Dirty 20x dipping objective on 2-photon system

[Oliver Biehlmaier-2]

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Dear fellow microscopists,

We have very strange dirt / crystals -image on: https://imgur.com/a/PjOnsSC - on a Zeiss dipping objective which we are using on our 2ph-tomography system.

We tried to clean it with Ethanol 70%, Ethanol 100%, Chloroform, Ether/Ethanol 30%/70% with zero improvement of the situation.
Thus we guess that the dirt is inside the objective, very much likely salt crystals originating from the buffer solution that the objective is dipping in.
Have you ever seen something like this? Is it possible that the sealing of the objective is broken after approx. 3 years of dipping and as a consequence these crystals could build up on the rear side of the front lens?

All the best,
Oliver



Oliver Biehlmaier, PhD | Head of Imaging Core Facility  | Biozentrum, University of Basel | Klingelbergstrasse 50/70 | CH-4056 Basel
Phone: [phone] | Email: [hidden email]<mailto:[hidden email]> | www.biozentrum.unibas.ch/imcf<http://www.biozentrum.unibas.ch/imcf> | www.microscopynetwork.unibas.ch<http://www.microscopynetwork.unibas.ch>

[Anne Vaahtokari]

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Hi Oliver,



Have you tried to dissolve the crystals in water?



Regards,

Anne



Sent from Mail<https://go.microsoft.com/fwlink/?LinkId=550986> for Windows 10



________________________________
From: Confocal Microscopy List <[hidden email]> on behalf of Oliver Biehlmaier <[hidden email]>
Sent: Monday, May 28, 2018 8:38:53 AM
To: [hidden email]
Subject: Dirty 20x dipping objective on 2-photon system

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Dear fellow microscopists,

We have very strange dirt / crystals -image on: https://imgur.com/a/PjOnsSC - on a Zeiss dipping objective which we are using on our 2ph-tomography system.

We tried to clean it with Ethanol 70%, Ethanol 100%, Chloroform, Ether/Ethanol 30%/70% with zero improvement of the situation.
Thus we guess that the dirt is inside the objective, very much likely salt crystals originating from the buffer solution that the objective is dipping in.
Have you ever seen something like this? Is it possible that the sealing of the objective is broken after approx. 3 years of dipping and as a consequence these crystals could build up on the rear side of the front lens?

All the best,
Oliver



Oliver Biehlmaier, PhD | Head of Imaging Core Facility  | Biozentrum, University of Basel | Klingelbergstrasse 50/70 | CH-4056 Basel
Phone: [phone] | Email: [hidden email]<mailto:[hidden email]> | www.biozentrum.unibas.ch/imcf<http://www.biozentrum.unibas.ch/imcf<http://www.biozentrum.unibas.ch/imcf<http://www.biozentrum.unibas.ch/imcf>> | www.microscopynetwork.unibas.ch<http://www.microscopynetwork.unibas.ch<http://www.microscopynetwork.unibas.ch<http://www.microscopynetwork.unibas.ch>>

[Roland Nitschke]

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Dear Oliver,

looks really impressive, what ever you do, even resolving the crystals
will not help if you manage to reach the compartment where they are, you
will be left with other new carbage after drying and the crystal will
come back again as there must be a hole somewhere in the glue. Also the
whole lens looks quite turbid.

I would send in the lens to Zeiss for repair, it should not be to
expensive as the front lens does not seem to be damaged. They will take
it apart, clean and realign everything. If it is the 20x/1.0 Plan
Apo-Chromat I would expect around 2000 €, if it is the one with the
Corr-ring it will be more. Still they might replace the frontlens, if it
turns out it has some scratches currently not visible due to the crystals.

We have several of the dipping lenses from ZEISS, also older ones and I
never have seen that, but 2 times on other objectives stuff appeared
inside and was unreachable and therefore uncleanable.

Best

Roland



***********************************************************
*********************************************************** Dr. Roland
Nitschke Albert-Ludwigs University Freiburg Life Imaging Center (LIC) in
ZBSA Microscopy and Image Analysis Platform (MIAP) Habsburgerstr.49
79104 Freiburg Germany e-mail: [hidden email]
<mailto:[hidden email]> phone: [phone] fax:
[phone] web LIC:
https://miap.eu/miap-units/life-imaging-center-lic/ web MIAP:
https://miap.eu <https://miap.eu/>


Am 28.05.2018 um 16:38 schrieb Oliver Biehlmaier:

> *****
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> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear fellow microscopists,
>
> We have very strange dirt / crystals -image on: https://imgur.com/a/PjOnsSC - on a Zeiss dipping objective which we are using on our 2ph-tomography system.
>
> We tried to clean it with Ethanol 70%, Ethanol 100%, Chloroform, Ether/Ethanol 30%/70% with zero improvement of the situation.
> Thus we guess that the dirt is inside the objective, very much likely salt crystals originating from the buffer solution that the objective is dipping in.
> Have you ever seen something like this? Is it possible that the sealing of the objective is broken after approx. 3 years of dipping and as a consequence these crystals could build up on the rear side of the front lens?
>
> All the best,
> Oliver
>
>
>
> Oliver Biehlmaier, PhD | Head of Imaging Core Facility  | Biozentrum, University of Basel | Klingelbergstrasse 50/70 | CH-4056 Basel
> Phone: [phone] | Email: [hidden email]<mailto:[hidden email]> | www.biozentrum.unibas.ch/imcf<http://www.biozentrum.unibas.ch/imcf> | www.microscopynetwork.unibas.ch<http://www.microscopynetwork.unibas.ch>

[Mark Cannell-2]

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From the image I can't make out if the 'crystals' are on the front or the back of the lens... I can't help wondering idf these are not crystals at all but erosion of the front lens surface possibly inder a coating. This could happen in fuorite lenses in the presence of moisture I think.

Good luck but it looks pretty bad.

Mark B. Cannell. Ph.D. FRSNZ FISHR
Department of Physiology, Pharmacology & Neuroscience
School of Medical Sciences
University Walk
Bristol BS8 1TD
 
[hidden email]
 
 

On 28/05/18, 3:39 PM, "Confocal Microscopy List on behalf of Oliver Biehlmaier" <[hidden email] on behalf of [hidden email]> wrote:

    *****
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    Post images on http://www.imgur.com and include the link in your posting.
    *****
   
    Dear fellow microscopists,
   
    We have very strange dirt / crystals -image on: https://imgur.com/a/PjOnsSC - on a Zeiss dipping objective which we are using on our 2ph-tomography system.
   
    We tried to clean it with Ethanol 70%, Ethanol 100%, Chloroform, Ether/Ethanol 30%/70% with zero improvement of the situation.
    Thus we guess that the dirt is inside the objective, very much likely salt crystals originating from the buffer solution that the objective is dipping in.
    Have you ever seen something like this? Is it possible that the sealing of the objective is broken after approx. 3 years of dipping and as a consequence these crystals could build up on the rear side of the front lens?
   
    All the best,
    Oliver
   
   
   
    Oliver Biehlmaier, PhD | Head of Imaging Core Facility  | Biozentrum, University of Basel | Klingelbergstrasse 50/70 | CH-4056 Basel
    Phone: [phone] | Email: [hidden email]<mailto:[hidden email]> | www.biozentrum.unibas.ch/imcf<http://www.biozentrum.unibas.ch/imcf> | www.microscopynetwork.unibas.ch<http://www.microscopynetwork.unibas.ch>
   

[Craig Brideau]

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Coatings are more likely to flake off than form feather patterns like that.
The patterns imply salt growth. All you can do is soak the lens tip out in
ultra pure water to try to dissolve the salt. The fact that it is in there
to begin with, however, means that the lens seal is compromised and needs
to be repaired. Even if you soak out the salt (which will take you a while
or be impractical if the seal leak is very small) you will still probably
have some residue, so the lens actually needs to be disassembled, cleaned,
reassembled, and resealed.

Craig

On Mon, May 28, 2018 at 9:53 AM Mark Cannell <[hidden email]>
wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> From the image I can't make out if the 'crystals' are on the front or the
> back of the lens... I can't help wondering idf these are not crystals at
> all but erosion of the front lens surface possibly inder a coating. This
> could happen in fuorite lenses in the presence of moisture I think.
>
> Good luck but it looks pretty bad.
>
> Mark B. Cannell. Ph.D. FRSNZ FISHR
> Department of Physiology, Pharmacology & Neuroscience
> School of Medical Sciences
> University Walk
> Bristol BS8 1TD
>
> [hidden email]
>
>
>
> On 28/05/18, 3:39 PM, "Confocal Microscopy List on behalf of Oliver
> Biehlmaier" <[hidden email] on behalf of
> [hidden email]> wrote:
>
>     *****
>     To join, leave or search the confocal microscopy listserv, go to:
>     http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>     Post images on http://www.imgur.com and include the link in your
> posting.
>     *****
>
>     Dear fellow microscopists,
>
>     We have very strange dirt / crystals -image on:
> https://imgur.com/a/PjOnsSC - on a Zeiss dipping objective which we are
> using on our 2ph-tomography system.
>
>     We tried to clean it with Ethanol 70%, Ethanol 100%, Chloroform,
> Ether/Ethanol 30%/70% with zero improvement of the situation.
>     Thus we guess that the dirt is inside the objective, very much likely
> salt crystals originating from the buffer solution that the objective is
> dipping in.
>     Have you ever seen something like this? Is it possible that the
> sealing of the objective is broken after approx. 3 years of dipping and as
> a consequence these crystals could build up on the rear side of the front
> lens?
>
>     All the best,
>     Oliver
>
>
>
>     Oliver Biehlmaier, PhD | Head of Imaging Core Facility  | Biozentrum,
> University of Basel | Klingelbergstrasse 50/70 | CH-4056 Basel
>     Phone: [phone] | Email: [hidden email]<mailto:
> [hidden email]> | www.biozentrum.unibas.ch/imcf<
> http://www.biozentrum.unibas.ch/imcf> | www.microscopynetwork.unibas.ch<
> http://www.microscopynetwork.unibas.ch>
>
>
>

[Steffen Dietzel]

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Oliver,

Are you sure this is *behind* the front lens? have you checked the side
view in a stereo microscope? I had similar crystals on the outside of a
comparable objective when somebody did not suck of the PBS after
imaging. They solved after 15-30 min when I put a Petri dish with
distilled water under the objective and dipped it in. But if that were
your problem I would expect that you should have had at least a little
improvement with 70% Ethanol. You might want to try anyway. If the
sealing is broken you should expect the water to leak into the objective
too, at least over night. Nothing much left to loose in your case, if it
is the sealing.

Steffen


Am 28.05.2018 um 16:38 schrieb Oliver Biehlmaier:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear fellow microscopists,
>
> We have very strange dirt / crystals -image on: https://imgur.com/a/PjOnsSC - on a Zeiss dipping objective which we are using on our 2ph-tomography system.
>
> We tried to clean it with Ethanol 70%, Ethanol 100%, Chloroform, Ether/Ethanol 30%/70% with zero improvement of the situation.
> Thus we guess that the dirt is inside the objective, very much likely salt crystals originating from the buffer solution that the objective is dipping in.
> Have you ever seen something like this? Is it possible that the sealing of the objective is broken after approx. 3 years of dipping and as a consequence these crystals could build up on the rear side of the front lens?
>
> All the best,
> Oliver
>
>
>
> Oliver Biehlmaier, PhD | Head of Imaging Core Facility  | Biozentrum, University of Basel | Klingelbergstrasse 50/70 | CH-4056 Basel
> Phone: [phone] | Email: [hidden email]<mailto:[hidden email]> | www.biozentrum.unibas.ch/imcf<http://www.biozentrum.unibas.ch/imcf> | www.microscopynetwork.unibas.ch<http://www.microscopynetwork.unibas.ch>
>

--
------------------------------------------------------------
Steffen Dietzel, PD Dr. rer. nat
Ludwig-Maximilians-Universität München
Biomedical Center (BMC)
Head of the Core Facility Bioimaging

Großhaderner Straße 9
D-82152 Planegg-Martinsried
Germany

http://www.bioimaging.bmc.med.uni-muenchen.de

[Ferhan A]

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Uninterrupted light reflections at 11 o'clock and 2 o'clock suggest that
the crystals are most probably below the surface (at the back of the lens).
The lens seal seems compromised. Following options may be considered:
1) Preferred way: The front lens needs to be
removed / cleaned / reinserted / resealed (preferably by a professional
service)
2) Quick and dirty way: Via compromised seal, ultrapure water is allowed to
enter/seep into the objective (under vacuum, multiple water changes to
dilute out the salts), then at 37C incubator (+ vacuum, + CaCl2 or silica
as desiccator) ultrapure
water should be evaporated/sucked out completely. (this method will not
prevent re-occurrence)
3) The easiest way: A new objective should be bought and the old one should
be
used as a decorative paper weight (I suggest to try the second option
before this).
Ferhan

On Mon, May 28, 2018 at 4:38 PM, Oliver Biehlmaier <
[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear fellow microscopists,
>
> We have very strange dirt / crystals -image on:
> https://imgur.com/a/PjOnsSC - on a Zeiss dipping objective which we are
> using on our 2ph-tomography system.
>
> We tried to clean it with Ethanol 70%, Ethanol 100%, Chloroform,
> Ether/Ethanol 30%/70% with zero improvement of the situation.
> Thus we guess that the dirt is inside the objective, very much likely salt
> crystals originating from the buffer solution that the objective is dipping
> in.
> Have you ever seen something like this? Is it possible that the sealing of
> the objective is broken after approx. 3 years of dipping and as a
> consequence these crystals could build up on the rear side of the front
> lens?
>
> All the best,
> Oliver
>
>
>
> Oliver Biehlmaier, PhD | Head of Imaging Core Facility  | Biozentrum,
> University of Basel | Klingelbergstrasse 50/70 | CH-4056 Basel
> Phone: [phone] | Email: [hidden email]<mailto:
> [hidden email]> | www.biozentrum.unibas.ch/imcf<
> http://www.biozentrum.unibas.ch/imcf> | www.microscopynetwork.unibas.ch<
> http://www.microscopynetwork.unibas.ch>
>

[Alex Asanov]

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Hi Oliver,

We have seen similar crystals inside our 63X/1.0 water immersion lens,
likely buffer salts. In our case, the sealing was broken. We tried, but
didn't succeed to fix.

Best regards,
Alexander N. Asanov, Ph.D.
President,  TIRF Labs
[hidden email]
www.tirf-labs.com   www.TIRFmicroscopy.com


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Oliver Biehlmaier
Sent: Monday, May 28, 2018 10:39 AM
To: [hidden email]
Subject: Dirty 20x dipping objective on 2-photon system

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Dear fellow microscopists,

We have very strange dirt / crystals -image on: https://imgur.com/a/PjOnsSC
- on a Zeiss dipping objective which we are using on our 2ph-tomography
system.

We tried to clean it with Ethanol 70%, Ethanol 100%, Chloroform,
Ether/Ethanol 30%/70% with zero improvement of the situation.
Thus we guess that the dirt is inside the objective, very much likely salt
crystals originating from the buffer solution that the objective is dipping
in.
Have you ever seen something like this? Is it possible that the sealing of
the objective is broken after approx. 3 years of dipping and as a
consequence these crystals could build up on the rear side of the front
lens?

All the best,
Oliver



Oliver Biehlmaier, PhD | Head of Imaging Core Facility  | Biozentrum,
University of Basel | Klingelbergstrasse 50/70 | CH-4056 Basel
Phone: [phone] | Email:
[hidden email]<mailto:[hidden email]> |
www.biozentrum.unibas.ch/imcf<http://www.biozentrum.unibas.ch/imcf> |
www.microscopynetwork.unibas.ch<http://www.microscopynetwork.unibas.ch>


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