Commercial solutions for calibration of FRAP units

[Anton Kamnev]

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Dear all,

In our LM core we face a problem of frequent calibration of our "FRAP"
devices. Although the calibration itself is quite fast, many users just
don't have good samples which could be used for this purpose.

Therefore, I was wondering if there are any commercial products for
calibration of bleaching devices? What I'm looking for is a slide with a
thin uniform layer of a dye which could be bleached. The dye should be
immobilized so a bleaching pattern would be observable for some period of
time after the bleaching.

I would really appreciate if you could share some experience about samples
you use for calibration of bleaching patterns.

Although I'm primarily interested in commercial solutions, I would very much
appreciate some protocols for home-made calibration slides as well.

Thanks in advance!
Anton.

---------------------------
Anton Kamnev, PhD
Imaging Manager
Mechanochemical Cell Biology Building
Division of Biomedical Cell Biology
Warwick Medical School
The University of Warwick
Coventry, CV4 7AL UK

tel: [phone]
cell: [phone]
email: [hidden email]

Divisional Secretary: Karl Waldron
Tel: [phone]
[hidden email]

[Jorge]

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We successfully used this product from Chroma (no commercial interest)

Autofluorescent Plastic Slides

(link https://www.chroma.com/products/filter-accessories/diagnostic-slides)

Hope that helps,

Jorge

2014-12-01 10:40 GMT-03:00 Anton Kamnev <[hidden email]>:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all,
>
> In our LM core we face a problem of frequent calibration of our "FRAP"
> devices. Although the calibration itself is quite fast, many users just
> don't have good samples which could be used for this purpose.
>
> Therefore, I was wondering if there are any commercial products for
> calibration of bleaching devices? What I'm looking for is a slide with a
> thin uniform layer of a dye which could be bleached. The dye should be
> immobilized so a bleaching pattern would be observable for some period of
> time after the bleaching.
>
> I would really appreciate if you could share some experience about samples
> you use for calibration of bleaching patterns.
>
> Although I'm primarily interested in commercial solutions, I would very
> much
> appreciate some protocols for home-made calibration slides as well.
>
> Thanks in advance!
> Anton.
>
> ---------------------------
> Anton Kamnev, PhD
> Imaging Manager
> Mechanochemical Cell Biology Building
> Division of Biomedical Cell Biology
> Warwick Medical School
> The University of Warwick
> Coventry, CV4 7AL UK
>
> tel: [phone]
> cell: [phone]
> email: [hidden email]
>
> Divisional Secretary: Karl Waldron
> Tel: [phone]
> [hidden email]
>

--
Jorge Toledo H
Ph.D
Biomedical Neuroscience Institute (BNI)
Laboratory of cellular and molecular neurobiology(Couve-Lab)
Laboratory of Scientific Image Analysis (SCIAN-Lab)
Faculty of Medicine
University of Chile
www.couvelab.org | www.scian.cl

[Csúcs Gábor-3]

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Dear Anton,

Just a quick idea: the plastic fluorescent slides distributed by Chroma
(but also others) can be also bleached (if you have a somewhat stronger
laser). Could this work for you?

Greetings    Gabor

On 12/1/14 2:40 PM, "Anton Kamnev" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>Post images on http://www.imgur.com and include the link in your posting.
>*****
>
>Dear all,
>
>In our LM core we face a problem of frequent calibration of our "FRAP"
>devices. Although the calibration itself is quite fast, many users just
>don't have good samples which could be used for this purpose.
>
>Therefore, I was wondering if there are any commercial products for
>calibration of bleaching devices? What I'm looking for is a slide with a
>thin uniform layer of a dye which could be bleached. The dye should be
>immobilized so a bleaching pattern would be observable for some period of
>time after the bleaching.
>
>I would really appreciate if you could share some experience about samples
>you use for calibration of bleaching patterns.
>
>Although I'm primarily interested in commercial solutions, I would very
>much
>appreciate some protocols for home-made calibration slides as well.
>
>Thanks in advance!
>Anton.
>
>---------------------------
>Anton Kamnev, PhD
>Imaging Manager
>Mechanochemical Cell Biology Building
>Division of Biomedical Cell Biology
>Warwick Medical School
>The University of Warwick
>Coventry, CV4 7AL UK
>
>tel: [phone]
>cell: [phone]
>email: [hidden email]
>
>Divisional Secretary: Karl Waldron
>Tel: [phone]
>[hidden email]

[Katherina Garcia Garcia]

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Hello,

We use blue ink (permanent marker) on a coverslip with a small drop of water on the slide. You can focus de ink and bleach it.

Regards,
Katherina



El 12/01/14, Anton Kamnev  <[hidden email]> escribió:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all,
>
> In our LM core we face a problem of frequent calibration of our "FRAP"
> devices. Although the calibration itself is quite fast, many users just
> don't have good samples which could be used for this purpose.
>
> Therefore, I was wondering if there are any commercial products for
> calibration of bleaching devices? What I'm looking for is a slide with a
> thin uniform layer of a dye which could be bleached. The dye should be
> immobilized so a bleaching pattern would be observable for some period of
> time after the bleaching.
>
> I would really appreciate if you could share some experience about samples
> you use for calibration of bleaching patterns.
>
> Although I'm primarily interested in commercial solutions, I would very much
> appreciate some protocols for home-made calibration slides as well.
>
> Thanks in advance!
> Anton.
>
> ---------------------------
> Anton Kamnev, PhD
> Imaging Manager
> Mechanochemical Cell Biology Building
> Division of Biomedical Cell Biology
> Warwick Medical School
> The University of Warwick
> Coventry, CV4 7AL UK
>
> tel: [phone]
> cell: [phone]
> email: [hidden email]
>
> Divisional Secretary: Karl Waldron
> Tel: [phone]
> [hidden email]

--
---------------------------------------------------------------
Katherina García García
Técnico Especialista en Microscopía y Citometría
Tlf. 954977432
[hidden email]
Centro Andaluz de Biología del Desarrollo
Consejo Superior de Investigaciones Científicas
Universidad Pablo Olavide
Ctra. Utrera km1
41013 Sevilla
España

[WAINWRIGHT James]

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*****

COMMERCIAL RESPONSE

Hi everyone,

Andor manufacture and supply "FRAPPA Calibration Slides" with our FRAPPA units.

They are thin gels of Ethidium Bromide. I'd imagine they're easy to make?

Our part code is XS-AND-001 and can be requested from your regional Microscopy Sales Engineers:

http://www.andor.com/map.aspx

I'm not sure of the cost for these, but I'd imagine it's pretty low. (I'm unable to provide costs because I am located in Technical Support and we have limited access to the price lists).

Best regards,

James

James Wainwright
Product Support Engineer (Microscopy Systems)
Europe, Middle East & Africa

Tel:   [phone]
Mob: [phone]
Fax: [phone]
Web:  andor.com/support



-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Anton Kamnev
Sent: 01 December 2014 13:41
To: [hidden email]
Subject: Commercial solutions for calibration of FRAP units

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear all,

In our LM core we face a problem of frequent calibration of our "FRAP"
devices. Although the calibration itself is quite fast, many users just don't have good samples which could be used for this purpose.

Therefore, I was wondering if there are any commercial products for calibration of bleaching devices? What I'm looking for is a slide with a thin uniform layer of a dye which could be bleached. The dye should be immobilized so a bleaching pattern would be observable for some period of time after the bleaching.

I would really appreciate if you could share some experience about samples you use for calibration of bleaching patterns.

Although I'm primarily interested in commercial solutions, I would very much appreciate some protocols for home-made calibration slides as well.

Thanks in advance!
Anton.

---------------------------
Anton Kamnev, PhD
Imaging Manager
Mechanochemical Cell Biology Building
Division of Biomedical Cell Biology
Warwick Medical School
The University of Warwick
Coventry, CV4 7AL UK

tel: [phone]
cell: [phone]
email: [hidden email]

Divisional Secretary: Karl Waldron
Tel: [phone]
[hidden email]

[Josef Gotzmann]

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Dear Anton,

we also don't use a commercial solution.
we just paint our coverslips with a highlighter pen (Stabilo Luminator XT) -
works perfect. Can also be used to test photoconversion - we have no details
on that, though the yellow highlighters converts into a orange/reddish
colour upon 405nm illumination.

best
Josef

On Mon, 1 Dec 2014 07:40:38 -0600, Anton Kamnev <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>Post images on http://www.imgur.com and include the link in your posting.
>*****
>
>Dear all,
>
>In our LM core we face a problem of frequent calibration of our "FRAP"
>devices. Although the calibration itself is quite fast, many users just
>don't have good samples which could be used for this purpose.
>
>Therefore, I was wondering if there are any commercial products for
>calibration of bleaching devices? What I'm looking for is a slide with a
>thin uniform layer of a dye which could be bleached. The dye should be
>immobilized so a bleaching pattern would be observable for some period of
>time after the bleaching.
>
>I would really appreciate if you could share some experience about samples
>you use for calibration of bleaching patterns.
>
>Although I'm primarily interested in commercial solutions, I would very much
>appreciate some protocols for home-made calibration slides as well.
>
>Thanks in advance!
>Anton.
>
>---------------------------
>Anton Kamnev, PhD
>Imaging Manager
>Mechanochemical Cell Biology Building
>Division of Biomedical Cell Biology
>Warwick Medical School
>The University of Warwick
>Coventry, CV4 7AL UK
>
>tel: [phone]
>cell: [phone]
>email: [hidden email]
>
>Divisional Secretary: Karl Waldron
>Tel: [phone]
>[hidden email]

[phil laissue-2]

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Donnert, Eggeling & Hell 2007 made a monolayer of Atto532 NHS ester on a
silanized coverslip, or alternately used purified GFP and Venus in a
~500-nm adsorbed layer (poly-L-lysine) and Mowiol-mounted.
http://www.ncbi.nlm.nih.gov/pubmed/17179937

_________________________________________
Philippe Laissue, PhD, Director of Bioimaging Unit
School of Biological Sciences, Room 4.17
University of Essex, Colchester CO4 3SQ, UK
(0044) 01206 872246 / (0044) 07842 676 456
[hidden email]
privatewww.essex.ac.uk/~plaissue

On 1 December 2014 at 13:40, Anton Kamnev <[hidden email]> wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> Post images on http://www.imgur.com and include the link in your posting.
> *****
>
> Dear all,
>
> In our LM core we face a problem of frequent calibration of our "FRAP"
> devices. Although the calibration itself is quite fast, many users just
> don't have good samples which could be used for this purpose.
>
> Therefore, I was wondering if there are any commercial products for
> calibration of bleaching devices? What I'm looking for is a slide with a
> thin uniform layer of a dye which could be bleached. The dye should be
> immobilized so a bleaching pattern would be observable for some period of
> time after the bleaching.
>
> I would really appreciate if you could share some experience about samples
> you use for calibration of bleaching patterns.
>
> Although I'm primarily interested in commercial solutions, I would very
> much
> appreciate some protocols for home-made calibration slides as well.
>
> Thanks in advance!
> Anton.
>
> ---------------------------
> Anton Kamnev, PhD
> Imaging Manager
> Mechanochemical Cell Biology Building
> Division of Biomedical Cell Biology
> Warwick Medical School
> The University of Warwick
> Coventry, CV4 7AL UK
>
> tel: [phone]
> cell: [phone]
> email: [hidden email]
>
> Divisional Secretary: Karl Waldron
> Tel: [phone]
> [hidden email]
>

[mcammer]

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

We've used these slides for this.
e.g. http://cammer.net/historical/aif/instructions/zeiss/liveduo/Zaxis_bleachingtest/index.htm
http://cammer.net/historical/aif/instructions/fluor-ref-slides/01.htm


=========================================================================
 Michael Cammer, Microscopy Core & Skirball Institute, NYU Langone Medical Center
                          Cell:  914-309-3270     Temporary location:  SK2-7
          http://ocs.med.nyu.edu/microscopy & http://microscopynotes.com/

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Csúcs Gábor
Sent: Monday, December 01, 2014 9:07 AM
To: [hidden email]
Subject: Re: Commercial solutions for calibration of FRAP units

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Anton,

Just a quick idea: the plastic fluorescent slides distributed by Chroma (but also others) can be also bleached (if you have a somewhat stronger laser). Could this work for you?

Greetings    Gabor

On 12/1/14 2:40 PM, "Anton Kamnev" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>Post images on http://www.imgur.com and include the link in your posting.
>*****
>
>Dear all,
>
>In our LM core we face a problem of frequent calibration of our "FRAP"
>devices. Although the calibration itself is quite fast, many users just
>don't have good samples which could be used for this purpose.
>
>Therefore, I was wondering if there are any commercial products for
>calibration of bleaching devices? What I'm looking for is a slide with
>a thin uniform layer of a dye which could be bleached. The dye should
>be immobilized so a bleaching pattern would be observable for some
>period of time after the bleaching.
>
>I would really appreciate if you could share some experience about
>samples you use for calibration of bleaching patterns.
>
>Although I'm primarily interested in commercial solutions, I would very
>much appreciate some protocols for home-made calibration slides as
>well.
>
>Thanks in advance!
>Anton.
>
>---------------------------
>Anton Kamnev, PhD
>Imaging Manager
>Mechanochemical Cell Biology Building
>Division of Biomedical Cell Biology
>Warwick Medical School
>The University of Warwick
>Coventry, CV4 7AL UK
>
>tel: [phone]
>cell: [phone]
>email: [hidden email]
>
>Divisional Secretary: Karl Waldron
>Tel: [phone]
>[hidden email]

[mcammer]

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

p.s.  Since these web pages over ten years ago, we added measuring the intensity at the sample with a power meter before doing anything like this.

=========================================================================
 Michael Cammer, Microscopy Core & Skirball Institute, NYU Langone Medical Center
                          Cell:  914-309-3270     Temporary location:  SK2-7
          http://ocs.med.nyu.edu/microscopy & http://microscopynotes.com/


-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Cammer, Michael
Sent: Monday, December 01, 2014 10:33 AM
To: [hidden email]
Subject: Re: Commercial solutions for calibration of FRAP units

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

We've used these slides for this.
e.g. http://cammer.net/historical/aif/instructions/zeiss/liveduo/Zaxis_bleachingtest/index.htm
http://cammer.net/historical/aif/instructions/fluor-ref-slides/01.htm


=========================================================================
 Michael Cammer, Microscopy Core & Skirball Institute, NYU Langone Medical Center
                          Cell:  914-309-3270     Temporary location:  SK2-7
          http://ocs.med.nyu.edu/microscopy & http://microscopynotes.com/

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Csúcs Gábor
Sent: Monday, December 01, 2014 9:07 AM
To: [hidden email]
Subject: Re: Commercial solutions for calibration of FRAP units

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear Anton,

Just a quick idea: the plastic fluorescent slides distributed by Chroma (but also others) can be also bleached (if you have a somewhat stronger laser). Could this work for you?

Greetings    Gabor

On 12/1/14 2:40 PM, "Anton Kamnev" <[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>Post images on http://www.imgur.com and include the link in your posting.
>*****
>
>Dear all,
>
>In our LM core we face a problem of frequent calibration of our "FRAP"
>devices. Although the calibration itself is quite fast, many users just
>don't have good samples which could be used for this purpose.
>
>Therefore, I was wondering if there are any commercial products for
>calibration of bleaching devices? What I'm looking for is a slide with
>a thin uniform layer of a dye which could be bleached. The dye should
>be immobilized so a bleaching pattern would be observable for some
>period of time after the bleaching.
>
>I would really appreciate if you could share some experience about
>samples you use for calibration of bleaching patterns.
>
>Although I'm primarily interested in commercial solutions, I would very
>much appreciate some protocols for home-made calibration slides as
>well.
>
>Thanks in advance!
>Anton.
>
>---------------------------
>Anton Kamnev, PhD
>Imaging Manager
>Mechanochemical Cell Biology Building
>Division of Biomedical Cell Biology
>Warwick Medical School
>The University of Warwick
>Coventry, CV4 7AL UK
>
>tel: [phone]
>cell: [phone]
>email: [hidden email]
>
>Divisional Secretary: Karl Waldron
>Tel: [phone]
>[hidden email]

[Lange, Jeffrey]

*****
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*****

Hello all-

I commonly mark the inside of a Mat Tek dish with a red sharpie marker. This gives a very flat uniform field that is easily imaged and bleached with a 561nm laser. There is no recovery of the marker over the course of months.

Jeff

-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Anton Kamnev
Sent: Monday, December 01, 2014 7:41 AM
To: [hidden email]
Subject: Commercial solutions for calibration of FRAP units

*****
To join, leave or search the confocal microscopy listserv, go to:
http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
Post images on http://www.imgur.com and include the link in your posting.
*****

Dear all,

In our LM core we face a problem of frequent calibration of our "FRAP"
devices. Although the calibration itself is quite fast, many users just don't have good samples which could be used for this purpose.

Therefore, I was wondering if there are any commercial products for calibration of bleaching devices? What I'm looking for is a slide with a thin uniform layer of a dye which could be bleached. The dye should be immobilized so a bleaching pattern would be observable for some period of time after the bleaching.

I would really appreciate if you could share some experience about samples you use for calibration of bleaching patterns.

Although I'm primarily interested in commercial solutions, I would very much appreciate some protocols for home-made calibration slides as well.

Thanks in advance!
Anton.

---------------------------
Anton Kamnev, PhD
Imaging Manager
Mechanochemical Cell Biology Building
Division of Biomedical Cell Biology
Warwick Medical School
The University of Warwick
Coventry, CV4 7AL UK

tel: [phone]
cell: [phone]
email: [hidden email]

Divisional Secretary: Karl Waldron
Tel: [phone]
[hidden email]

[Barbara Foster]

*****
To join, leave or search the confocal microscopy listserv, go to:
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Post images on http://www.imgur.com and include the link in your posting.
*****

There are also Fluor-Ref slides from us, at www.MicroscopyEducation.com

These are the slides originally invented by P C Cheng.

CAVEAT:  Commercial Interest
Barbara Foster, President & Chief Consultant
Microscopy/Microscopy Education*
www.MicroscopyEducation.com

*A subsidiary of The Microscopy & Imaging Place, Inc.
7101 Royal Glen Trail, Suite A
McKinney, TX 75070
P: 972-924-5310
F: 214-592-0277

MME is currently scheduling courses for now and through Jan 2015.
Call us today for a free training evaluation.


At 07:48 AM 12/1/2014, Jorge wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>Post images on http://www.imgur.com and include the link in your posting.
>*****
>
>We successfully used this product from Chroma (no commercial interest)
>
>Autofluorescent Plastic Slides
>
>(link https://www.chroma.com/products/filter-accessories/diagnostic-slides)
>
>Hope that helps,
>
>Jorge
>
>2014-12-01 10:40 GMT-03:00 Anton Kamnev <[hidden email]>:
>
> > *****
> > To join, leave or search the confocal microscopy listserv, go to:
> > http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> > Post images on http://www.imgur.com and include the link in your posting.
> > *****
> >
> > Dear all,
> >
> > In our LM core we face a problem of frequent calibration of our "FRAP"
> > devices. Although the calibration itself is quite fast, many users just
> > don't have good samples which could be used for this purpose.
> >
> > Therefore, I was wondering if there are any commercial products for
> > calibration of bleaching devices? What I'm looking for is a slide with a
> > thin uniform layer of a dye which could be bleached. The dye should be
> > immobilized so a bleaching pattern would be observable for some period of
> > time after the bleaching.
> >
> > I would really appreciate if you could share some experience about samples
> > you use for calibration of bleaching patterns.
> >
> > Although I'm primarily interested in commercial solutions, I would very
> > much
> > appreciate some protocols for home-made calibration slides as well.
> >
> > Thanks in advance!
> > Anton.
> >
> > ---------------------------
> > Anton Kamnev, PhD
> > Imaging Manager
> > Mechanochemical Cell Biology Building
> > Division of Biomedical Cell Biology
> > Warwick Medical School
> > The University of Warwick
> > Coventry, CV4 7AL UK
> >
> > tel: [phone]
> > cell: [phone]
> > email: [hidden email]
> >
> > Divisional Secretary: Karl Waldron
> > Tel: [phone]
> > [hidden email]
> >
>
>
>
>--
>Jorge Toledo H
>Ph.D
>Biomedical Neuroscience Institute (BNI)
>Laboratory of cellular and molecular neurobiology(Couve-Lab)
>Laboratory of Scientific Image Analysis (SCIAN-Lab)
>Faculty of Medicine
>University of Chile
>www.couvelab.org | www.scian.cl