405 nm laser problems with Zeiss 700

[Cammer, Michael-2]

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We have (or manage) a few Zeiss 700 systems on both upright and inverted microscopes.


We are having persistent problems with the 405 nm laser on two of these scopes.  Actually, we're not sure if the problem is the lasers or other components in the system.  (For instance, we know the alignment of the 405 nm light path on our Zeiss 880 is not stable over weeks, but this is easy to address with manual alignment in the Maintain tab.)


The problem is that sometimes when we turn on the system, it works fine.  Other times it fails and we cannot see anything with the 405 line (and it is not visible at the sample).  Unlike most other problems which are specific to scope users and when staff try, everything works fine, this happens to us too occasionally.  However, Zeiss engineers have not been able to reproduce the problem when they visit, so having it fixed is a problem.


Curious if anyone else has encountered this and, if so, how it was successfully addressed.


Best regards-


Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory

NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY  10016

[hidden email]<mailto:[hidden email]>  http://nyulmc.org/micros  http://microscopynotes.com/

Voice direct only, no text or messages:  1-914-309-3270 and 1-646-501-0567


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[George McNamara]

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Hi Michael,

consider adding BioLegend's Zombie NIR or other far red or NIR DNA
counterstain (To-Pro-5, DRAQ5, other Zombie's, etc possible alternatives).

I also note this is 2020, not 1984: users should only light up the cell
features they really need (and with fluorophores that their favored
microscopes work well with). DAPI, Hoechst (or Zombie, etc) reagents are
pennies per slide or imaging dish, but may note really be needed ---
confocal acquisition time is probably more expensive cost (ex. $27/hr, 2
min of DAPI time is about $1). Antibodies - either flow cytometry style
direct labeled (which even 7-series era scopes should be able to image),
or fluorescent secondary antibodies (see AF+ link below: reported 3x
brighter than the stuff TF/MPE have been selling since 1999; TF of
course just one of many vendors) available in lots of colors, so could
light ip nuclear pores, lamins, histone(s), etc instead.

***

More thoughts for listserv readers on antibodies, or any other reagent
in refrigerator from before pandemic (and refrigerator might have been
the vendor's, not just you or colleagues): Reagents go bad. Would you
eat an egg put in a refrigerator pre-covid-19 pandemic? (or Halloween
2019: Trick or Treat?). I think I've posted these quotes in the past:

Douglas MacArthur: *Old**soldiers**never**die*--they just fade away.
(apparently quoting an 'old army ballad' - URL at bottom).

GM: Old antibodies die, please throw them away.


Happy 2020,

George

p.s. not the first confocal microscope problem a vendor has been unable
or unwilling to fix.

p.p.s. AF+ web link mentioned above (no commercial interest)
https://www.thermofisher.com/us/en/home/life-science/antibodies/secondary-antibodies/fluorescent-secondary-antibodies/alexa-fluor-plus-secondary-antibodies.html

MacArthur quoting an old army ballad according to
https://iowaculture.gov/history/education/educator-resources/primary-source-sets/cold-war/old-soldiers-never-die-address-gen


-------- Forwarded Message --------
Subject: 405 nm laser problems with Zeiss 700
Date: Fri, 23 Oct 2020 15:51:49 +0000
From: Cammer, Michael <[hidden email]>
Reply-To: Confocal Microscopy List <[hidden email]>
To: [hidden email]



*****
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.
*****

We have (or manage) a few Zeiss 700 systems on both upright and inverted
microscopes.


We are having persistent problems with the 405 nm laser on two of these
scopes. Actually, we're not sure if the problem is the lasers or other
components in the system. (For instance, we know the alignment of the
405 nm light path on our Zeiss 880 is not stable over weeks, but this is
easy to address with manual alignment in the Maintain tab.)


The problem is that sometimes when we turn on the system, it works fine.
Other times it fails and we cannot see anything with the 405 line (and
it is not visible at the sample). Unlike most other problems which are
specific to scope users and when staff try, everything works fine, this
happens to us too occasionally. However, Zeiss engineers have not been
able to reproduce the problem when they visit, so having it fixed is a
problem.


Curious if anyone else has encountered this and, if so, how it was
successfully addressed.


Best regards-


Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory

NYU Langone Health, 540 First Avenue, SK2 Microscopy Suite, New York, NY
10016

[hidden email]<mailto:[hidden email]>
http://nyulmc.org/micros http://microscopynotes.com/

Voice direct only, no text or messages: 1-914-309-3270 and 1-646-501-0567


------------------------------------------------------------
This email message, including any attachments, is for the sole use of
the intended recipient(s) and may contain information that is
proprietary, confidential, and exempt from disclosure under applicable
law. Any unauthorized review, use, disclosure, or distribution is
prohibited. If you have received this email in error please notify the
sender by return email and delete the original message. Please note, the
recipient should check this email and any attachments for the presence
of viruses. The organization accepts no liability for any damage caused
by any virus transmitted by this email.
=================================