Stage micrometers and diatoms

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Shalin Mehta Shalin Mehta
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Stage micrometers and diatoms

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear confocalists,

 Is there a stage micrometer that works in both transmission and reflection mode- for measuring true magnification? I have diatom slide from Mr. Klaus Kemp with 8 diatoms that allow evaluation of resolution. Are some features on diatom frustule known to have certain distance between them? Following diatoms are mounted on the slide.

  • Amphipleura pellucida
  • Frustulia rhomboides
  • Pleurosigma angulatum
  • Surirella gemma
  • Nitzschia sigma
  • Stauroneis phoenicentron
  • Navicula lyra
  • Gyrosigma balticum
Best,
Shalin
--
My co-ordinates:
Shalin Mehta, Graduate student
Graduate Programme in Bioengineering, NUS, Singapore
Email: shalin {dot} mehta {at} gmail {dot} com
Blog: electricsbm.blogspot.com
Mobile: +65 90694182
Gary Laevsky-2 Gary Laevsky-2
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Re: Stage micrometers and diatoms

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

We used to use chrome on glass micrometers from Applied Precision for just this purpose.  Works great!

 

Best,

 

Gary Laevsky, Ph.D.

Imaging Application Specialist

 

Andor Technology

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From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Shalin Mehta
Sent: Friday, August 10, 2007 8:38 AM
To: [hidden email]
Subject: Stage micrometers and diatoms

 

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear confocalists,

 Is there a stage micrometer that works in both transmission and reflection mode- for measuring true magnification? I have diatom slide from Mr. Klaus Kemp with 8 diatoms that allow evaluation of resolution. Are some features on diatom frustule known to have certain distance between them? Following diatoms are mounted on the slide.

  • Amphipleura pellucida
  • Frustulia rhomboides
  • Pleurosigma angulatum
  • Surirella gemma
  • Nitzschia sigma
  • Stauroneis phoenicentron
  • Navicula lyra
  • Gyrosigma balticum

Best,
Shalin
--
My co-ordinates:
Shalin Mehta, Graduate student
Graduate Programme in Bioengineering, NUS, Singapore
Email: shalin {dot} mehta {at} gmail {dot} com
Blog: electricsbm.blogspot.com
Mobile: +65 90694182

Barbara Foster Barbara Foster
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Re: Stage micrometers and diatoms

In reply to this post by Shalin Mehta
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi, Shalin,

In answer to your various questions:
a. A transmitted light micrometer that has been made using chrome on glass will work in both transmitted and reflected light
b. Yes, there are features on diatoms that have specific spacing and Mr. Kemp most likely provided a reference sheet with your diatom slide.  Mine is currently waiting to be unpacked, but I am sure some of the diatomists in the audience can provide those numbers off the top of their heads.  This particular set of diatoms is was chosen to give a range of those spacings.  If you can't get an answer, I'll contact you when I get my bench unpacked.

Best regards,
Barbara



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At 07:37 AM 8/10/2007, you wrote:
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear confocalists,

 Is there a stage micrometer that works in both transmission and reflection mode- for measuring true magnification? I have diatom slide from Mr. Klaus Kemp with 8 diatoms that allow evaluation of resolution. Are some features on diatom frustule known to have certain distance between them? Following diatoms are mounted on the slide.

  • Amphipleura pellucida
  • Frustulia rhomboides
  • Pleurosigma angulatum
  • Surirella gemma
  • Nitzschia sigma
  • Stauroneis phoenicentron
  • Navicula lyra
  • Gyrosigma balticum
Best,
Shalin
--
My co-ordinates:
Shalin Mehta, Graduate student
Graduate Programme in Bioengineering, NUS, Singapore
Email: shalin {dot} mehta {at} gmail {dot} com
Blog: electricsbm.blogspot.com
Mobile: +65 90694182
Julio Vazquez Julio Vazquez
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Re: Stage micrometers and diatoms

In reply to this post by Shalin Mehta
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear diatom lovers, 

below are the spacings of the features of the different species on Klaus Kemp's 8-form test plate. I believe I got this info from the Carolina Biological supply web site, when they used to sell the slide.

There also used to be an article by Dave Walker: Counting teh dots: Giving Microscopes a workout using diatom test slides at  
www.microscopy-uk.org.uk/mag/artoct99/dwdiatom.html    

the article seems to be currently inaccessible, possibly being revised

Julio. 


  • Amphipleura pellucida 270 nm
  • Frustulia rhomboides 300 nm
  • Pleurosigma angulatum 525 nm
  • Surirella gemma 500 nm
  • Nitzschia sigma 440 nm
  • Stauroneis phoenicentron 720 nm
  • Navicula lyra 1250 nm
  • Gyrosigma balticum 670 nm



--
Julio Vazquez
Fred Hutchinson Cancer Research Center
Seattle, WA 98109-1024




On Aug 10, 2007, at 5:37 AM, Shalin Mehta wrote:

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear confocalists,

 Is there a stage micrometer that works in both transmission and reflection mode- for measuring true magnification? I have diatom slide from Mr. Klaus Kemp with 8 diatoms that allow evaluation of resolution. Are some features on diatom frustule known to have certain distance between them? Following diatoms are mounted on the slide.

  • Amphipleura pellucida
  • Frustulia rhomboides
  • Pleurosigma angulatum
  • Surirella gemma
  • Nitzschia sigma
  • Stauroneis phoenicentron
  • Navicula lyra
  • Gyrosigma balticum
Best,
Shalin
--
My co-ordinates:
Shalin Mehta, Graduate student
Graduate Programme in Bioengineering, NUS, Singapore
Email: shalin {dot} mehta {at} gmail {dot} com
Blog: electricsbm.blogspot.com
Mobile: +65 90694182

Julio Vazquez Julio Vazquez
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Re: Stage micrometers and diatoms

In reply to this post by Shalin Mehta
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
-

Shalin, 

When we purchased our Zeiss confocal, we got several test slides from Zeiss; these include a grid pattern of 2 micron wide (If I recall correctly) lines at 100 micron spacing, and a grating of 0.5 micron lines at 1 micron spacing (alternating dark and light 0.5 micron lines). In both cases, the lines are made of opaque/reflective thin metal deposit, I believe. These work well for measuring magnification and resolution/contrast. I am not sure where Zeiss gets them, or whether they sell them independently of their confocals, but if you have a contact for a Zeiss rep, might be worth asking them...


Another source of micrometers is Applied Image Inc...   they cost around $ 400.00. I think they call them "scales"




Carolina Biological Supply company has several conventional stage micrometers for about $ 30.00-50.00. Don't know if any of those work in reflected light...


Google "Carolina Biological" , and then search for micrometer

--
Julio Vazquez, 
Fred Hutchinson Cancer Research Center
Seattle, WA 98109-1024







On Aug 10, 2007, at 5:37 AM, Shalin Mehta wrote:

Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Dear confocalists,

 Is there a stage micrometer that works in both transmission and reflection mode- for measuring true magnification? I have diatom slide from Mr. Klaus Kemp with 8 diatoms that allow evaluation of resolution. Are some features on diatom frustule known to have certain distance between them? Following diatoms are mounted on the slide.

  • Amphipleura pellucida
  • Frustulia rhomboides
  • Pleurosigma angulatum
  • Surirella gemma
  • Nitzschia sigma
  • Stauroneis phoenicentron
  • Navicula lyra
  • Gyrosigma balticum
Best,
Shalin
--
My co-ordinates:
Shalin Mehta, Graduate student
Graduate Programme in Bioengineering, NUS, Singapore
Email: shalin {dot} mehta {at} gmail {dot} com
Blog: electricsbm.blogspot.com
Mobile: +65 90694182

Steffen Dietzel Steffen Dietzel
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Re: Stage micrometers and diatoms

In reply to this post by Shalin Mehta
Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

At 14:37 10.08.2007, you wrote:

>  Is there a stage micrometer that works in both
> transmission and reflection mode- for measuring true magnification?

Hi Shalin,

I used a USAF 1951 target for exactly that
purpose. It has a mirror surface with a clear
pattern on it. It appears there are several
vendors for those. I got it from Edmund
http://www.edmundoptics.com/

I believe the one that I used currently costs 111
Euros in Germany (cat. no. Y38-256). I switched
work places and just reordered it, so I can't
check the cat number on the slide itself.

One note of caution: I had some trouble with a
version of the slide that contains fluorescent
material. The focal plain is so far away from the
surface, that the working distance of my oil
immersion objectives was too short to reach it.
So you're better off with the one with the
surface mirror and use that in reflection or
transmission mode, as you plan anyway.

Steffen

--
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Ludwig-Maximilians-Universität München
Walter-Brendel-Zentrum (WBZ)
Head of light microscopy

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