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Cleaning lens.

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Ian Montgomery

Cleaning lens.

Cleaning objectives, I use the fluid recommended by the local Zeiss engineer, 90% methylated spirit and 10% isopropanol.

Ian.

Dr. Ian Montgomery,

Histotechnology,

I.B.L.S. Support Unit,

Thomson Building,

University of Glasgow,

Glasgow,

G12 8QQ.

Kim Anker Kristiansen

Membrane specific dye to be used in IHC

Dear Listers,

I am looking for a membrane specific dye to use together with antibodies in IHC staining. We would like to set up a system where we look at receptors associated with the cell membrane, hence the membrane dye.

We are doing cryo-sectioning, acetone fixation and TBS buffered immunostaining.

So my question is: What would be a good choice of dye to apply to the already immunostained tissue?

Cheers Kim

Kim A. Kristiansen
Post Doc.
Glostrup Research Center
Department of Clinical Research
Glostrup Hospital
Ndr Ringvej 69
DK-2600 Glostrup
Denmark
Tel: +45 25219914
E-mail: [hidden email]

Keith Morris

Re: Cleaning lens.

In reply to this post by Ian Montgomery

‘Zeiss cleaning mixture L’, which the engineer’s now use since diethyl ether has been withdrawn from their kit, is 90% by volume ‘benzoline’ [petroleum ether sometimes called medical alcohol] and 10% ‘isopropanol’ [2-proponal, dimethyl carbinol, 2-hydroxyproparne]. The bottle says ‘Clean the optics by moving in circles, slight pressure should be exerted on optics during cleaning’. Petroleum ether or spirit isn’t the same as the diethyl ether solvent/anaesthetic often used to clean objectives, but apparently it does the job for Zeiss optics.

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/cytogenetics/

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ian Montgomery
Sent: 30 March 2009 12:28
To: [hidden email]
Subject: Cleaning lens.

Cleaning objectives, I use the fluid recommended by the local Zeiss engineer, 90% methylated spirit and 10% isopropanol.

Ian.

Dr. Ian Montgomery,

Histotechnology,

I.B.L.S. Support Unit,

Thomson Building,

University of Glasgow,

Glasgow,

G12 8QQ.

Ian Montgomery

Re: Cleaning lens.

Keith,

Methylated spirit that’s what he said, although I still prefer and use ether when necessary.

Ian.

Dr. Ian Montgomery,

Histotechnology,

I.B.L.S. Support Unit,

Thomson Building,

University of Glasgow,

Glasgow,

G12 8QQ.

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Keith Morris
Sent: 30 March 2009 14:12
To: [hidden email]
Subject: Re: Cleaning lens.

Keith

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ian Montgomery
Sent: 30 March 2009 12:28
To: [hidden email]
Subject: Cleaning lens.

Cleaning objectives, I use the fluid recommended by the local Zeiss engineer, 90% methylated spirit and 10% isopropanol.

Ian.

Dr. Ian Montgomery,

Histotechnology,

I.B.L.S. Support Unit,

Thomson Building,

University of Glasgow,

Glasgow,

G12 8QQ.

Fredrik Wermeling

Re: Membrane specific dye to be used in IHC

In reply to this post by Kim Anker Kristiansen

>
>
>
> We are doing cryo-sectioning, acetone fixation and TBS buffered
> immunostaining.
>
>
>
> So my question is: What would be a good choice of dye to apply to the
> already immunostained tissue?
>
>
>
> Cheers Kim
>
>
>
> Kim A. Kristiansen
> Post Doc.
> Glostrup Research Center
> Department of Clinical Research
> Glostrup Hospital
> Ndr Ringvej 69
> DK-2600 Glostrup
> Denmark
> Tel: +45 25219914
> E-mail: [hidden email]
>
>
>
>

Ward, Christopher W. (SON)

Re: Membrane specific dye to be used in IHC

We use a WGA(wheat germ agglutinin) - ALEXA conjugate (Invitrogen) in muscle cross-sections.

We apply this after our primary and secondary and get beautiful results.

___________________________________________

Christopher W. Ward Ph.D.

Associate Professor

University of Maryland Baltimore: School of Nursing

655 W. Lombard St.

Rm 752

Baltimore MD 21201

Ph: 410.706.3618

Fax: 410.706.0399

Skype: chrisward_umbaltimore

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Fredrik Wermeling
Sent: Monday, March 30, 2009 10:44 AM
To: [hidden email]
Subject: Re: Membrane specific dye to be used in IHC

I know that people use cholera-toxin B-subunit conjugates to label lipid rafts. I think you can find it conjugated to several alexa dyes. /fredrik

--------------------------------------
Fredrik Wermeling, PhD-student
Karolinska Institutet
Department of Medicine
Unit of Clinical Allergy Research
L2:04 Karolinska Hospital
SE-171 76 Stockholm
Sweden

phone: +46-8-51776696
mail: [hidden email]
fax: +46-8-335724
--------------------------------------

----- Original Message -----
From: Kim Anker Kristiansen <[hidden email]>
Date: Monday, March 30, 2009 3:07 pm
Subject: Membrane specific dye to be used in IHC
To: [hidden email]

> Dear Listers,
>
>
>
> I am looking for a membrane specific dye to use together with
> antibodiesin IHC staining. We would like to set up a system where
> we look at
> receptors associated with the cell membrane, hence the membrane dye.
>
>
>
> We are doing cryo-sectioning, acetone fixation and TBS buffered
> immunostaining.
>
>
>
> So my question is: What would be a good choice of dye to apply to the
> already immunostained tissue?
>
>
>
> Cheers Kim
>
>
>
> Kim A. Kristiansen
> Post Doc.
> Glostrup Research Center
> Department of Clinical Research
> Glostrup Hospital
> Ndr Ringvej 69
> DK-2600 Glostrup
> Denmark
> Tel: +45 25219914
> E-mail: [hidden email]
>
>
>
>

Chris Tully

Re: Cleaning lens.

In reply to this post by Ian Montgomery

Dear all,

While working for a Leica Microsystems dealer the local field service engineer (factor trained) used a sequence of ethanol and heptane to clean truly dirty lenses. For standard cleaning a lens wipe and Sparkle was his recommendation. But for dried oil or the like he would graduate to cotton swabs and either ethanol then heptane or a 50:50 mix of the two.

Chris

Chris Tully
Microscopy and Image Analysis Expert
[hidden email]
240-888-1021
http://www.linkedin.com/in/christully

On Mon, Mar 30, 2009 at 10:12 AM, Ian Montgomery <[hidden email]> wrote:

Keith,

            Methylated spirit that’s what he said, although I still prefer and use ether when necessary.

Ian.

Dr. Ian Montgomery,

Histotechnology,

I.B.L.S. Support Unit,

Thomson Building,

University of Glasgow,

Glasgow,

G12 8QQ.

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Keith Morris
Sent: 30 March 2009 14:12

To: [hidden email]

Subject: Re: Cleaning lens.

Are you sure the Zeiss Engineer didn’t say ‘petroleum spirit’? Methylated spirit is mainly ethanol, and so best avoided - the Axiovert 100 manual says repeated use of 70% ethanol will damage the objectives [but you can use it if you want]. Generally the faster the solvent evaporation from the lens/cement area the better, hence the suggestion of the solvent [pure] diethyl ether by many [and that’s what I use].

‘Zeiss cleaning mixture L’, which the engineer’s now use since diethyl ether has been withdrawn from their kit, is 90% by volume ‘benzoline’ [petroleum ether sometimes called medical alcohol] and 10% ‘isopropanol’ [2-proponal, dimethyl carbinol, 2-hydroxyproparne]. The bottle says ‘Clean the optics by moving in circles, slight pressure should be exerted on optics during cleaning’. Petroleum ether or spirit isn’t the same as the diethyl ether solvent/anaesthetic often used to clean objectives, but apparently it does the job for Zeiss optics.

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/cytogenetics/

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ian Montgomery
Sent: 30 March 2009 12:28
To: [hidden email]
Subject: Cleaning lens.

            In one of our teaching labs many years ago a student complained they were having a problem with the x100 OI objective and sure enough the image was lousy. I cleaned the objective and slide then re-applied a spot of oil and still the image was lousy. I then asked the student how exactly they had set up the microscope. Shock horror, my world collapsed. They had unscrewed the objective, filled it with oil, screwed it back on then put a drop on the slide. After weeks of trying to clean the objective it went into the trash as beyond economic repair.

            Cleaning objectives, I use the fluid recommended by the local Zeiss engineer, 90% methylated spirit and 10% isopropanol.

Ian.

Dr. Ian Montgomery,

Histotechnology,

I.B.L.S. Support Unit,

Thomson Building,

University of Glasgow,

Glasgow,

G12 8QQ.

Jean-Pierre CLAMME-2

Re: Membrane specific dye to be used in IHC

In reply to this post by Ward, Christopher W. (SON)

Is WGA specific to plasma membrane ?

Confocal Microscopy List <[hidden email]> wrote on 03/30/2009 07:47:09 AM: > We use a WGA(wheat germ agglutinin) - ALEXA conjugate (Invitrogen) > in muscle cross-sections.
> > We apply this after our primary and secondary and get beautiful results.
> >
> >
> >
> >
> >
> >
> >
> >
> > ___________________________________________
> > Christopher W. Ward Ph.D.
> > Associate Professor
> > University of Maryland Baltimore: School of Nursing
> > 655 W. Lombard St.
> > Rm 752
> > Baltimore MD 21201
> >
> > Ph: 410.706.3618
> > Fax: 410.706.0399
> > Skype: chrisward_umbaltimore
> >
> >
> >
> > From:Confocal Microscopy List [mailto:[hidden email]] > On Behalf Of Fredrik Wermeling
> Sent: Monday, March 30, 2009 10:44 AM
> To: [hidden email]
> Subject: Re: Membrane specific dye to be used in IHC
> >
> > I know that people use cholera-toxin B-subunit conjugates to label > lipid rafts. I think you can find it conjugated to several alexa > dyes. /fredrik
> > --------------------------------------
> Fredrik Wermeling, PhD-student
> Karolinska Institutet
> Department of Medicine
> Unit of Clinical Allergy Research
> L2:04 Karolinska Hospital
> SE-171 76 Stockholm
> Sweden
> > phone: +46-8-51776696
> mail: [hidden email]
> fax: +46-8-335724
> --------------------------------------
> > ----- Original Message -----
> From: Kim Anker Kristiansen <[hidden email]>
> Date: Monday, March 30, 2009 3:07 pm
> Subject: Membrane specific dye to be used in IHC
> To: [hidden email]
> > > Dear Listers,
> >
> >
> >
> > I am looking for a membrane specific dye to use together with
> > antibodiesin IHC staining. We would like to set up a system where
> > we look at
> > receptors associated with the cell membrane, hence the membrane dye.
> >
> >
> >
> > We are doing cryo-sectioning, acetone fixation and TBS buffered
> > immunostaining.
> >
> >
> >
> > So my question is: What would be a good choice of dye to apply to the
> > already immunostained tissue?
> >
> >
> >
> > Cheers Kim
> >
> >
> >
> > Kim A. Kristiansen
> > Post Doc.
> > Glostrup Research Center
> > Department of Clinical Research
> > Glostrup Hospital
> > Ndr Ringvej 69
> > DK-2600 Glostrup
> > Denmark
> > Tel: +45 25219914
> > E-mail: [hidden email]
> >
> >
> >
> >

JOEL B. SHEFFIELD

Re: Membrane specific dye to be used in IHC

Actually, WGA binds to specific glycoproteins, and so it will
frequently bind to the glycoproteins in membranes. However, it will
also bind to Golgi membranes, but in some cases, where the
glycoprotein array is of different composition, it won't bind at all.
In those cases you might find that another lectin, such as ConA,
will be better. In all of these situations, though, the lectins will
also bind to other glycosylated components of the extracellular
matrix, so some caution is indicated.

Ideally, you might be interested in a lipid-binding compound, such as
FM 4-64. However, since your procedure extracts most lipids, this
may not be a useful approach.

Joel

-------------- Original message ---------------

Is WGA specific to plasma membrane ?

Confocal Microscopy List <[hidden email]> wrote on 03/30/2009 07:47:09 AM:

> We use a WGA(wheat germ agglutinin) - ALEXA conjugate (Invitrogen)
> in muscle cross-sections.
>
> We apply this after our primary and secondary and get beautiful results.
>
>
>
>
>
>
>
>
>
>
>
>
>
>
>
>
>
> ___________________________________________
>
> Christopher W. Ward Ph.D.
>
> Associate Professor
>
> University of Maryland Baltimore: School of Nursing
>
> 655 W. Lombard St.
>
> Rm 752
>
> Baltimore MD 21201
>
>
>
> Ph: 410.706.3618
>
> Fax: 410.706.0399
>
> Skype: chrisward_umbaltimore
>
>
>
>
>
>
>
> From:Confocal Microscopy List [mailto:[hidden email]]
> On Behalf Of Fredrik Wermeling
> Sent: Monday, March 30, 2009 10:44 AM
> To: [hidden email]
> Subject: Re: Membrane specific dye to be used in IHC
>
>
>
> I know that people use cholera-toxin B-subunit conjugates to label
> lipid rafts. I think you can findit conjugated to severalalexa
> dyes. /fredrik
>
> --------------------------------------
> Fredrik Wermeling, PhD-student
> Karolinska Institutet
> Department of Medicine
> Unit of Clinical Allergy Research
> L2:04 Karolinska Hospital
> SE-171 76 Stockholm
> Sweden
>
> phone: +46-8-51776696
> mail: [hidden email]
> fax: +46-8-335724
>--------------------------------------
>
> ----- Original Message -----
> From: Kim Anker Kristiansen <[hidden email]>
> Date: Monday, March 30, 2009 3:07 pm
> Subject: Membrane specific dye to be used in IHC
> To: [hidden email]
>
> > Dear Listers,
> >
> >
> >
> > I am looking for a membrane specific dye to use together with
> > antibodiesin IHC staining. We would like to set up a system where
> > we look at
> > receptors associated with the cell membrane, hence the membrane dye.
> >
> >
> >
> > We are doing cryo-sectioning, acetone fixation and TBS buffered
> > immunostaining.
> >
> >
> >
> > So my question is: What would be a good choice of dye to apply to the
> > already immunostained tissue?
> >
> >
> >
> > Cheers Kim
> >
> >
> >
> > Kim A. Kristiansen
> > Post Doc.
> > Glostrup Research Center
> > Department of Clinical Research
> > Glostrup Hospital
> > Ndr Ringvej 69
> > DK-2600 Glostrup
> > Denmark
> > Tel: +45 25219914
> > E-mail: [hidden email]
> >
> >
> >
> >

--
Joel B. Sheffield, Ph.D.
Biology Department, Temple University
1900 North 12th Street
Philadelphia, PA 19122
[hidden email]
(215) 204 8839, fax (215) 204 0486
http://astro.temple.edu/~jbs

Neil Kad

Re: Cleaning lens.

In reply to this post by Chris Tully

Dear All,

I found this article on the Olympus website (no commercial interest):

www.olympus.co.uk/microscopy/images/illum_cleaning.pdf

I've used a cotton swab with methanol without any problems, yet.... Based on this discussion I might move to an ether/MeOH mixture!

Cheers

Neil

Date: Mon, 30 Mar 2009 11:39:36 -0400
From: [hidden email]
Subject: Re: Cleaning lens.
To: [hidden email]

Dear all,

While working for a Leica Microsystems dealer the local field service engineer (factor trained) used a sequence of ethanol and heptane to clean truly dirty lenses. For standard cleaning a lens wipe and Sparkle was his recommendation. But for dried oil or the like he would graduate to cotton swabs and either ethanol then heptane or a 50:50 mix of the two.

Chris

Chris Tully
Microscopy and Image Analysis Expert
[hidden email]
240-888-1021
http://www.linkedin.com/in/christully

On Mon, Mar 30, 2009 at 10:12 AM, Ian Montgomery <[hidden email]> wrote:

Keith,
            Methylated spirit that’s what he said, although I still prefer and use ether when necessary.
Ian.

Dr. Ian Montgomery,
Histotechnology,
I.B.L.S. Support Unit,
Thomson Building,
University of Glasgow,
Glasgow,
G12 8QQ.

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Keith Morris
Sent: 30 March 2009 14:12

To: [hidden email]

Subject: Re: Cleaning lens.

Are you sure the Zeiss Engineer didn’t say ‘petroleum spirit’? Methylated spirit is mainly ethanol, and so best avoided - the Axiovert 100 manual says repeated use of 70% ethanol will damage the objectives [but you can use it if you want]. Generally the faster the solvent evaporation from the lens/cement area the better, hence the suggestion of the solvent [pure] diethyl ether by many [and that’s what I use].

‘Zeiss cleaning mixture L’, which the engineer’s now use since diethyl ether has been withdrawn from their kit, is 90% by volume ‘benzoline’ [petroleum ether sometimes called medical alcohol] and 10% ‘isopropanol’ [2-proponal, dimethyl carbinol, 2-hydroxyproparne]. The bottle says ‘Clean the optics by moving in circles, slight pressure should be exerted on optics during cleaning’. Petroleum ether or spirit isn’t the same as the diethyl ether solvent/anaesthetic often used to clean objectives, but apparently it does the job for Zeiss optics.

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/cytogenetics/

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ian Montgomery
Sent: 30 March 2009 12:28
To: [hidden email]
Subject: Cleaning lens.

            In one of our teaching labs many years ago a student complained they were having a problem with the x100 OI objective and sure enough the image was lousy. I cleaned the objective and slide then re-applied a spot of oil and still the image was lousy. I then asked the student how exactly they had set up the microscope. Shock horror, my world collapsed. They had unscrewed the objective, filled it with oil, screwed it back on then put a drop on the slide. After weeks of trying to clean the objective it went into the trash as beyond economic repair.
            Cleaning objectives, I use the fluid recommended by the local Zeiss engineer, 90% methylated spirit and 10% isopropanol.
Ian.

Dr. Ian Montgomery,
Histotechnology,
I.B.L.S. Support Unit,
Thomson Building,
University of Glasgow,
Glasgow,
G12 8QQ.

Share your photos with Windows Live Photos – Free. Try it Now!

Wiegraebe, Winfried-2

Re: Cleaning lens.

We recommend to users of our facility:

- start exhaling at your lens and use polyester swaps.

- if this does not work, use Zeiss lens cleaner (you can get it free from your rep)

- if this does not work use

85% petroleum ether + 15% isopropanol.

This seams to work very well.

We have compiled some additional information about cleaning a microscope at

http://research.stowers-institute.org/wiw/external/Maintenance/cleaning.htm

Winfried

=======================================
Winfried Wiegraebe
Director, Stowers Microscopy Center
Stowers Institute for Medical Research
1000 East 50th Street, Kansas City, MO 64110

http://research.stowers-institute.org/wiw/index.htm
[hidden email]
816-926-4415

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Neil Kad
Sent: Tuesday, March 31, 2009 4:04 AM
To: [hidden email]
Subject: Re: Cleaning lens.

On Mon, Mar 30, 2009 at 10:12 AM, Ian Montgomery <[hidden email]> wrote:

Keith,
            Methylated spirit that’s what he said, although I still prefer and use ether when necessary.

Ian.

Dr. Ian Montgomery,
Histotechnology,
I.B.L.S. Support Unit,
Thomson Building,
University of Glasgow,
Glasgow,
G12 8QQ.

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Keith Morris
Sent: 30 March 2009 14:12

To: [hidden email]
Subject: Re: Cleaning lens.

Are you sure the Zeiss Engineer didn’t say ‘petroleum spirit’? Methylated spirit is mainly ethanol, and so best avoided - the Axiovert 100 manual says repeated use of 70% ethanol will damage the objectives [but you can use it if you want]. Generally the faster the solvent evaporation from the lens/cement area the better, hence the suggestion of the solvent [pure] diethyl ether by many [and that’s what I use].

‘Zeiss cleaning mixture L’, which the engineer’s now use since diethyl ether has been withdrawn from their kit, is 90% by volume ‘benzoline’ [petroleum ether sometimes called medical alcohol] and 10% ‘isopropanol’ [2-proponal, dimethyl carbinol, 2-hydroxyproparne]. The bottle says ‘Clean the optics by moving in circles, slight pressure should be exerted on optics during cleaning’. Petroleum ether or spirit isn’t the same as the diethyl ether solvent/anaesthetic often used to clean objectives, but apparently it does the job for Zeiss optics.

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/cytogenetics/

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ian Montgomery
Sent: 30 March 2009 12:28
To: [hidden email]
Subject: Cleaning lens.

            In one of our teaching labs many years ago a student complained they were having a problem with the x100 OI objective and sure enough the image was lousy. I cleaned the objective and slide then re-applied a spot of oil and still the image was lousy. I then asked the student how exactly they had set up the microscope. Shock horror, my world collapsed. They had unscrewed the objective, filled it with oil, screwed it back on then put a drop on the slide. After weeks of trying to clean the objective it went into the trash as beyond economic repair.
            Cleaning objectives, I use the fluid recommended by the local Zeiss engineer, 90% methylated spirit and 10% isopropanol.
Ian.

Dr. Ian Montgomery,
Histotechnology,
I.B.L.S. Support Unit,
Thomson Building,
University of Glasgow,
Glasgow,
G12 8QQ.

Share your photos with Windows Live Photos – Free. Try it Now!

Williams, Geoffrey

Re: Cleaning lens.

I haven’t read the method that we train our users to follow so I’ll toss it into the mix.

(this is generally for inverted microscopes – upright stands don’t have to worry about leaving oil on the objective as much because they will generally not collect as much dust and cannot run down into the objective itself or on the microscope – for an upright just wiping the excess oil off and cleaning occasionally is usually sufficient)

Three things are needed:

1) a box (not an envelop – a box) of lens tissue, Specifically the Ross 4 ¼” x 5”. There are 1000 sheets per box and with daily use by many users each box lasts a good long time. Reason for the box? If there is going to be debris from the air falling on it the underside will always be clean. Since you pull a fresh piece off each time providing only the underside touches the objective lens you greatly reduce the risk of scratching it. I abhor the booklets. People never pull sheets out in order, invariably someone touches both sides of the tissue to remove it, using them to clean objectives gives me chills.

2) Puritan Cotton Tipped applicators (single end). Yes the long wooden applicators. These come in nice paper packages, when opened at the wood end the cotton side stays hidden and protected. The longer reach (than say a cotton swab) makes it easier for users (and reduces the temptation to flip the cotton swab around and use the end you were just holding). We thought we would try the Fisher Sci version of these (they were half the cost) and unfortunately they have about ¼ the amount of cotton winding as the Puritan Branded ones. You don’t need the double end but the Fisher brand ones barely have anything on them. Rather useless.

3) Sparkle Glass cleaner. Ammonia free is important, basically any ammonia free cleaner (I would avoid the vinegar enhanced ones). Or at least the importance was impressed on me that it could potentially remove the coating on the objective. I’m less concerned about the cement holding the objective lens in than I am of removing any coating on the objective glass itself. You can order this stuff direct (http://www.glasscleaner.com/ ) they have a Microscope lens cleaner but it just seems to be an expensive dye free version of the regular glass cleaner, but there are choices. I personally like having the purple stuff because it is a visual cue that you are using something different and specific. Having a clear solution I would be more concerned with someone using it on the water immersion objective or using water to clean the oil (words on a label aren’t always read).

Procedure:

Take one sheet of lens tissue out. Hold it flat by a corner. Dip the Cotton applicator into the bottle of Sparkle. Then place (the clean side towards the objective) a section of the lens tissue close to your fingers holding the tissue on the objective. Then with the damp/wet (not dripping) applicator apply a slight amount of force and drag the clean side of the tissue across the objective. Repeat until there is no more oil on the objective. You can tell it is clean by turning the paper over and looking for the shiny tell tale streak of oil. When the last few damp passes dry the tissue shouldn’t be shiny. Occasionally it takes more than one pass to clean. Again, this is for inverted microscopes.

Dry Objectives that get oil on them? Sometimes it is easy. But then I just go right to the cotton applicators and Sparkle, very gently cleaning and always getting new applicator. Often I will resort to plain old Q-tip swabs for cleaning dry objectives. Holding them in the middle get one end wet, ever so lightly swirl the sparkle over it, flip it around and dry, and repeat until the surface is completely clean under magnified inspection. If something is very stubborn but moving around (hard stuff like mounting medium I have fortunately not encountered on objectives here) I will soak just the very tip of the objective in a bit of full strength sparkle for a period of time (10-15 minutes or more), then clean with the swabs, and inspect and repeat until it is nice and clean. So far that method has served me quite well in the ~14-15 years I’ve been cleaning oily dry objectives.

Geoff Williams

Leduc Bioimaging Facility Manager

Brown University

http://www.brown.edu/Facilities/Leduc_Bioimaging_Facility/

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Neil Kad
Sent: Tuesday, March 31, 2009 4:04 AM
To: [hidden email]
Subject: Re: Cleaning lens.

Dear All,

I found this article on the Olympus website (no commercial interest):

www.olympus.co.uk/microscopy/images/illum_cleaning.pdf

I've used a cotton swab with methanol without any problems, yet.... Based on this discussion I might move to an ether/MeOH mixture!

Cheers

Neil

On Mon, Mar 30, 2009 at 10:12 AM, Ian Montgomery <[hidden email]> wrote:

Keith,
Methylated spirit that’s what he said, although I still prefer and use ether when necessary.

Ian.

Dr. Ian Montgomery,
Histotechnology,
I.B.L.S. Support Unit,
Thomson Building,
University of Glasgow,
Glasgow,
G12 8QQ.

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Keith Morris
Sent: 30 March 2009 14:12

To: [hidden email]

Subject: Re: Cleaning lens.

Are you sure the Zeiss Engineer didn’t say ‘petroleum spirit’? Methylated spirit is mainly ethanol, and so best avoided - the Axiovert 100 manual says repeated use of 70% ethanol will damage the objectives [but you can use it if you want]. Generally the faster the solvent evaporation from the lens/cement area the better, hence the suggestion of the solvent [pure] diethyl ether by many [and that’s what I use].

‘Zeiss cleaning mixture L’, which the engineer’s now use since diethyl ether has been withdrawn from their kit, is 90% by volume ‘benzoline’ [petroleum ether sometimes called medical alcohol] and 10% ‘isopropanol’ [2-proponal, dimethyl carbinol, 2-hydroxyproparne]. The bottle says ‘Clean the optics by moving in circles, slight pressure should be exerted on optics during cleaning’. Petroleum ether or spirit isn’t the same as the diethyl ether solvent/anaesthetic often used to clean objectives, but apparently it does the job for Zeiss optics.

Keith

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ian Montgomery
Sent: 30 March 2009 12:28
To: [hidden email]
Subject: Cleaning lens.

            In one of our teaching labs many years ago a student complained they were having a problem with the x100 OI objective and sure enough the image was lousy. I cleaned the objective and slide then re-applied a spot of oil and still the image was lousy. I then asked the student how exactly they had set up the microscope. Shock horror, my world collapsed. They had unscrewed the objective, filled it with oil, screwed it back on then put a drop on the slide. After weeks of trying to clean the objective it went into the trash as beyond economic repair.
            Cleaning objectives, I use the fluid recommended by the local Zeiss engineer, 90% methylated spirit and 10% isopropanol.
Ian.

Dr. Ian Montgomery,
Histotechnology,
I.B.L.S. Support Unit,
Thomson Building,
University of Glasgow,
Glasgow,
G12 8QQ.

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Dan Focht

Re: Cleaning lens.

Confocs

While this thread is still active I'll take this opportunity to remind you that after you clean an optic with an oil solvent make sure that you also clean it with a water based cleaner then distilled water. Often times the petroleum solvent will not remove sugars and salts that may have been left behind from media or fingers. The oil covers it up and you don't notice it but the image suffers.

Also make sure that all oil is removed from an objective especially after it has sat around for a while before applying more oil. If a thin film of oil is left on the objective overnight or over a weekend the oxidation changes the RI and viscosity of the oil leaving an irregular RI surface. If new oil is then added to this irregular surface the image is diminished. In my former life, when I serviced scopes, I often had customers that religiously blotted their immersion objectives with lens paper at the end of the day leaving behind a thin oil smear layer. They then added more oil to view a slide the following day. After a proper cleaning of the lens with petroleum solvent then water based cleaner and distilled water they were amazed at the improvement.

I believe the distilled water is a necessity. Take whatever glass cleaner you choose and let it evaporate on a mirror surface. It will leave a residue. The distilled water is necessary to remove this residue and leave a pure glass to air or glass to oil interface.

Dan

From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Ian Montgomery
Sent: 30 March 2009 12:28
To: [hidden email]
Subject: Cleaning lens.

            In one of our teaching labs many years ago a student complained they were having a problem with the x100 OI objective and sure enough the image was lousy. I cleaned the objective and slide then re-applied a spot of oil and still the image was lousy. I then asked the student how exactly they had set up the microscope. Shock horror, my world collapsed. They had unscrewed the objective, filled it with oil, screwed it back on then put a drop on the slide. After weeks of trying to clean the objective it went into the trash as beyond economic repair.
            Cleaning objectives, I use the fluid recommended by the local Zeiss engineer, 90% methylated spirit and 10% isopropanol.
Ian.

Dr. Ian Montgomery,
Histotechnology,
I.B.L.S. Support Unit,
Thomson Building,
University of Glasgow,
Glasgow,
G12 8QQ.

Share your photos with Windows Live Photos – Free. Try it Now!

Dan Focht

Bioptechs

724-282-7145

www.bioptechs.com

Coleman, Lorraine - Technician

Surface of a cloth

Hi All,

A question in relation to setting up a cloth to image in both a fluorescence and confocal microscope. The surface of a cloth should have FITC labelled antibodies on it and I am just wondering the best way to prepare the sample for imaging purposes, attach to a glass slide and mount a cover slip on top?

Any protocols/advice would be greatly appreciated.

Thanks

Kindest regards,

Lorraine.