Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Another possibility is that the pinhole is terribly misaligned. But it does
not explain why you just see the artifacts from those samples. Another thing
you may try is to remove the DIC prism which could cause some problems.

Good luck,

Xuejun

-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Guy Cox
Sent: Wednesday, June 01, 2005 11:57 AM
To: [log in to unmask]
Subject: Re: FW: 'Double ines artefact' on Zeiss LSM

Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

If you have a reflective layer beneath your sample
(ie an interface between different refractive indices)
it's entirely possible to get a complete mirrored
second image of your sample formed by rays reflected
from the layer (sketch it on a piece of paper and
you'll see what I mean).

                                        Guy


> Search the CONFOCAL archive at
> http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
> A colleague of mine, has just  found something really odd
> with Z scans on the inverted Zeiss LSM confocal. When they do a
> series of confocal sections through polarized renal cells there
> appears to be a double band in the orthogonal slice.  The stain is
> WGA-FITC,
> and it doesn't appear to be any more intense than other staining seen
> before.  The cells were grown on filters, which do fluoresce a little but
> the double line seen, we are assured is not the filter! Has anyone ever
> seen
> /heard of this before?
> Could it be some sort of a reflection?
>
> Thanks
>
> Michelle
>
>
> Dr Michelle Peckham
> Senior Lecturer
> Postgraduate Admissions Tutor
> School of Biomedical Sciences
> University of Leeds
> Worsley Building
> Leeds
> LS2 9JT
> UK
> [log in to unmask]
> www.cell-motility.com
> 0113 343 4348 (phone)
> 0113 343 4228 (fax)
>
>
>
>
>
>


--
Associate Professor Guy Cox
Electron Microscope Unit,
University of Sydney,
NSW 2006, Australia

Phone:+61 2 9351 3176    Fax:+61 2 9351 7682
http://www.guycox.net


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