using calibration objective by System Maintenance Tool.
To summarize, I am so wondering how much is the extent of image shrinking, now I do not believe that the problem comes solely from the optics. But I am in a dead end now and don’t know how to proceed…
PS About the contracts between Zeiss and a customer for servicing - in my location there is somewhat another regulation, which anyway assumes that the repairment result has to be approved by a customer. In our case last year, it was approved by us before the problem with Z-movement of objective turret next day, and then Zeiss repaired the problem within the actual contract with no additional costs. Indeed, my Institution takes 10-20% of the grants (it depends on the grantor), but this amount is not so much and it is used for many other purposes, not only for repairment. Also, while there is a Zeiss representative just in my city, the costs are still high because of the currency rate (they work with Euro). I am from Yekaterinburg, Russia.