Confocal Microscopy List

Cotton wool for lens cleaning

Classic

List

Threaded

38 messages Options

Stanislav Vitha

Re: Cotton wool for lens cleaning

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I agree,
ether is not good for you. I should have mentioned the obvious - that there
should not be any open flame in the room, and that I do the objective cleaning
in a chemical hood.

Stan Vitha

On Wed, 16 Mar 2011 21:22:59 -0400, Nina Allen <[hidden email]>
wrote:

>Long ago we taught using ether in the fashion described here. Ether is very
flammable. It is also not good for you.

>So even if it works well it is not a recommended method.
>
>Nina Allen
>Professor Emerita
>Department of Plant Biology
>North Carolina State University
>
>Sent from my iPhone
>
>On Mar 16, 2011, at 6:45 PM, Stanislav Vitha <[hidden email]> wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> Hi Aleksandrs,
>>
>> I like to use the method where you clean the lenses without touching them.
>> I learned this from Karl Aufderheide when he was showing this trick to my

LM
>> course students.
>>
>> 1. Don't touch anything (even lens paper) to a lens surface except as a
last
>> resort. Avoid especially commercial facial or bathroom tissue because it
could
>> contain diatom frustules (glass!) as a filler. One pass of a kleenex over a
lens
>> could possibly ruin it!
>> 2. Hold a piece of lens paper or other tissue over a lens. Place a few drops
of
>> ethyl ether on the paper and draw the paper across the lens surface so
that
>> the ether flows rapidly in a circular pattern over the recessed lens surface.
In
>> this way, the ether contacts the lens but the paper does not, because the
>> lens is recessed.
>> 3. Inspect the lens using an inverted ocular as a magnifier. Repeat the
ether
>> wash if necessary.
>> 4. If ether does not remove the dirt, try first distilled water, then
chloroform,
>> then xylene or benzene, in that order. If all else fails, try a 1:1:1 mixture of
>> water, alcohol and chloroform shaken just before use. Follow with an ether
>> wash.
>> 5. For stubborn dirt (e.g., on old student microscopes) use the above
solvents
>> on a clean Q-tip.
>>
>>
>> Because of safety concerns with ether (formation of explosive peroxides), I
>> just get a fresh bottle every 6 months, and dispose of the old one through
our

>> Hazardous Waste program.
>>
>> Stan Vitha
>> Microscopy and Imaging Center
>> Texas A&M University
>>
>> On Tue, 15 Mar 2011 15:03:31 -0400, Aleksandrs Spurmanis, Mr.
>> <[hidden email]> wrote:
>>
>>> Dear list,
>>>
>>> The current practice at our facility is to inspect and clean the objectives

of
>> our scopes periodically (approx. once every 2-3 months for each scope)
using
>> lens paper wrapped around small clean-room swabs. I had noticed,
however,
>> that the field service technicians who run the PMs on our instruments tend
to
>> use 100% cotton wool (which I understand to be essentially the same
material
>> as your basic 100% cotton ball in the pharmacy) and are able to service
our
>> lenses in a much more efficient manner (read: waayyy quicker) than myself
>> using my current methods. In the interests of improving my maintenance
>> efficiency, I've been considering trying this out myself but wanted to check
in
>> with the list to see if anyone can share their experiences, insights or
advice
>> before proceeding. My main concern is that the cotton might contribute to
>> premature wear on the lens coating. As cleaning solvents, I use either
Glass

>> Plus, anhydrous ethanol and/or water.
>>>
>>> Thanks in advance.
>>>
>>> Sincerely,
>>>
>>> Aleksandrs J. Spurmanis
>>> Microscopy Specialist
>>> Imaging Facility
>>> McGill University Life Sciences Complex
>>> Francesco Bellini Building
>>> 3649 Sir William Osler
>>> Suite 137
>>> Montreal, QC
>>> H3G 0B1
>>> tel.: (514)-398-5248
>>> fax: (514)-398-7452
>>> [hidden email]
>>> http://www.mcgill.ca/lifesciencescomplex/core/imaging/

Craig Brideau

Re: Cotton wool for lens cleaning

In reply to this post by Mark Cannell

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I agree with Mark. Acetone will eat any rubbers/sealers/epoxies holding
your lens together. I only use acetone on mirrors out on the optical bench
that are held in place with friction mounts (i.e. no epoxy). For anything
glued or sealed I use high-purity methanol and lens tissues.

Craig

On Thu, Mar 17, 2011 at 7:09 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
> *****
>
> From direct experience, I urge a note of caution with acetone, you may
> degrade the lens mounting glue and risk spreading it over the lens.
>
> Regards Mark
>
>
> On 17/03/2011, at 12:40 PM, Cammer, Michael wrote:
>
> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> I don't think there really is one general solvent. When I began doing
>> biological microscopy about 20 years ago the immersion oils all seemed to be
>> made of the same stuff and easily cleaned with just about any organic
>> solvent stronger than ethanol. This is no longer the case. For instance,
>> the new Nikon oil for TIRF gets thick and is completely impervious to any of
>> the aqueous cleaners. It is resistant to what we practically considered to
>> be the universal solvent of organics, acetone, and also to ethanol. But
>> dehydrated methanol works great. On the other hand, the Zeiss oils, when
>> fresh, clean up fine with their aqueous cleaning solutions and when old and
>> dripped all over the turret and such, with acetone. The old standby in the
>> lab, Cargill Labs type DF, cleans up with any inorganic solvent. Of course,
>> in one lab the gospel was xylene because, well, we scientists tend to be
>> superstitious or traditional. As for ether, one benefit of using it, we
>> were told years ago by someone at Zeiss, is that it evaporates so fast that
>> it reduces the chances of dissolving the glue holding in the front glass of
>> the objective. Is this really a problem? I've never had one of these front
>> lenses come loose. Now I tend to use 1:1 acetone/methanol and cotton swabs
>> and/or lens tissue following in the footsteps of Spectraphysics service who
>> uses this to clean their mirrors and gives us average power of a Watt with
>> 100 fs pulses at 910-920 nm, so I follow by example.
>> -Michael Cammer
>>
>> ------------------------------------------------------------
>> 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.
>> =================================
>>
>

Keith Morris

Re: Cotton wool for lens cleaning

In reply to this post by Stanislav Vitha

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Regarding ether hazards when cleaning the top objective lens, I do only use
a working 10ml bottle of ether [stored in our flammables cabinet], not a 2.5
l winchester - and I open the lid, dip in the cotton bud and re-screw the
lid immediately [I don't pour it onto lens tissues]. The advice is to use
diethyl ether in 'well ventilated areas' [rather than 'fume cupboard' only].
Ether explosion limits are 1.7% to 48% compared to alcohols 3.3% to 24.5%
[although ether does evaporates at a far higher rate]. In our lab should the
whole 10ml evaporate the air concentration would be about 0.00035%, around
5,000 times lower than the explosion limit, and the room is well ventilated.

That said diethyl ether boils at 34.6oC and being denser than air its
creeping, volatile and highly flammable nature is pretty impressive - see
The Ether Trough video at
http://www.angelo.edu/faculty/kboudrea/demos/ether_trough/ether_trough.htm
Hence Stan's important comments on avoiding naked flames in the lab when
ether is about.

I suppose absolute alcohol is also highly flammable, and here we have litres
of that about in the lab. Diethyl ether has additives to reduce explosive
byproducts [unstable peroxides left after evaporation].

Methanol is toxic by inhalation, rather than harmful as for ether and
alcohol, and personally I probably wouldn't use it over ether or alcohol.
But again it's small volumes in the fume cupboard, and we use
methanol/acetic acid a lot as a fixative.

For an interesting discussion of ether used as a recreational drug [it's
effects are similar to alcohol, although you sober up far faster] see:
http://www.druglibrary.org/schaffer/library/studies/cu/cu43.html

Regards

Keith

Advice for labs, schools and colleges:
http://cartwright.chem.ox.ac.uk/hsci/chemicals/ethyl_alcohol.html
http://cartwright.chem.ox.ac.uk/hsci/chemicals/diethyl_ether.html
http://cartwright.chem.ox.ac.uk/hsci/chemicals/methanol.html
http://cartwright.chem.ox.ac.uk/hsci/chemicals/hsci_chemicals_list.html

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford OX3 7BN,
United Kingdom.

Telephone: +44 (0)1865 287568
Email: [hidden email]
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Stanislav Vitha
Sent: 17 March 2011 14:50
To: [hidden email]
Subject: Re: Cotton wool for lens cleaning

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I agree,
ether is not good for you. I should have mentioned the obvious - that there
should not be any open flame in the room, and that I do the objective
cleaning
in a chemical hood.

Stan Vitha

On Wed, 16 Mar 2011 21:22:59 -0400, Nina Allen <[hidden email]>
wrote:

>Long ago we taught using ether in the fashion described here. Ether is
very
flammable. It is also not good for you.

them.
>> I learned this from Karl Aufderheide when he was showing this trick to my

LM
>> course students.
>>
>> 1. Don't touch anything (even lens paper) to a lens surface except as a
last
>> resort. Avoid especially commercial facial or bathroom tissue because it
could
>> contain diatom frustules (glass!) as a filler. One pass of a kleenex over
a
lens
>> could possibly ruin it!
>> 2. Hold a piece of lens paper or other tissue over a lens. Place a few
drops
of
>> ethyl ether on the paper and draw the paper across the lens surface so
that
>> the ether flows rapidly in a circular pattern over the recessed lens
surface.
In
>> this way, the ether contacts the lens but the paper does not, because the

>> lens is recessed.
>> 3. Inspect the lens using an inverted ocular as a magnifier. Repeat the
ether
>> wash if necessary.
>> 4. If ether does not remove the dirt, try first distilled water, then
chloroform,
>> then xylene or benzene, in that order. If all else fails, try a 1:1:1
mixture of
>> water, alcohol and chloroform shaken just before use. Follow with an
ether
>> wash.
>> 5. For stubborn dirt (e.g., on old student microscopes) use the above
solvents
>> on a clean Q-tip.
>>
>>
>> Because of safety concerns with ether (formation of explosive peroxides),
I
>> just get a fresh bottle every 6 months, and dispose of the old one
through
our

objectives
of
>> our scopes periodically (approx. once every 2-3 months for each scope)
using
>> lens paper wrapped around small clean-room swabs. I had noticed,
however,
>> that the field service technicians who run the PMs on our instruments
tend
to
>> use 100% cotton wool (which I understand to be essentially the same
material
>> as your basic 100% cotton ball in the pharmacy) and are able to service
our
>> lenses in a much more efficient manner (read: waayyy quicker) than myself

>> using my current methods. In the interests of improving my maintenance
>> efficiency, I've been considering trying this out myself but wanted to
check
in
>> with the list to see if anyone can share their experiences, insights or
advice
>> before proceeding. My main concern is that the cotton might contribute
to
>> premature wear on the lens coating. As cleaning solvents, I use either
Glass

David Coder-2

Re: Cotton wool for lens cleaning

In reply to this post by Aleksandrs Spurmanis, Mr.

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

"The only thing that really worried me was the ether. There is nothing in
the world more helpless and irresponsible and depraved than a man in the
depths of an ether binge, and I knew we'd get into that rotten stuff pretty
soon."

The perils of organic solvents have been discussed, but I also recall that
alcohols (perhaps only EtOH?) pose a risk for some lens cements. Perhaps
lens manufacturers might comment.

Aqueous solvents seem to fairly benign: Kodak lens cleaner (and lens paper)
has been around for some time. I also remember the recommendation of a
skilled microscope technician who used Windex (another proprietary glass
cleaner) after leaving the bottle open for sufficient time to let the
ammonia evaporate.

(A check of the web turns up the following for an early? forumulation of
Windex at Wikipedia:
"The Sam Wise patent #3,463,735 lists several example formulae, one of which
is 4.0% isopropyl alcohol <http://en.wikipedia.org/wiki/Isopropyl_alcohol> (a
highly volatile solvent) 1% ethylene glycol monobutyl
ether<http://en.wikipedia.org/wiki/2-Butoxyethanol> (a
less volatile solvent), 0.1% sodium lauryl
sulfate<http://en.wikipedia.org/wiki/Sodium_lauryl_sulfate> (a
surfactant), 0.01% tetrasodium
pyrophosphate<http://en.wikipedia.org/wiki/Tetrasodium_pyrophosphate>
(a
water softener), 0.05% of 28%ammonia <http://en.wikipedia.org/wiki/Ammonia>,
1% of a dye solution, and 0.01% perfume.")

And I also agree with general advice of not touching the surface unless you
must. And then only with lens paper or cotton (AKA cotton wool). As for
cotton swabs/buds, I'd be cautious given the adhesive to keep the cotton on
the stick, plastic, or rolled paper rod (as in Q-tips).

*________________________*
David M. Coder, PhD
for Bay bioscience, Co.
www.baybio.co.jp <http://www.baybio.co.jp/english/top.html>
Irvine, CA
Cell phone: 949 233 2641
Skype: dave.coder
Email: [hidden email]

On Fri, Mar 18, 2011 at 10:03 PM, CONFOCALMICROSCOPY automatic digest system
<[hidden email]> wrote:

> Date: Sat, 19 Mar 2011 00:03:49 -0500
> Reply-To: Confocal Microscopy List <[hidden email]>
> Sender: Confocal Microscopy List <[hidden email]>
> From: CONFOCALMICROSCOPY automatic digest system <
> [hidden email]>
> Subject: CONFOCALMICROSCOPY Digest - 17 Mar 2011 to 18 Mar 2011 (#2011-70)
> To: [hidden email]
> Message-ID: <CONFOCALMICROSCOPY%[hidden email]>
> Precedence: list
> List-Help: <http://lists.umn.edu/cgi-bin/wa?LIST=CONFOCALMICROSCOPY>,
> <mailto:[hidden email]?body=INFO%20CONFOCALMICROSCOPY>
> List-Unsubscribe: <mailto:
> [hidden email]>
> List-Subscribe: <mailto:[hidden email]
> >
> List-Owner: <mailto:[hidden email]>
> List-Archive: <http://lists.umn.edu/cgi-bin/wa?LIST=CONFOCALMICROSCOPY>
>
> There is 1 message totalling 255 lines in this issue.
>
> Topics of the day:
>
> 1. Cotton wool for lens cleaning
>
> Date: Fri, 18 Mar 2011 17:06:36 -0000
> From: Keith Morris <[hidden email]>
> Subject: Re: Cotton wool for lens cleaning
> MIME-Version: 1.0
> Content-Type: text/plain; charset="iso-8859-1"
> Content-Transfer-Encoding: quoted-printable
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=3Dconfocalmicroscopy
> *****
>
> Regarding ether hazards when cleaning the top objective lens, I do only =
> use
> a working 10ml bottle of ether [stored in our flammables cabinet], not a =
> 2.5
> l winchester - and I open the lid, dip in the cotton bud and re-screw =
> the
> lid immediately [I don't pour it onto lens tissues]. The advice is to =
> use
> diethyl ether in 'well ventilated areas' [rather than 'fume cupboard' =
> only].
> Ether explosion limits are 1.7% to 48% compared to alcohols 3.3% to =
> 24.5%
> [although ether does evaporates at a far higher rate]. In our lab should =
> the
> whole 10ml evaporate the air concentration would be about 0.00035%, =
> around
> 5,000 times lower than the explosion limit, and the room is well =
> ventilated.
>
>
> That said diethyl ether boils at 34.6oC and being denser than air its
> creeping, volatile and highly flammable nature is pretty impressive - =
> see
> The Ether Trough video at
> http://www.angelo.edu/faculty/kboudrea/demos/ether_trough/ether_trough.ht=
> m
> Hence Stan's important comments on avoiding naked flames in the lab when
> ether is about.
>
> I suppose absolute alcohol is also highly flammable, and here we have =
> litres
> of that about in the lab. Diethyl ether has additives to reduce =
> explosive
> byproducts [unstable peroxides left after evaporation].=20
>
> Methanol is toxic by inhalation, rather than harmful as for ether and
> alcohol, and personally I probably wouldn't use it over ether or =
> alcohol.
> But again it's small volumes in the fume cupboard, and we use
> methanol/acetic acid a lot as a fixative.=20
>
> For an interesting discussion of ether used as a recreational drug [it's
> effects are similar to alcohol, although you sober up far faster] see:
> http://www.druglibrary.org/schaffer/library/studies/cu/cu43.html
>
> Regards
>
> Keith
>
> Advice for labs, schools and colleges:
> http://cartwright.chem.ox.ac.uk/hsci/chemicals/ethyl_alcohol.html
> http://cartwright.chem.ox.ac.uk/hsci/chemicals/diethyl_ether.html
> http://cartwright.chem.ox.ac.uk/hsci/chemicals/methanol.html
> http://cartwright.chem.ox.ac.uk/hsci/chemicals/hsci_chemicals_list.html
>
> -------------------------------------------------------------------------=
> --
> Dr Keith J. Morris,
> Molecular Cytogenetics and Microscopy Core,
> Laboratory 00/069 and 00/070,
> The Wellcome Trust Centre for Human Genetics,
> Roosevelt Drive,
> Oxford=A0 OX3 7BN,
> United Kingdom.
>
> Telephone:=A0 +44 (0)1865 287568
> Email:=A0 [hidden email]
> Web-pages: =
> http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy
>
>
> -----Original Message-----
>
> *

Jeremy Adler-4

Vivatome experience

In reply to this post by Zoltan

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Zeiss and Andor have an aperture correlation microscope that has
optical sectioning capability, based on subtracting a widefield image
from a widefield + infocus image.
The attraction, compared to a point scanning confocal, is speed,
efficient use of photons and low cost.

What experience users have and how do PSFs compare with a point
scanning confocal.

Grant MacGregor

Re: Cotton wool for lens cleaning

In reply to this post by Craig Brideau

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I concur with Craig and Mark regarding use of ether and any other solvent of lens cements. Many years ago I watched the front lens of a 1.4NA condenser fall out following too much elbow grease with ether-soaked cotton buds / Q-tips. Ether no more.

Several years ago someone dumped a plate full of tissue culture media down the nosepiece of a shared facility's vert 200m I was in charge of overseeing. Prior to fitting the aqua stop, I watched with great interest to see what the extremely senior, German accented and fussy Zeiss field service engineer would use to clean the optics. Would it be Opti L ? Would it be pure ether ? Surprise surprise, he whipped out a large spray bottle of Sparkle. This is a surfactant-laden ammonia free solution one can purchase online or at an Ace Hardware (if in the US). The regular Sparkle is purple. A while ago I suspect the makers of Sparkle found out that folks were using it for high-end optics as they started packaging it in smaller (c. 100ml) bottles and leaving out the purple dye. I don't know if they charge more for this, but I would assume so. I discovered if you leave the bottle of purple Sparkle in the window, the purple dye (presumably) oxidises with a few weeks, becoming colorless.

For what it is worth, our general rule of thumb for cleaning optics is as follows.

Try in order the following solvents - 1) distilled water; 2) Sparkle. If we have to use Sparkle, we follow up with distilled water, which can be simply applied by breathing on the optic.
Clean using only pure cotton or silk and not lens paper. I was taught that lens paper should be used only for storage of optics. Back in the day, microscope field engineers often wore silk ties that had a bunch of stains on the back of them.
Clean in a circular manner in a single direction, taking care not to drag any dirt back over the cleaned area.

Grant MacGregor
Developmental and Cell Biology,
UC Irvine

On Mar 17, 2011, at 12:04 PM, Craig Brideau wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I agree with Mark. Acetone will eat any rubbers/sealers/epoxies holding
> your lens together. I only use acetone on mirrors out on the optical bench
> that are held in place with friction mounts (i.e. no epoxy). For anything
> glued or sealed I use high-purity methanol and lens tissues.
>
> Craig
>
>
> On Thu, Mar 17, 2011 at 7:09 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
>> *****
>>
>> From direct experience, I urge a note of caution with acetone, you may
>> degrade the lens mounting glue and risk spreading it over the lens.
>>
>> Regards Mark
>>
>>
>> On 17/03/2011, at 12:40 PM, Cammer, Michael wrote:
>>
>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>> *****
>>>
>>> I don't think there really is one general solvent. When I began doing
>>> biological microscopy about 20 years ago the immersion oils all seemed to be
>>> made of the same stuff and easily cleaned with just about any organic
>>> solvent stronger than ethanol. This is no longer the case. For instance,
>>> the new Nikon oil for TIRF gets thick and is completely impervious to any of
>>> the aqueous cleaners. It is resistant to what we practically considered to
>>> be the universal solvent of organics, acetone, and also to ethanol. But
>>> dehydrated methanol works great. On the other hand, the Zeiss oils, when
>>> fresh, clean up fine with their aqueous cleaning solutions and when old and
>>> dripped all over the turret and such, with acetone. The old standby in the
>>> lab, Cargill Labs type DF, cleans up with any inorganic solvent. Of course,
>>> in one lab the gospel was xylene because, well, we scientists tend to be
>>> superstitious or traditional. As for ether, one benefit of using it, we
>>> were told years ago by someone at Zeiss, is that it evaporates so fast that
>>> it reduces the chances of dissolving the glue holding in the front glass of
>>> the objective. Is this really a problem? I've never had one of these front
>>> lenses come loose. Now I tend to use 1:1 acetone/methanol and cotton swabs
>>> and/or lens tissue following in the footsteps of Spectraphysics service who
>>> uses this to clean their mirrors and gives us average power of a Watt with
>>> 100 fs pulses at 910-920 nm, so I follow by example.
>>> -Michael Cammer
>>>
>>> ------------------------------------------------------------
>>> 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.
>>> =================================
>>>
>>
>

Dan Focht

Re: Cotton wool for lens cleaning

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

On the subject of not using lens paper, I want to offer the suggestion of using Berkshire Lensx90.

I agree that most so called lens paper is unsuited for microscope or any other type of lens.
I can't believe they sell that hard, dense, stiff stuff as lens paper.
However, in a former life when I serviced high end microscopes, I found that LensX90 can't be beat for softness, absorbency and freedom from detrimental paper additives that can scratch microscope objectives.
It comes in a 4 X 6 inch sheet.
I would cut the paper down to 2x6" and wrap one piece at a time on the end of a 1/16" wooden stick such that there was about 10mm of wrapped soft paper past the end of the stick.
It could then be used as an applicator or wipe.
You can then pull the paper off of the stick and apply a new paper every time it was used.
I still use it today.

http://www.berkshire.com/lensx90.shtml

I have no financial interest in the company, I just hope they don't fall victim to todays economy!

Dan

On Mar 21, 2011, at 3:19 PM, Grant MacGregor wrote:

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I concur with Craig and Mark regarding use of ether and any other solvent of lens cements. Many years ago I watched the front lens of a 1.4NA condenser fall out following too much elbow grease with ether-soaked cotton buds / Q-tips. Ether no more.

Several years ago someone dumped a plate full of tissue culture media down the nosepiece of a shared facility's vert 200m I was in charge of overseeing. Prior to fitting the aqua stop, I watched with great interest to see what the extremely senior, German accented and fussy Zeiss field service engineer would use to clean the optics. Would it be Opti L ? Would it be pure ether ? Surprise surprise, he whipped out a large spray bottle of Sparkle. This is a surfactant-laden ammonia free solution one can purchase online or at an Ace Hardware (if in the US). The regular Sparkle is purple. A while ago I suspect the makers of Sparkle found out that folks were using it for high-end optics as they started packaging it in smaller (c. 100ml) bottles and leaving out the purple dye. I don't know if they charge more for this, but I would assume so. I discovered if you leave the bottle of purple Sparkle in the window, the purple dye (presumably) oxidises with a few weeks, becoming colorless.

For what it is worth, our general rule of thumb for cleaning optics is as follows.

Try in order the following solvents - 1) distilled water; 2) Sparkle. If we have to use Sparkle, we follow up with distilled water, which can be simply applied by breathing on the optic.
Clean using only pure cotton or silk and not lens paper. I was taught that lens paper should be used only for storage of optics. Back in the day, microscope field engineers often wore silk ties that had a bunch of stains on the back of them.
Clean in a circular manner in a single direction, taking care not to drag any dirt back over the cleaned area.

Grant MacGregor
Developmental and Cell Biology,
UC Irvine

On Mar 17, 2011, at 12:04 PM, Craig Brideau wrote:

Dan Focht
Bioptechs, Inc.
3560 Beck Rd.
Butler, PA 16002
www.bioptechs.com
P: (724)282-7145
F: (724)282-0745
[hidden email]

Glen MacDonald-2

Re: Cotton wool for lens cleaning

In reply to this post by Grant MacGregor

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Not all Ace hardware stores carry Sparkle, only some Tru-Valu hardware stores carry it, no one in Canada has it. You can buy from their website, including gallon refills. I repackage into plastic dropper bottles for the lab bench.
http://www.glasscleaner.com/

The clear and purple seem to behave a little differently, but maybe that is just placebo effect. The optical cleaner seems to do a better job with Cargille Type B oil but more prone to leave spots than does the purple stuff. I usually follow up the clear cleaner with distilled water. The purple cleaner seems better for some of the other materials I've had to take off the lenses.

We banned chloroform (preferred by a former Zeiss service engineer) after someone applied it to an inexpensive condenser on a Nikon TMD inverted scope. It smeared the surface of a plastic lens.
Glen

Glen MacDonald
Core for Communication Research
Virginia Merrill Bloedel Hearing Research Center
Box 357923
University of Washington
Seattle, WA 98195-7923 USA
(206) 616-4156
[hidden email]

On Mar 21, 2011, at 12:19 PM, Grant MacGregor wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I concur with Craig and Mark regarding use of ether and any other solvent of lens cements. Many years ago I watched the front lens of a 1.4NA condenser fall out following too much elbow grease with ether-soaked cotton buds / Q-tips. Ether no more.
>
> Several years ago someone dumped a plate full of tissue culture media down the nosepiece of a shared facility's vert 200m I was in charge of overseeing. Prior to fitting the aqua stop, I watched with great interest to see what the extremely senior, German accented and fussy Zeiss field service engineer would use to clean the optics. Would it be Opti L ? Would it be pure ether ? Surprise surprise, he whipped out a large spray bottle of Sparkle. This is a surfactant-laden ammonia free solution one can purchase online or at an Ace Hardware (if in the US). The regular Sparkle is purple. A while ago I suspect the makers of Sparkle found out that folks were using it for high-end optics as they started packaging it in smaller (c. 100ml) bottles and leaving out the purple dye. I don't know if they charge more for this, but I would assume so. I discovered if you leave the bottle of purple Sparkle in the window, the purple dye (presumably) oxidises with a few weeks, becoming colorless.
>
> For what it is worth, our general rule of thumb for cleaning optics is as follows.
>
> Try in order the following solvents - 1) distilled water; 2) Sparkle. If we have to use Sparkle, we follow up with distilled water, which can be simply applied by breathing on the optic.
> Clean using only pure cotton or silk and not lens paper. I was taught that lens paper should be used only for storage of optics. Back in the day, microscope field engineers often wore silk ties that had a bunch of stains on the back of them.
> Clean in a circular manner in a single direction, taking care not to drag any dirt back over the cleaned area.
>
> Grant MacGregor
> Developmental and Cell Biology,
> UC Irvine
>
>
>
> On Mar 17, 2011, at 12:04 PM, Craig Brideau wrote:
>
>> *****
>> To join, leave or search the confocal microscopy listserv, go to:
>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>> *****
>>
>> I agree with Mark. Acetone will eat any rubbers/sealers/epoxies holding
>> your lens together. I only use acetone on mirrors out on the optical bench
>> that are held in place with friction mounts (i.e. no epoxy). For anything
>> glued or sealed I use high-purity methanol and lens tissues.
>>
>> Craig
>>
>>
>> On Thu, Mar 17, 2011 at 7:09 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
>>> *****
>>>
>>> From direct experience, I urge a note of caution with acetone, you may
>>> degrade the lens mounting glue and risk spreading it over the lens.
>>>
>>> Regards Mark
>>>
>>>
>>> On 17/03/2011, at 12:40 PM, Cammer, Michael wrote:
>>>
>>> *****
>>>> To join, leave or search the confocal microscopy listserv, go to:
>>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>>> *****
>>>>
>>>> I don't think there really is one general solvent. When I began doing
>>>> biological microscopy about 20 years ago the immersion oils all seemed to be
>>>> made of the same stuff and easily cleaned with just about any organic
>>>> solvent stronger than ethanol. This is no longer the case. For instance,
>>>> the new Nikon oil for TIRF gets thick and is completely impervious to any of
>>>> the aqueous cleaners. It is resistant to what we practically considered to
>>>> be the universal solvent of organics, acetone, and also to ethanol. But
>>>> dehydrated methanol works great. On the other hand, the Zeiss oils, when
>>>> fresh, clean up fine with their aqueous cleaning solutions and when old and
>>>> dripped all over the turret and such, with acetone. The old standby in the
>>>> lab, Cargill Labs type DF, cleans up with any inorganic solvent. Of course,
>>>> in one lab the gospel was xylene because, well, we scientists tend to be
>>>> superstitious or traditional. As for ether, one benefit of using it, we
>>>> were told years ago by someone at Zeiss, is that it evaporates so fast that
>>>> it reduces the chances of dissolving the glue holding in the front glass of
>>>> the objective. Is this really a problem? I've never had one of these front
>>>> lenses come loose. Now I tend to use 1:1 acetone/methanol and cotton swabs
>>>> and/or lens tissue following in the footsteps of Spectraphysics service who
>>>> uses this to clean their mirrors and gives us average power of a Watt with
>>>> 100 fs pulses at 910-920 nm, so I follow by example.
>>>> -Michael Cammer
>>>>
>>>> ------------------------------------------------------------
>>>> 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.
>>>> =================================
>>>>
>>>
>>

kspencer007

Re: Cotton wool for lens cleaning

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I'll join in...
Has anyone worked with methyl ethyl ketone? I use this only as a weapon of last resort, for really nasty, can't-otherwise-clean unknown gunk on core scope objectives. I use VWR cotton swabs, with a minimal amount of MEK, staying away from any glue/plastic/non-glass surface. For routine cleaning (once a month), I use isopropanol on lens tissue. Is this bad?
Kathy
The Scripps Research Institute

JOEL B. SHEFFIELD

Re: Cotton wool for lens cleaning

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

This has been a fascinating discussion. It occurs to me, though, that we
may be trying to find the perfect cleaning method that will apply to many
different kinds of contamination. I can think of several that seem to me to
be mutually exclusive.

1. Water soluble --or once soluble. I am thinking of dried salts from
medium, from bathing solutions, tears of operators, etc.

2. Oil, deliberate on oil immersion lenses, or accidental, from
inadvertently dipping a dry lens where it shouldn't go.

3. Dried nail polish or other mounting media, inappropriately applied, or
from slides viewed by impatient students (!!!).

At any rate, each of these probably needs a different approach. Organic
solvents won't do much for type 2, and "Sparkle" won't make type 3 sparkle.

Joel

On Mon, Mar 21, 2011 at 6:19 PM, Kathryn Spencer <[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I'll join in...
> Has anyone worked with methyl ethyl ketone? I use this only as a
> weapon of last resort, for really nasty, can't-otherwise-clean unknown gunk
> on core scope objectives. I use VWR cotton swabs, with a minimal amount of
> MEK, staying away from any glue/plastic/non-glass surface. For routine
> cleaning (once a month), I use isopropanol on lens tissue. Is this bad?
> Kathy
> The Scripps Research Institute
>

--

Joel B. Sheffield, Ph.D
Department of Biology
Temple University
Philadelphia, PA 19122
Voice: 215 204 8839
e-mail: [hidden email]
URL: http://astro.temple.edu/~jbs

Lutz Schaefer

Re: Vivatome experience

In reply to this post by Jeremy Adler-4

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Jeremy,

just a slight correction to your description. The optical sectioning
principle is comparable to that of incoherent structured illumination
microscopy. Instead of taking >= 3 images of different grating shifts as one
particular way to solve the linear system of equations which solution
finally leads to a sectioned image, the demodulation in the aperture
correlation microscope is actually done on the camera chip - namely by
integration of the structured intensities. The structured illumination comes
from a spinning disk through which at first the sample is illuminated. The
emitted, structured light from the sample passes again the same disk, but
this time the transmitted and reflected light from the disk is recorded
separately, either with two different cameras or on the same chip on half of
either side.

Now to the correction of your statement, it turns out that both so collected
images have a widefield and a sectioned component. One of the images is the
sum and the other the difference of both components. This enables for a
simple subtraction to cancel out the widefield (to get the sectioned) - or a
simple addition to cancel out the sectioned parts (to get the widefield). To
ensure correct balance of the two slightly different paths of light, there
will have to be a calibrated correction factor involved.

The PSF I would expect to be similar to that of other incoherent structured
illumination microscopy systems (e.g. Zeiss ApoTome).

Hope this was a bit of help to clarify the method
Lutz

__________________________________
L u t z S c h a e f e r
Sen. Scientist
Mathematical modeling / Image processing
Advanced Imaging Methodology Consultation
16-715 Doon Village Rd.
Kitchener, ON, N2P 2A2, Canada
Phone/Fax: +1 519 894 8870
Email: [hidden email]
Website: http://home.golden.net/~lschafer/
___________________________________
--------------------------------------------------
From: "Jeremy Adler" <[hidden email]>
Sent: Monday, March 21, 2011 10:03
To: <[hidden email]>
Subject: Vivatome experience

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Zeiss and Andor have an aperture correlation microscope that has optical
> sectioning capability, based on subtracting a widefield image from a
> widefield + infocus image.
> The attraction, compared to a point scanning confocal, is speed,
> efficient use of photons and low cost.
>
> What experience users have and how do PSFs compare with a point scanning
> confocal.

Daniel James White

Re: Vivatome experience

In reply to this post by Jeremy Adler-4

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi Jeremy,

On Mar 22, 2011, at 6:06 AM, CONFOCALMICROSCOPY automatic digest system wrote:

>
> Date: Mon, 21 Mar 2011 15:03:42 +0100
> From: Jeremy Adler <[hidden email]>
> Subject: Vivatome experience
>
> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Zeiss and Andor have an aperture correlation microscope that has
> optical sectioning capability, based on subtracting a widefield image
> from a widefield + infocus image.
> The attraction, compared to a point scanning confocal, is speed,
> efficient use of photons and low cost.
>
> What experience users have and how do PSFs compare with a point
> scanning confocal.

We have tested in house the Andor DSD and have seen the Zeiss Vivatome in the flash also....
so what to expect from this technology.

Like any other mechanical or mixed mechanical/computational blur removal gadget
photons are removed from the signal... hopefully mostly out of focus ones, leaving in focus signal in the result image.
Thus, you must expect the signal:noise of the result image to bee significantly less than the
2 input images used to create the result image.
Similarly to Apotome, single/multi point scanning confocal, signal is effectively thrown away
in order to get at better contrast especially in the z direction.

So... once your sample is pretty to very bright / highly stained
you can expect good results at a frame rate somewhere between
widefield and apotome, and definitely faster than single point scanners...
...possibly similar frame rate to a Nipkow spinning disk, eg Yokogawa.
But of course its comapring apples and oranges,
and as always it very much depends on the sample.

Advantages are: low cost, low complexity, flexibility, speed. (possibly an unverified theoretical bonus of pulsed illumination at each point of the sample, as seen in spinning disk confocal)
Disadvantages are: weak samples will give very noisy results.

It cant replace a point scanning confocal, or a good spinning disk confocal,
but it is certainly a complementary technology to the existing methods, and will find its niche.

PSF... well, it depends on a few things in theory.... Andor allows you to vary the amount of subtraction of the blurry image... and that will affect the PSF.
But I guess it could possibly be similar to a spinning disk confocal... but who knows until we measure it.
It probably isnt as small/compact/tight as a properly adjusted point scanner with a closed pinhole.. but that doesn't detract from its utility.

Whether it makes sense to deconvolve such images remains to be verified theoretically and in practice.

Signal linearity... might well be an issue... as it is for apotome/optigrid - unless you really take care of and understand the details....
but the same is always true for any modality i guess.

just my 2 cents

Dan

Dr. Daniel James White BSc. (Hons.) PhD
Senior Microscopist / Image Visualisation, Processing and Analysis
Light Microscopy and Image Processing Facilities
Max Planck Institute of Molecular Cell Biology and Genetics
Pfotenhauerstrasse 108
01307 DRESDEN
Germany

+49 (0)15114966933 (German Mobile)
+49 (0)351 210 2627 (Work phone at MPI-CBG)
+49 (0)351 210 1078 (Fax MPI-CBG LMF)

http://www.bioimagexd.net BioImageXD
http://pacific.mpi-cbg.de Fiji - is just ImageJ (Batteries Included)
http://www.chalkie.org.uk Dan's Homepages
https://ifn.mpi-cbg.de Dresden Imaging Facility Network
dan (at) chalkie.org.uk
( white (at) mpi-cbg.de )

Guy Cox-2

Re: Vivatome experience

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

This is one of my pet hobby-horses, and many regular list members will know exactly what I'm going to say!

A confocal microscope only throws away out of focus light, which we don't want anyway, whatever form of microscopy we are doing. All the in focus light should go through the pinhole and end up in the final image. Why then do some people get brighter images with no obvious loss in resolution by opening the pinhole a bit? Because spherical aberration isn't properly corrected so they don't have an Airy disk at the pinhole, they have a circle of least confusion which is larger. Often this is their fault, but not always. It is virtually impossible to get an aberration-free image with a x40 NA0.75 lens however careful you are in preparation. If you don't believe me, try a comparison with the same manufacturer's x40 NA 0.95, which will have a correction collar - the difference is spectacular, and far greater than the difference in NA would suggest. (That assumes you know how to adjust the collar, of course). The same applies, to a lesser extent, with an NA 1.4 oil lens. You can fiddle with your Cargille oils, but a correction collar would make life much simpler.

Having said that, I don't deny that a confocal image has a worse S/N. But that's just statistics. In a 1 second image at 512x512, each point in the widefield image is recorded for 1s. In the equivalent confocal image it is recorded for ~4µs - there are a few orders of magnitude there! But it has nothing to do with throwing away light.

Guy

Sponsor my next half-marathon on May 15th

There's a special reason - find it out at

http://www.everydayhero.com.au/Guy_Cox_4846 <http://www.everydayhero.com.au/Guy_Cox_4846>

______________________________________________

Associate Professor Guy Cox, MA, DPhil(Oxon)

Australian Centre for Microscopy & Microanalysis,

Madsen Building F09, University of Sydney, NSW 2006

Phone +61 2 9351 3176 Fax +61 2 9351 7682

Mobile 0413 281 861

______________________________________________

http://www.guycox.net <http://www.guycox.net/>

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Daniel James White
Sent: Tuesday, 22 March 2011 7:21 PM
To: [hidden email]
Subject: Re: Vivatome experience

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Hi Jeremy,

On Mar 22, 2011, at 6:06 AM, CONFOCALMICROSCOPY automatic digest system wrote:

Keith Morris

Re: Cotton wool for lens cleaning

In reply to this post by Grant MacGregor

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

For a good discussion on microscope cleaning see:

http://micro.magnet.fsu.edu/primer/anatomy/cleaning.html

But then they pussyfoot around the issue as well - immersion oil damages
microscopes, solvents damage microscopes, cleaning damages microscopes, not
cleaning damages microscopes etc...all true I suppose. Molecular Expressions
seem to favour using bits of dead antelope [chamois leather] over 100%
cotton buds [just in case they have man-made fibres added], each to their
own.

Microscope engineers have been using ether for probably more than a hundred
years, and I've used it for 30 without any problems [although my 10ml will
evaporate from the bottle over the years before its used up, as its used so
sparingly]. My comments are really aimed at those of us that use oil
immersion objectives. Organic solvents shouldn't be used routinely on oil
immersion objectives either, perhaps a few times a year at best, and are
really a last resort when the objective is so dirty it's unusable. Removing
the immersion oil as part of a serious decontamination process should only
take a few seconds. I don't scrub, just touch the cotton bud on the lens
surface with a few gentle circular movements and then turn it over to the
other clean side and remove the solvent + oil.

That said soap and water removes major immersion oil spillages on our
removable glass slide holder stage-insert very effectively and whereas some
organic solvents would take the black paint off as well. One problem with
water based lens cleaners is if the stuff dries onto the glass, which can be
terminal for a lens [so I use ultra pure water, and OK a bit more ether to
ensure the waters removed fully, as alcohol is apparently less lens cement
friendly]. Unless the oil immersion objective has real contamination issues,
microscope tissues gently removing the excess of oil is all that is
required, day in day out. Our air objectives only require a air jet/blower
every now and then - no touching required [unless cross contaminated with
immersion oil]. If an objective seems to need a lot of cleaning, it's
probably damaged.

Most problems develop in multi-user facilities where oil and air objectives
are shared on an inverted microscope used for live cell imaging [for which
we tick all the boxes].

Regards

Keith

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford OX3 7BN,
United Kingdom.

Telephone: +44 (0)1865 287568
Email: [hidden email]
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Grant MacGregor
Sent: 21 March 2011 19:20
To: [hidden email]
Subject: Re: Cotton wool for lens cleaning

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I concur with Craig and Mark regarding use of ether and any other solvent of
lens cements. Many years ago I watched the front lens of a 1.4NA condenser
fall out following too much elbow grease with ether-soaked cotton buds /
Q-tips. Ether no more.

Several years ago someone dumped a plate full of tissue culture media down
the nosepiece of a shared facility's vert 200m I was in charge of
overseeing. Prior to fitting the aqua stop, I watched with great interest
to see what the extremely senior, German accented and fussy Zeiss field
service engineer would use to clean the optics. Would it be Opti L ? Would
it be pure ether ? Surprise surprise, he whipped out a large spray bottle
of Sparkle. This is a surfactant-laden ammonia free solution one can
purchase online or at an Ace Hardware (if in the US). The regular Sparkle
is purple. A while ago I suspect the makers of Sparkle found out that folks
were using it for high-end optics as they started packaging it in smaller
(c. 100ml) bottles and leaving out the purple dye. I don't know if they
charge more for this, but I would assume so. I discovered if you leave the
bottle of purple Sparkle in the window, the purple dye (presumably) oxidises
with a few weeks, becoming colorless.

For what it is worth, our general rule of thumb for cleaning optics is as
follows.

Try in order the following solvents - 1) distilled water; 2) Sparkle. If we
have to use Sparkle, we follow up with distilled water, which can be simply
applied by breathing on the optic.
Clean using only pure cotton or silk and not lens paper. I was taught that
lens paper should be used only for storage of optics. Back in the day,
microscope field engineers often wore silk ties that had a bunch of stains
on the back of them.
Clean in a circular manner in a single direction, taking care not to drag
any dirt back over the cleaned area.

Grant MacGregor
Developmental and Cell Biology,
UC Irvine

On Mar 17, 2011, at 12:04 PM, Craig Brideau wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I agree with Mark. Acetone will eat any rubbers/sealers/epoxies holding
> your lens together. I only use acetone on mirrors out on the optical
bench
> that are held in place with friction mounts (i.e. no epoxy). For anything
> glued or sealed I use high-purity methanol and lens tissues.
>
> Craig
>
>
> On Thu, Mar 17, 2011 at 7:09 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
>> *****
>>
>> From direct experience, I urge a note of caution with acetone, you may
>> degrade the lens mounting glue and risk spreading it over the lens.
>>
>> Regards Mark
>>
>>
>> On 17/03/2011, at 12:40 PM, Cammer, Michael wrote:
>>
>> *****
>>> To join, leave or search the confocal microscopy listserv, go to:
>>> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>> *****
>>>
>>> I don't think there really is one general solvent. When I began doing
>>> biological microscopy about 20 years ago the immersion oils all seemed

to be
>>> made of the same stuff and easily cleaned with just about any organic
>>> solvent stronger than ethanol. This is no longer the case. For
instance,
>>> the new Nikon oil for TIRF gets thick and is completely impervious to
any of
>>> the aqueous cleaners. It is resistant to what we practically considered
to
>>> be the universal solvent of organics, acetone, and also to ethanol. But
>>> dehydrated methanol works great. On the other hand, the Zeiss oils,
when
>>> fresh, clean up fine with their aqueous cleaning solutions and when old
and
>>> dripped all over the turret and such, with acetone. The old standby in
the
>>> lab, Cargill Labs type DF, cleans up with any inorganic solvent. Of
course,
>>> in one lab the gospel was xylene because, well, we scientists tend to be
>>> superstitious or traditional. As for ether, one benefit of using it, we
>>> were told years ago by someone at Zeiss, is that it evaporates so fast
that
>>> it reduces the chances of dissolving the glue holding in the front glass
of
>>> the objective. Is this really a problem? I've never had one of these
front
>>> lenses come loose. Now I tend to use 1:1 acetone/methanol and cotton
swabs
>>> and/or lens tissue following in the footsteps of Spectraphysics service
who
>>> uses this to clean their mirrors and gives us average power of a Watt
with
>>> 100 fs pulses at 910-920 nm, so I follow by example.
>>> -Michael Cammer
>>>
>>> ------------------------------------------------------------
>>> 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.
>>> =================================
>>>
>>
>

Keith Morris

Re: Cotton wool for lens cleaning

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Sorry I should have added the only reason I want the top lens of the
immersion oil objective cleaned free of immersion oil is so that I can look
though the objective [from the rear] via a magnifying glass and check that
there is nothing nasty deposited onto the top lens [or lenses elsewhere in
the barrel] affecting the objectives view. I'd would do this because I have
suspicions that the lens is contaminated [i.e. even my super nice test
slides look really bad down the microscope, and I've eliminated any other
likely reason for this].

Keith

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford OX3 7BN,
United Kingdom.

Telephone: +44 (0)1865 287568
Email: [hidden email]
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Keith Morris
Sent: 22 March 2011 12:28
To: [hidden email]
Subject: Re: Cotton wool for lens cleaning

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

For a good discussion on microscope cleaning see:

http://micro.magnet.fsu.edu/primer/anatomy/cleaning.html

But then they pussyfoot around the issue as well - immersion oil damages
microscopes, solvents damage microscopes, cleaning damages microscopes, not
cleaning damages microscopes etc...all true I suppose. Molecular Expressions
seem to favour using bits of dead antelope [chamois leather] over 100%
cotton buds [just in case they have man-made fibres added], each to their
own.

Microscope engineers have been using ether for probably more than a hundred
years, and I've used it for 30 without any problems [although my 10ml will
evaporate from the bottle over the years before its used up, as its used so
sparingly]. My comments are really aimed at those of us that use oil
immersion objectives. Organic solvents shouldn't be used routinely on oil
immersion objectives either, perhaps a few times a year at best, and are
really a last resort when the objective is so dirty it's unusable. Removing
the immersion oil as part of a serious decontamination process should only
take a few seconds. I don't scrub, just touch the cotton bud on the lens
surface with a few gentle circular movements and then turn it over to the
other clean side and remove the solvent + oil.

That said soap and water removes major immersion oil spillages on our
removable glass slide holder stage-insert very effectively and whereas some
organic solvents would take the black paint off as well. One problem with
water based lens cleaners is if the stuff dries onto the glass, which can be
terminal for a lens [so I use ultra pure water, and OK a bit more ether to
ensure the waters removed fully, as alcohol is apparently less lens cement
friendly]. Unless the oil immersion objective has real contamination issues,
microscope tissues gently removing the excess of oil is all that is
required, day in day out. Our air objectives only require a air jet/blower
every now and then - no touching required [unless cross contaminated with
immersion oil]. If an objective seems to need a lot of cleaning, it's
probably damaged.

Most problems develop in multi-user facilities where oil and air objectives
are shared on an inverted microscope used for live cell imaging [for which
we tick all the boxes].

Regards

Keith

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford OX3 7BN,
United Kingdom.

Telephone: +44 (0)1865 287568
Email: [hidden email]
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Grant MacGregor
Sent: 21 March 2011 19:20
To: [hidden email]
Subject: Re: Cotton wool for lens cleaning

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

I concur with Craig and Mark regarding use of ether and any other solvent of
lens cements. Many years ago I watched the front lens of a 1.4NA condenser
fall out following too much elbow grease with ether-soaked cotton buds /
Q-tips. Ether no more.

Several years ago someone dumped a plate full of tissue culture media down
the nosepiece of a shared facility's vert 200m I was in charge of
overseeing. Prior to fitting the aqua stop, I watched with great interest
to see what the extremely senior, German accented and fussy Zeiss field
service engineer would use to clean the optics. Would it be Opti L ? Would
it be pure ether ? Surprise surprise, he whipped out a large spray bottle
of Sparkle. This is a surfactant-laden ammonia free solution one can
purchase online or at an Ace Hardware (if in the US). The regular Sparkle
is purple. A while ago I suspect the makers of Sparkle found out that folks
were using it for high-end optics as they started packaging it in smaller
(c. 100ml) bottles and leaving out the purple dye. I don't know if they
charge more for this, but I would assume so. I discovered if you leave the
bottle of purple Sparkle in the window, the purple dye (presumably) oxidises
with a few weeks, becoming colorless.

For what it is worth, our general rule of thumb for cleaning optics is as
follows.

Try in order the following solvents - 1) distilled water; 2) Sparkle. If we
have to use Sparkle, we follow up with distilled water, which can be simply
applied by breathing on the optic.
Clean using only pure cotton or silk and not lens paper. I was taught that
lens paper should be used only for storage of optics. Back in the day,
microscope field engineers often wore silk ties that had a bunch of stains
on the back of them.
Clean in a circular manner in a single direction, taking care not to drag
any dirt back over the cleaned area.

Grant MacGregor
Developmental and Cell Biology,
UC Irvine

On Mar 17, 2011, at 12:04 PM, Craig Brideau wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I agree with Mark. Acetone will eat any rubbers/sealers/epoxies holding
> your lens together. I only use acetone on mirrors out on the optical
bench
> that are held in place with friction mounts (i.e. no epoxy). For anything
> glued or sealed I use high-purity methanol and lens tissues.
>
> Craig
>
>
> On Thu, Mar 17, 2011 at 7:09 AM, Mark Cannell
<[hidden email]>wrote:

Martin Wessendorf-2

Re: Cotton wool for lens cleaning

In reply to this post by JOEL B. SHEFFIELD

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

On 3/21/2011 6:29 PM, JOEL B. SHEFFIELD wrote:

> This has been a fascinating discussion. It occurs to me, though, that we
> may be trying to find the perfect cleaning method that will apply to many
> different kinds of contamination. I can think of several that seem to me to
> be mutually exclusive.
>
> 1. Water soluble --or once soluble. I am thinking of dried salts from
> medium, from bathing solutions, tears of operators, etc.
>
> 2. Oil, deliberate on oil immersion lenses, or accidental, from
> inadvertently dipping a dry lens where it shouldn't go.
>
> 3. Dried nail polish or other mounting media, inappropriately applied, or
> from slides viewed by impatient students (!!!).
>
> At any rate, each of these probably needs a different approach. Organic
> solvents won't do much for type 2, and "Sparkle" won't make type 3 sparkle.

Excellent points. I'd add one more consideration: when cleaning lenses,
"First do no harm".

--I vividly remember trying to clean a dark, tar-like substance from the
bottom of a flask in organic chemistry class. I scraped, washed with
acetone, scraped more, used more acetone, etc. etc. for probably 15
minutes trying to clean the flask. I got too vigorous with the
scraping, though, and broke the flask; the pieces fell into the sink. I
picked them up and saw that the water in the sink--plain WATER!--had
completely dissolved the stuff off the glass shards.

So perhaps the most conservative way to proceed would be:

A: Examine the lens to try to see what the problem is. The suggestion I
heard years ago, and that I've used since, is to pull the eyepiece off
the microscope and use that as a magnifier when examining a lens.

B: Having identified the problem, use an appropriate cleaning method.
--If it's mounting medium or nail varnish, let it dry--once dry,
removing the stuff requires no solvents. The trick that I got from our
Olympus dealer was to use a splinter of wood (e.g., from a the broken
end of an applicator stick or cotton swab) to chip it off. When I've
had to use this method, it's worked very well.
--If it's salts, glycerol or other water-soluble stuff, use distilled
water.
--If it's oil, try Sparkle or some Kodak lens cleaner.
--Very last resort: organic solvents.

C: Check the lens after each step to see if further cleaning is needed.

Martin
--
Martin Wessendorf, Ph.D. office: (612) 626-0145
Assoc Prof, Dept Neuroscience lab: (612) 624-2991
University of Minnesota Preferred FAX: (612) 624-8118
6-145 Jackson Hall, 321 Church St. SE Dept Fax: (612) 626-5009
Minneapolis, MN 55455 e-mail: [hidden email]

Keith Morris

Re: Cotton wool for lens cleaning

In reply to this post by JOEL B. SHEFFIELD

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Zeiss do have a very useful pdf of 'how to clean your microscope' called
'The clean microscope':

http://www.zeiss.de/C1256D18002CC306/0/1205E8CBD68054EAC125701500404705/$fil
e/50-1-0011_e.pdf

I've never seen 'Sparkle' before, as downtown Illinois isn't that convenient
for me ['Sparkle' used to be the name of a furniture wax polish over here],
but a quick search found the product:

http://www.bestcleaningproducts.com/shop/product.asp?prodID=3779

I am almost persuaded to give up my ether habit for it.

Regards

Keith

---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford OX3 7BN,
United Kingdom.

Telephone: +44 (0)1865 287568
Email: [hidden email]
Web-pages: http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of JOEL B. SHEFFIELD
Sent: 21 March 2011 23:30
To: [hidden email]
Subject: Re: Cotton wool for lens cleaning

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

This has been a fascinating discussion. It occurs to me, though, that we
may be trying to find the perfect cleaning method that will apply to many
different kinds of contamination. I can think of several that seem to me to
be mutually exclusive.

1. Water soluble --or once soluble. I am thinking of dried salts from
medium, from bathing solutions, tears of operators, etc.

2. Oil, deliberate on oil immersion lenses, or accidental, from
inadvertently dipping a dry lens where it shouldn't go.

3. Dried nail polish or other mounting media, inappropriately applied, or
from slides viewed by impatient students (!!!).

At any rate, each of these probably needs a different approach. Organic
solvents won't do much for type 2, and "Sparkle" won't make type 3 sparkle.

Joel

On Mon, Mar 21, 2011 at 6:19 PM, Kathryn Spencer
<[hidden email]>wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> I'll join in...
> Has anyone worked with methyl ethyl ketone? I use this only as a
> weapon of last resort, for really nasty, can't-otherwise-clean unknown
gunk
> on core scope objectives. I use VWR cotton swabs, with a minimal amount of
> MEK, staying away from any glue/plastic/non-glass surface. For routine
> cleaning (once a month), I use isopropanol on lens tissue. Is this bad?
> Kathy
> The Scripps Research Institute
>

--

Joel B. Sheffield, Ph.D
Department of Biology
Temple University
Philadelphia, PA 19122
Voice: 215 204 8839
e-mail: [hidden email]
URL: http://astro.temple.edu/~jbs

Gregg Sobocinski

Re: Cotton wool for lens cleaning -How about ethanol to remove wet oil?

In reply to this post by Martin Wessendorf-2

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

Anyone aware of any hazards associated with using ethanol to remove undried immersion oil from dry or glycerin lenses? Since Sparkle is an aqueous solution, I found I needed 2-3 passes to remove oil from the lenses, especially with hard lens paper. My past experience is that a single pass with even 50% ethanol takes the oil off, then I follow up with a swipe of Sparkle to remove the spots left by the ethanol.

This is indeed a good topic! I look forward to your replies.
~Gregg

Gregg Sobocinski
Microscope Imaging Specialist
University of Michigan, MCDB Dept.
Ann Arbor, Michigan
USA

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Martin Wessendorf
Sent: Tuesday, March 22, 2011 10:39 AM
To: [hidden email]
Subject: Re: Cotton wool for lens cleaning

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
*****

On 3/21/2011 6:29 PM, JOEL B. SHEFFIELD wrote: