http://confocal-microscopy-list.275.s1.nabble.com/exorcising-spirits-from-Fluoview-tp3781225p3784179.html
The DIC prism degrades the image quite substantially on our Leica SP2. I
laser excitation is affected also. It has a quite noticable
fuzzing/doubling effect on the final image. I always recommend that for DIC
second. Have not looked at the psf, though.
> Hi Mike,
>
> We have seen a similar problem to this, particularly apparent in DIC
> transmitted light images. However, of our two FV1000 systems (located in
> different rooms), it is only readily apparent on one. This has led us to
> believe it is a vibration issue, especially as we can cause a more severe, but
> similar-looking problem by deliberately introducing a source of vibration near
> to the microscope. However, if true, we have yet to isolate the cause of the
> problem vibration.
>
> While I'm here...has anyone properly investigated the effect of the DIC
> objective prism in confocal fluorescence imaging? I had always assumed
> (rightly or wrongly) that it's presence didn't influence the PSF, but last
> week I was imaging some subresolution beads and found that, particularly on
> our IX81-based FV1000 confocals, the DIC objective prism had quite a
> pronounced effect on the psf. Specifically the psf was distorted along a
> diagonal axis and at the point of focus, the bead appeared significantly
> larger with the prism in place. The implication of this is that for confocal
> fluorescence imaging, the resolution of the microscope is reduced when the DIC
> objective prism is in place. I've also looked on our Zeiss Axiovert 200 and
> Nikon TE-2000 based systems which employ a slightly different method of DIC
> and there the effect is much less pronounced although noticeable.
>
> Simon
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:
[hidden email]] On
> Behalf Of MODEL, MICHAEL
> Sent: 07 October 2009 12:28
> To:
[hidden email]
> Subject: exorcising spirits from Fluoview
>
> I apologize if this is a second message to the list, I think the first one
> didn't go through
>
> We are having a bizzare scanning problem. Straight vertical lines in an object
> become slightly zigzagged with a period of up to 6-7 scan lines, and there
> also may be some oscillation in the intensity. The period and the magnitude of
> this periodic noise depends on the scan speed and the scan size. So far we
> (with the help of an Olympus engineer) have established that:
>
> 1. It doesn't seem to be the scanner controller or the galvo mechanism
> 2. It does not seem to be the electric power in the building
> 3. It is not a mechanical vibration
> 4. It is not a computer
> 5. It is not the cables
> 6. It is not a 60 Hz noise
>
> Sometimes connecting the scanner controller to the outlet through a long
> extension cord seemed to help which may suggest a problem with grounding, but
> as soon as we concluded that, the trick stopped working. The trouble could not
> be reproduced at the Olympus testing lab.
>
> Has anyone experienced anything similar and successfully resolved the problem?
>
> Many thanks in advance!
>
> Mike Model