Re: Protective window for a confocal microscope?

Posted by Zdenek Svindrych-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Protective-window-for-a-confocal-microscope-tp7591445p7591462.html

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Hi Tomasz,

back to your question. The filter holder is there to hold a filter. Any
decent glass or fused silica AR window should do the trick.
As far as I remember, the hole is quite big, around 30 mm diameter (bigger
than the standard 25 mm filters), which may limit your options.

Most filters and windows have specified surface flatness (reflected wave
error), but it's actually transmitted wave error (TWE) that matters here.
Usually, windows with single digit flatness (in lambda or fringe units)
will have TWE << 1/5 lambda, and should be OK.

Thicker substrate (2 - 3 mm) usually means precision optics, and is a safe
bet. Hard coating is a must, if you want to wipe oil off of it regularly.
All reputable brands use hard coatings these days, 1% loss (per pass) is
OK.

Sapphire is hard to work with and comes with a price premium. Fused silica,
e.g.
https://www.edmundoptics.com/p/30mm-dia-vis-nir-coated-1lambda-fused-silica-window/10158/
(no commercial interest) looks like a good option, but BK-7 will work well,
too.

If you happen to have a piece of AR-coated glass at hand, just put it there
and look at some beads. If you're not able to tell the difference, you're
good to go. You can even try a glass coverslip, the 4% loss (the excitation
loss is not an issue, you can always crank up the laser) will be hardly
noticeable. Coverslips are not polished, so the TWE is generally bad, but
with some luck you can find a good one in the stack...

Best, zdenek


On Tue, Oct 27, 2020 at 1:18 PM Tomek Węgierski <[hidden email]>
wrote:

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> Dear All,
>
> Thank you very much for so many responses, although the discussion went a
> bit into a direction that I did not anticipate ??
> But yes, it is all about preventing oil from getting into the microscope.
> Our 40x/1.3 Fluar lens takes a lot of oil and I frequently have to clean
> its bottom side from oil. In contrast, I never saw any oil on the back of
> 63x/1.4 even though it is used as frequently as the 40x.
>
> I have to admit I have been skeptical about hair elastics, because when
> they saturate with oil they will just let it go. IMHO, it is replacing "oil
> cleaning" approach with "oil buffering" approach. The "cleaning" approach
> has worked for us for many years. It failed when an unexperienced user (who
> apparently did not take seriously what I was talking during the training)
> made two mistakes: did not clean the lens and let it stay in the optic path
> after the work instead of changing for 10x. In addition, it was a holiday
> time, the system was not used for the next few days, giving the oil plenty
> of time. So, it was a coincidence of several factors, but it can happen
> again.
>
> I think, hearing so many positive opinions about hair elastics, I will
> give them the 2nd chance in our facility. Aqua-Stop in my opinion is
> overpriced, and we do not currently do any experiments under perfusion to
> justify such purchase (this may change in the future). However, I would not
> like to give up on the protective window strategy. I think placing a glass
> element in the infinity space is not unusual. It would function as the last
> stronghold against the oil. And it would be considerably cheaper than
> Aqua-Stop. I would of course test PSF. If PSF is not affected, I guess the
> protective window is OK with imaging. So, if you still have some advice
> which one would be most suitable, I would appreciate it.
>
> Best,
> Tomasz
>
> Tomasz Wegierski, PhD
> International Institute of Molecular and Cell Biology
> Trojdena 4, 02-109 Warsaw, POLAND
> tel: +48-22 597 0763
> fax: +48 22 597 0715
> http://www.iimcb.gov.pl/
> ________________________________
> From: Confocal Microscopy List <[hidden email]> on
> behalf of Frohlich, Victoria <[hidden email]>
> Sent: Tuesday, October 27, 2020 4:38 PM
> To: [hidden email] <[hidden email]>
> Subject: Re: Protective window for a confocal microscope?
>
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>
> Just my 2 cents.
> Scrunchies work well but are not fool-proof.  Cotton not polyester
> material are more absorbent.
> In addition to scrunchies I suggest wrapping the lens barrel near the nose
> of the objective with plumbers tape to prevent oil from seeping into the
> barrel.  It can be done so that the lens retraction still functions.  Then
> once the lens is on the nosepiece of the scope the plumbers tape can be
> wrapped there as well.
>
> I've also used thin rubber sheets or parafilm sheets, as well as filter
> paper (thanks Mike S.!), to draw oil away from the lens and nosepiece.
>
> Best method, however, is to teach users proper oil application and ensure
> they clean up after themselves.
>
> Regards,
> Vickie
>
> Victoria Centonze Frohlich, PhD
> Director, Light Microscopy Shared Resource
> Cell & Tissue Imaging Center
> St. Jude Children’s Research Hospital
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> MS 312
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> “Images were acquired at the Cell & Tissue Imaging Center which is
> supported by SJCRH and NCI P30 CA021765.”
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of Michael Stanley
> Sent: Tuesday, October 27, 2020 8:19 AM
> To: [hidden email]
> Subject: Re: Protective window for a confocal microscope?
>
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> in the 'old days' in the core we used to cut filter paper and then bore a
> hole with a cork-bore slightly smaller than the lens.  slide 2-3 (or as
> many as will fit) layers of the filter paper over the objective.  this will
> not stop a "flood" of oil (which sounds very possible for some) but it will
> hold much more than you would think.  and, the most important feature of
> this little trick is that the oil will show very quickly on the paper and
> hopefully will trigger a response.!
>
> some of the darker filter papers will show the oil better, but this will
> vary by manufacturer.
>
> no commercial interest here, just trying to protect all the optics in the
> system.
>
> michael
>
> c. michael stanley, phd
> senior applications scientist
> chroma technology corp.
> 10 imtec lane
> bellows falls,  vt    05101
> [hidden email]
> [hidden email]
>
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of Gary Laevsky
> Sent: Tuesday, October 27, 2020 10:19 AM
> To: [hidden email]
> Subject: Re: Protective window for a confocal microscope?
>
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>
> The hair ties work great with objectives that have enough of a neck for
> the tie to sit on, next to the barrel.
>
> Our 40X objective (most frequently used) has no/very small neck, so the
> hair ties don't work.
>
> I fairly regularly (bi-weekly) have to take the objective off, store it in
> it's case upside down, and put it in a 37C oven.  Ridiculous, in a bad way,
> how much oil comes out.  The 40X also doesn't have "lip" that acts as a bit
> of a reservoir (unlike the 60 and 100).
>
> On one of my heavily used systems, I have to go so far as to dismantle the
> top (stage, objective turret, and fluorescent turret) to get to the piece
> of glass that prevents leakage into the body (amazingly awesome feature of
> the stand).  Takes me 15 minutes.  Although I shouldn't have to do that ...
>
> On Tue, Oct 27, 2020 at 9:13 AM Cammer, Michael <
> [hidden email]> wrote:
>
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> > Aquastop works well for big spills when installed correctly and has
> > proven effective on some of our Zeiss scopes.  But we did have a user
> > who spilled over or around it on our Zeiss 710, so it is not foolproof.
> >
> >
> > Hair elastics are great for the oil.
> >
> >
> > Michael Cammer, Sr Research Scientist, DART Microscopy Laboratory
> >
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> > NY
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> > ________________________________
> > From: Confocal Microscopy List <[hidden email]> on
> > behalf of Doug Richardson <[hidden email]>
> > Sent: Tuesday, October 27, 2020 7:41:45 AM
> > To: [hidden email]
> > Subject: Re: Protective window for a confocal microscope?
> >
> > [EXTERNAL]
> >
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> > *****
> >
> > I would highly recommend the aquastop system from Zeiss. It is
> > overpriced for some pieces of plastic, but it forms a very tight seal
> > and pays for itself many times over. It is far superior to hair
> > elastics/glove fingers/condoms which we used in the past.
> >
> > Doug
> >
> > Get Outlook for Android
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> > ________________________________
> > From: Confocal Microscopy List <[hidden email]> on
> > behalf of Tomek Węgierski <[hidden email]>
> > Sent: Tuesday, October 27, 2020 7:12:10 AM
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> > Subject: Protective window for a confocal microscope?
> >
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> >
> > Dear All,
> > I am considering a purchase of a protective window for our Zeiss
> > LSM800 and I would like to hear your advice whether this is a good
> > idea, and if yes, which one to buy.
> > The reason is our LSM800 is on an inverted stand and we already got a
> > leakage of oil once which went down onto Optovar. And this was despite
> > my regular checks of objectives (both sides) for leaking oil and
> > extensive education of users on how important it is not to use too
> > much oil and to clean lenses, especially on inverted stands.
> > Zeiss Observer has a dummy slider with an opening for 32 mm filters. I
> > hope, a protective window there should stop the oil, if the leakage
> > happens again.
> > 1) do you think placing a protective window in a confocal microscope
> > is a good idea or not really, because the imaging will be affected?
> > This would be in infinity space, so I guess it should not be
> > detrimental....
> > 2) which characteristics of protective window I should take into
> > consideration (substrate, thickness, coating, flatness, surface
> > quality, parallelism)? Currently I am inclined into protective glasses
> > by Edmund Optics such as 1-mm thick MgF2-coated Sapphire window, which
> > has high transmission, or 2-mm thick MgF2-coated B270 window, which
> > has a bit worse transmission but better surface parameters (and is
> > much cheaper).
> > best regards,
> > Tomasz
> >
> >
> > Tomasz Wegierski, PhD
> > International Institute of Molecular and Cell Biology Trojdena 4,
> > 02-109 Warsaw, POLAND
> > tel: +48-22 597 0763
> > fax: +48 22 597 0715
> >
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> >
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> Best,
>
> Gary Laevsky, Ph.D.
> Director, Confocal Imaging Facility
> Nikon Center of Excellence
> Co-Founder, North Atlantic Microscopy Society (NAMS)
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--
--
Zdenek Svindrych, Ph.D.
Research Scientist - Microscopy Imaging Specialist
Department of Biochemistry and Cell Biology
Geisel School of Medicine at Dartmouth