Looking for recipe to create an Andor FRAPPA calibration slide?

[Wendy Salmon]

*****
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.
*****

Hi all,
Before I shell out cash to buy one, has anyone reverse-engineered the Andor
FRAPPA calibration slide and would be willing to share?  It's the smear of
Ethidium Bromide doped agarose (not the front surface mirror for the
MicroPoint system).
Thank you!
Wishing you peace and good health,
Wendy

--------------------
Wendy C Salmon, M.A. (she/her/hers)
Manager, W.M. Keck Facility for Biological Imaging

Whitehead Institute for Biomedical Research
455 Main St., Rm 447, Cambridge, MA 02142 USA
e: [hidden email]  |  w: microscopy.wi.mit.edu

<http://microscopy.wi.mit.edu>

[Ann-W]

*****
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.
*****

Hi Wendy

Yes, Well sort of. I used fluorescently labelled fibronectin and it did a very similar job. Although it was ages ago, I have a mosaic on my Andor system now not a FRAPPA. Essentially you need a fluorescent thing you can make a hole in and then calibrate the xy galvos of the FRAPPA to the position of the hole if my memory serves me correctly. I'm sure flurosecently labelled agarose would do the same job. The Fluorescently labelled Fibronectin can be brought from Thermo for about £350.

Hope this helps

Ann

Advanced Imaging Resource, Institute of Genetics and Molecular Medicine, University of Edinburgh, Edinburgh EH4 2XU
Working Pattern. Mon 9am - 2pm, Tues - Thurs 9am - 5pm.
E: [hidden email]
T: 0131 651 8665
W: http://www.ed.ac.uk/igmm-imaging


-----Original Message-----
From: Confocal Microscopy List <[hidden email]> On Behalf Of Wendy Salmon
Sent: 15 February 2021 21:40
To: [hidden email]
Subject: Looking for recipe to create an Andor FRAPPA calibration slide?

This email was sent to you by someone outside the University.
You should only click on links or attachments if you are certain that the email is genuine and the content is safe.

*****
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.
*****

Hi all,
Before I shell out cash to buy one, has anyone reverse-engineered the Andor FRAPPA calibration slide and would be willing to share?  It's the smear of Ethidium Bromide doped agarose (not the front surface mirror for the MicroPoint system).
Thank you!
Wishing you peace and good health,
Wendy

--------------------
Wendy C Salmon, M.A. (she/her/hers)
Manager, W.M. Keck Facility for Biological Imaging

Whitehead Institute for Biomedical Research
455 Main St., Rm 447, Cambridge, MA 02142 USA
e: [hidden email]  |  w: microscopy.wi.mit.edu

<http://microscopy.wi.mit.edu>
The University of Edinburgh is a charitable body, registered in Scotland, with registration number SC005336. Is e buidheann carthannais a th’ ann an Oilthigh Dhùn Èideann, clàraichte an Alba, àireamh clàraidh SC005336.

[Gary Laevsky]

*****
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.
*****

That's right, I forgot!

We used, and still use, fluorescently labeled agarose.

Also, Zazzle (others probably work) marker on a coverslip works well.

We also use the Zazzle marker for TIRF demo. Dot the coverslip, and then
add water.  Some of the Zazze juice sticks, and the rest forms
mini-micelles.  Works great, easy, and cheap!

On Wed, Feb 24, 2021 at 7:49 AM WHEELER Ann <[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.
> *****
>
> Hi Wendy
>
> Yes, Well sort of. I used fluorescently labelled fibronectin and it did a
> very similar job. Although it was ages ago, I have a mosaic on my Andor
> system now not a FRAPPA. Essentially you need a fluorescent thing you can
> make a hole in and then calibrate the xy galvos of the FRAPPA to the
> position of the hole if my memory serves me correctly. I'm sure
> flurosecently labelled agarose would do the same job. The Fluorescently
> labelled Fibronectin can be brought from Thermo for about £350.
>
> Hope this helps
>
> Ann
>
> Advanced Imaging Resource, Institute of Genetics and Molecular Medicine,
> University of Edinburgh, Edinburgh EH4 2XU
> Working Pattern. Mon 9am - 2pm, Tues - Thurs 9am - 5pm.
> E: [hidden email]
> T: 0131 651 8665
> W: http://www.ed.ac.uk/igmm-imaging
>
>
> -----Original Message-----
> From: Confocal Microscopy List <[hidden email]> On
> Behalf Of Wendy Salmon
> Sent: 15 February 2021 21:40
> To: [hidden email]
> Subject: Looking for recipe to create an Andor FRAPPA calibration slide?
>
> This email was sent to you by someone outside the University.
> You should only click on links or attachments if you are certain that the
> email is genuine and the content is safe.
>
> *****
> 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.
> *****
>
> Hi all,
> Before I shell out cash to buy one, has anyone reverse-engineered the
> Andor FRAPPA calibration slide and would be willing to share?  It's the
> smear of Ethidium Bromide doped agarose (not the front surface mirror for
> the MicroPoint system).
> Thank you!
> Wishing you peace and good health,
> Wendy
>
> --------------------
> Wendy C Salmon, M.A. (she/her/hers)
> Manager, W.M. Keck Facility for Biological Imaging
>
> Whitehead Institute for Biomedical Research
> 455 Main St., Rm 447, Cambridge, MA 02142 USA
> e: [hidden email]  |  w: microscopy.wi.mit.edu
>
> <http://microscopy.wi.mit.edu>
> The University of Edinburgh is a charitable body, registered in Scotland,
> with registration number SC005336. Is e buidheann carthannais a th’ ann an
> Oilthigh Dhùn Èideann, clàraichte an Alba, àireamh clàraidh SC005336.
>

--
Best,

Gary Laevsky, Ph.D.
Director, Confocal Imaging Facility
Nikon Center of Excellence
Co-Founder, North Atlantic Microscopy Society (NAMS)
https://namsmicroscopy.com/
Dept. of Molecular Biology
Washington Rd.
Princeton University
Princeton, New Jersey, 08544-1014
(O) 609 258 5432
(C) 508 507 1310