Calibration/Grid slide for use with fluroescence?

[Adrian Smith-6]

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Hi all,

Does anyone know of a source of calibration/grid/micrometer slides that will work with fluorescence rather than transmitted light? Ideally, it would work with widefield, laser scanning confocal and multiphoton.

I've heard (third-hand) of using an etched stage micrometer and "painting" a fluorescent solution on to it that will sit in the etchings but I've not found someone who has actually done it... (and I would prefer not to have to deal with liquids each time I want to use it).

Any suggestions gratefully received.

Regards,

Adrian


______________________________________________
Dr Adrian Smith
    Manager, Cytometry & Imaging Facilities      
    Centenary Institute
    http://www.centenary.org.au     
    Locked Bag No.6 Newtown, NSW 2042 AUSTRALIA.                                                                    
    Ph: 61-2-9565-6189     Fax: [phone]

[Cameron Nowell]

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Hi Adrian,

I just use a broken haemocytometer i have lying around. It is pretty
auto fluorescent in visible light so works well for both transmitted and
widefield fluorescence. I have not however tried it on a confocal. Might
give it a go and see what happens.


Cheers

Cam



Cameron J. Nowell
Microscopy Manager
Centre for Advanced Microscopy
Ludwig Institute for Cancer Research
PO Box 2008
Royal Melbourne Hospital
Victoria, 3050
AUSTRALIA
Office: [phone]
Mobile: [phone]
Fax: [phone]
Facility Website



-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]]
On Behalf Of Adrian Smith
Sent: Monday, 16 May 2011 5:35 PM
To: [hidden email]
Subject: Calibration/Grid slide for use with fluroescence?

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Hi all,

Does anyone know of a source of calibration/grid/micrometer slides that
will work with fluorescence rather than transmitted light? Ideally, it
would work with widefield, laser scanning confocal and multiphoton.

I've heard (third-hand) of using an etched stage micrometer and
"painting" a fluorescent solution on to it that will sit in the etchings
but I've not found someone who has actually done it... (and I would
prefer not to have to deal with liquids each time I want to use it).

Any suggestions gratefully received.

Regards,

Adrian


______________________________________________
Dr Adrian Smith
    Manager, Cytometry & Imaging Facilities      
    Centenary Institute
    http://www.centenary.org.au     
    Locked Bag No.6 Newtown, NSW 2042 AUSTRALIA.

    Ph: 61-2-9565-6189     Fax: [phone]

[Mark Cannell]

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If you calibrate the scanner in transmitted mode, it will be  
calibrated for all other modes.

Hope this helps,

Mark


On 16/05/2011, at 7:35 PM, Adrian Smith wrote:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all,
>
> Does anyone know of a source of calibration/grid/micrometer slides  
> that will work with fluorescence rather than transmitted light?  
> Ideally, it would work with widefield, laser scanning confocal and  
> multiphoton.
>
> I've heard (third-hand) of using an etched stage micrometer and  
> "painting" a fluorescent solution on to it that will sit in the  
> etchings but I've not found someone who has actually done it... (and  
> I would prefer not to have to deal with liquids each time I want to  
> use it).
>
> Any suggestions gratefully received.
>
> Regards,
>
> Adrian
>
>
> ______________________________________________
> Dr Adrian Smith
>    Manager, Cytometry & Imaging Facilities
>    Centenary Institute
>    http://www.centenary.org.au
>    Locked Bag No.6 Newtown, NSW 2042 AUSTRALIA.
>    Ph: 61-2-9565-6189     Fax: [phone]

[Katharina Thomsen]

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Hi Adrian,

you may ask ibidi (http://www.ibidi.de/). They had developed a
possibility to write structures with an laser, also in fluorescence
standards. Probably you don't find it on the website, you just contact them.

Regards
Katharina

no commercial interest

Am 16.05.11 09:35, schrieb Adrian Smith:

> *****
> To join, leave or search the confocal microscopy listserv, go to:
> http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
> *****
>
> Hi all,
>
> Does anyone know of a source of calibration/grid/micrometer slides that will work with fluorescence rather than transmitted light? Ideally, it would work with widefield, laser scanning confocal and multiphoton.
>
> I've heard (third-hand) of using an etched stage micrometer and "painting" a fluorescent solution on to it that will sit in the etchings but I've not found someone who has actually done it... (and I would prefer not to have to deal with liquids each time I want to use it).
>
> Any suggestions gratefully received.
>
> Regards,
>
> Adrian
>
>
> ______________________________________________
> Dr Adrian Smith
>      Manager, Cytometry&  Imaging Facilities
>      Centenary Institute
>      http://www.centenary.org.au
>      Locked Bag No.6 Newtown, NSW 2042 AUSTRALIA.
>      Ph: 61-2-9565-6189     Fax: [phone]

[Keith Morris]

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Dear Adrian,

For graticule and eyepiece reticules, eg stage micrometer slides with
calibration scale bars, try

http://www.2spi.com/catalog/ltmic/stage-micrometers-graticules.php

http://www.graticules.com

SPi do supply cheaper calibration slides, although the superior Graticules
Ltd £300 stainless steel metal cased stage micrometer slide really do last
forever. It is nice to have a 100um scale [2um per division] and a 1mm scale
[10um per division] slide, for low and high mag objectives respectively,
although in these days of austerity I make do with just the 1mm scale I
acquired years ago that was checked against my old departments NPL certified
Graticules Ltd stage micrometer. Unless you work to full QA/GLP with
independent QA auditing you will not need £1000 certification by NPL [UK
National physics laboratory] or NIST [US].

You can easily capture a usable image of the stage micrometer scale under
fluorescence, because as the scale is a bit metallic it reflects as well as
blocks light [both under confocal and with widefield microscopes]. Just up
the laser power or the camera exposure time. The image of the calibration
scale under fluorescence is simply inverted [scale white, background black],
and the scale will be more than clear enough for calibration. Although as
Mark says the scale will be the same size as viewed by transmission light
[DIC/Phase Contrast off], as it's more about optical magnification than
light mode, but it's nice to see that they are identical.

For an image of a 1mm calibration scale by transmitted and fluorescence
light viewed using a 10x air objective and a 100x oil objective see:

http://www.well.ox.ac.uk/cytogenetics/calibrationscale10x.jpg

http://www.well.ox.ac.uk/cytogenetics/calibrationscale100x.jpg

It is 10um per division, so 0 to 10 will be 100um. The image was captured
using an Olympus BX-51 widefield upright microscope, although it looks
pretty identical scanned with our Zeiss 510 confocal. You just replace the
specimen slide with the stage micrometer graticule slide and capture an
image with each objective. Then use imageJ or MetaMorph (or Imaris or
ImageProPlus or Photoshop) to work out the um per pixel (or pixels per
micrometer) calibration factor for each objective. Easy to do as you know
the length of the photographed um scale in both um and pixels.

The only time you might need a different stage micrometer calibration slide
is with metallurgical microscopes that are used with samples without cover
slips (graticules.com make a special stage micrometer slide for this
application, and you don't need or want this type of calibration slide for
standard microscope imaging using slides with cover slips):

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:  [phone]
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 Adrian Smith
Sent: 16 May 2011 08:35
To: [hidden email]
Subject: Calibration/Grid slide for use with fluroescence?

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

Hi all,

Does anyone know of a source of calibration/grid/micrometer slides that will
work with fluorescence rather than transmitted light? Ideally, it would work
with widefield, laser scanning confocal and multiphoton.

I've heard (third-hand) of using an etched stage micrometer and "painting" a
fluorescent solution on to it that will sit in the etchings but I've not
found someone who has actually done it... (and I would prefer not to have to
deal with liquids each time I want to use it).

Any suggestions gratefully received.

Regards,

Adrian


______________________________________________
Dr Adrian Smith
    Manager, Cytometry & Imaging Facilities      
    Centenary Institute
    http://www.centenary.org.au     
    Locked Bag No.6 Newtown, NSW 2042 AUSTRALIA.

    Ph: 61-2-9565-6189     Fax: [phone]

[Adrian Smith-6]

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http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
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On 16/05/2011, at 5:35 PM, Adrian Smith wrote:
>
> Hi all,
>
> Does anyone know of a source of calibration/grid/micrometer slides that will work with fluorescence rather than transmitted light? Ideally, it would work with widefield, laser scanning confocal and multiphoton.


Thanks to everyone who made suggestions on this front.

I think I have fluorescence and confocal covered but I'm still not sure about multiphoton... particularly when said multiphoton is a dedicated system with no visible light lasers...

Regards,

Adrian

[Clements, Ian]

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Please join us for a free educational webinar!

"Live Cell Imaging Made Easier"

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During this informative webinar, we'll discuss:
* Key methods to improve live cell imaging
* Helpful hints for better experiment design
* The importance of the light path and the benefits of TruLight(tm)

For more information or to register, go to http://apiwebinars.webex.com


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Product Manager - DeltaVision OMX Systems


This email message, together with any attachments, is for the sole use of the intended recipient(s) and is the confidential information of Applied Precision Inc. If you are not the intended recipient, your review, use, disclosure, copying or dissemination of this email message or its attachments, or the information contained therein, is strictly prohibited. If you are not the intended recipient or if you think this email was sent to you in error, please notify the sender by reply email and delete this message and its attachments, as well as all copies, from your system.

[Barbara Foster]

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Hey, Ian

What's this I hear about API being acquired by GE?

Best regards,
Barbara
O: [phone]
C: [phone]

Bending the curve toward faster sales and business development


At 02:35 PM 6/2/2011, you wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Please join us for a free educational webinar!
>
>"Live Cell Imaging Made Easier"
>
>Presented by Dr. Katie Buchanan, DeltaVision Product Manager at
>Applied Precision
>
>Wednesday, June 29 at  6 a.m., 11 a.m. and 6 p.m. PST
>
>During this informative webinar, we'll discuss:
>* Key methods to improve live cell imaging
>* Helpful hints for better experiment design
>* The importance of the light path and the benefits of TruLight(tm)
>
>For more information or to register, go to http://apiwebinars.webex.com
>
>
>Ian Clements
>Product Manager - DeltaVision OMX Systems
>
>
>This email message, together with any attachments, is for the sole
>use of the intended recipient(s) and is the confidential information
>of Applied Precision Inc. If you are not the intended recipient,
>your review, use, disclosure, copying or dissemination of this email
>message or its attachments, or the information contained therein, is
>strictly prohibited. If you are not the intended recipient or if you
>think this email was sent to you in error, please notify the sender
>by reply email and delete this message and its attachments, as well
>as all copies, from your system.

[Robin Battye-2]

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It is posted on their website:

http://www.api.com/news/2011/2011.04.28.asp




-----Original Message-----
From: Confocal Microscopy List [mailto:[hidden email]] On
Behalf Of Barbara Foster
Sent: Friday, June 17, 2011 10:53 AM
To: [hidden email]
Subject: Re: Free Vendor Webinar - Live Cell Imaging Made Easier

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Hey, Ian

What's this I hear about API being acquired by GE?

Best regards,
Barbara
O: [phone]
C: [phone]

Bending the curve toward faster sales and business development


At 02:35 PM 6/2/2011, you wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Please join us for a free educational webinar!
>
>"Live Cell Imaging Made Easier"
>
>Presented by Dr. Katie Buchanan, DeltaVision Product Manager at Applied
>Precision
>
>Wednesday, June 29 at  6 a.m., 11 a.m. and 6 p.m. PST
>
>During this informative webinar, we'll discuss:
>* Key methods to improve live cell imaging
>* Helpful hints for better experiment design
>* The importance of the light path and the benefits of TruLight(tm)
>
>For more information or to register, go to http://apiwebinars.webex.com
>
>
>Ian Clements
>Product Manager - DeltaVision OMX Systems
>
>
>This email message, together with any attachments, is for the sole use
>of the intended recipient(s) and is the confidential information of
>Applied Precision Inc. If you are not the intended recipient, your
>review, use, disclosure, copying or dissemination of this email message
>or its attachments, or the information contained therein, is strictly
>prohibited. If you are not the intended recipient or if you think this
>email was sent to you in error, please notify the sender by reply email
>and delete this message and its attachments, as well as all copies,
>from your system.

[Barbara Foster]

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Sorry folks... just didn't check who was on the "reply" line!

Congrats are in order, apparently, for our colleagues at API.

Best regards,
Barbara Foster, President and Sr. Consultant

Microscopy/Microscopy Education
P: [phone]
W: www.MicroscopyEducation.com


At 10:27 AM 6/17/2011, you wrote:

>*****
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>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Hey, Ian
>
>What's this I hear about API being acquired by GE?
>
>Best regards,
>Barbara
>O: [phone]
>C: [phone]
>
>:35 PM 6/2/2011, you wrote:
>>*****
>>To join, leave or search the confocal microscopy listserv, go to:
>>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>>*****
>>
>>Please join us for a free educational webinar!
>>
>>"Live Cell Imaging Made Easier"
>>
>>Presented by Dr. Katie Buchanan, DeltaVision Product Manager at
>>Applied Precision
>>
>>Wednesday, June 29 at  6 a.m., 11 a.m. and 6 p.m. PST
>>
>>During this informative webinar, we'll discuss:
>>* Key methods to improve live cell imaging
>>* Helpful hints for better experiment design
>>* The importance of the light path and the benefits of TruLight(tm)
>>
>>For more information or to register, go to http://apiwebinars.webex.com
>>
>>
>>Ian Clements
>>Product Manager - DeltaVision OMX Systems
>>
>>
>>This email message, together with any attachments, is for the sole
>>use of the intended recipient(s) and is the confidential
>>information of Applied Precision Inc. If you are not the intended
>>recipient, your review, use, disclosure, copying or dissemination
>>of this email message or its attachments, or the information
>>contained therein, is strictly prohibited. If you are not the
>>intended recipient or if you think this email was sent to you in
>>error, please notify the sender by reply email and delete this
>>message and its attachments, as well as all copies, from your system.