reflection-looking rings in TRANSMITTED channel

[Holly L. AARON]

Dear Confocal Experts - Something I can't seem to pinpoint:

One of my users today on an inverted Zeiss510 system is getting a ringing pattern in the transmitted channel. It looks a lot like reflection rings, but I can't figure out why there would be reflection and nothing we do seems to abate it. It is present and unchanging with different objectives, different samples (coverglass bottom slides, #1.5), different lasers. I've aligned, realigned, and unaligned the condenser, changed to different phase/DIC prisms. It remains present. I can only think it must be from one of the mirrors into the transmitted detector? Thanks for any ideas!  I'd send an image, but he's in the middle of acquisitions...

Thanks,
-Holly

[M. van de corput]

we had the same thing a while ago. It turned out that the filter block
wasn't turning propperly although everything in the system said it did.
Mariette

____________________________________________

Dr. M.P.C. Kemner-van de Corput,
____________________________________________

MGC - Dept. of Cell Biology & Genetics
Erasmus Medical Center
Dr. Molewaterplein 50, 3015 GE Rotterdam
POB 2040, 3000 CA Rotterdam, The Netherlands

Office: H-Ee751; tel: [phone]
Lab: H-Ee710; tel lab: +31 10 704.3315
tel secr: +31 10 704.3169
____________________________________________

http://www2.eur.nl/fgg/ch1/cellbiology/
http://www.thesis.kemner.biz/
____________________________________________

Op Do, 13 augustus, 2009 10:16 pm, schreef Holly Aaron:

>      Dear Confocal Experts - Something I can't seem to pinpoint:
>
>  One of my users today on an inverted Zeiss510 system is getting a ringing
> pattern in the transmitted channel. It looks a lot like reflection rings,
> but I can't figure out why there would be reflection and nothing we do
> seems to abate it. It is present and unchanging with different
> objectives, different samples (coverglass bottom slides, #1.5), different
> lasers. I've aligned, realigned, and unaligned the condenser, changed to
> different phase/DIC prisms. It remains present. I can only think it must
> be from one of the mirrors into the transmitted detector? Thanks for any
> ideas!  I'd send an image, but he's in the middle of acquisitions...
>
>  Thanks,
>  -Holly
>
>   --
>        Holly L Aaron Molecular Imaging Center Cancer Research Laboratory
> 251 Life Sciences Addition #2751 Berkeley, CA  94720-2751
> 510.642.2901 510.642.5741 FAX [hidden email]
> http://imaging.berkeley.edu

[Deanne Veronica Catmull]

Hi Holly,

 

If you look through the archives I think this has been discussed in some detail before. We have had similar trouble in the past and making sure your beam paths are set up correctly and the pinholes are properly aligned for that particular configuration would be a good starting point. If all else fails, call in your service engineer who will conduct a series of tests and re-alignment of various components.

Kind regards,

Deanne.

 

---

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Holly Aaron
Sent: Friday, 14 August 2009 6:16 AM
To: [hidden email]
Subject: reflection-looking rings in TRANSMITTED channel

 

Dear Confocal Experts - Something I can't seem to pinpoint:

One of my users today on an inverted Zeiss510 system is getting a ringing pattern in the transmitted channel. It looks a lot like reflection rings, but I can't figure out why there would be reflection and nothing we do seems to abate it. It is present and unchanging with different objectives, different samples (coverglass bottom slides, #1.5), different lasers. I've aligned, realigned, and unaligned the condenser, changed to different phase/DIC prisms. It remains present. I can only think it must be from one of the mirrors into the transmitted detector? Thanks for any ideas!  I'd send an image, but he's in the middle of acquisitions...

Thanks,
-Holly

--

Holly L Aaron

Molecular Imaging Center

Cancer Research Laboratory

251 Life Sciences Addition #2751

Berkeley, CA  94720-2751

510.642.2901

510.642.5741 FAX

[hidden email]

http://imaging.berkeley.edu

 

[Adele Vincent]

hi all

Does anyone know of a dye that is selectively taken up by astrocytes in culture that would be compatible with calcium imaging (Fluo-4 or Calcium Orange) and not require post-imaging immunostaining?

SR-101 works in vivo, but presumably not in vitro. Has anyone tried it? Are there any other options?

thanks
Adele