Re: Coverslip

Posted by Shalin Mehta on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Coverslip-tp592448p592451.html

Dear Monique,

We need to use confocal and collect reflected laser. One will need to setup a light path that allows passage of reflected laser to a detector which has pin-hole in front - which should be possible on most of the microscopes as the dichroic never completely gets rid of excitation laser. At air-glass and glass-air interfaces, laser will be reflected much more strongly than within coverslip. This residual laser from dichroic should be detectable if the emission filter is removed from the path. The confocal pin-hole will provide strong depth descrimination and allow measurement of coverslip to submicron accuracy.

The correction factor that I mentioned assumes normal incidence of laser on coverslip, so the use 'beam-parking' or 'point-scanning' mode at the center of field of view is required.

Regards
Shalin

On Mon, Feb 25, 2008 at 11:12 PM, Vasseur Monique <[hidden email]> wrote:

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How to do you that? See it?  What is the microscope setup?

 

Monique Vasseur

Microscopie et imagerie

Département de biochimie

Université de Montréal

C.P. 6128, succursale Centre-ville

Montréal QC    H3C 3J7   Canada

tél. [phone] poste 5148

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De : Confocal Microscopy List [mailto:[hidden email]] De la part de Rietdorf, Jens
Envoyé : 24 février 2008 15:30
À : [hidden email]
Objet : Re: Coverslip

 

Dear Sarah,

 

you could use reflections from the coverglass which are characteristic for upper and lower edge of the cover glass and easy to spot.

 

regards, jens

 

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Dr. Jens Rietdorf[hidden email]
Head Microscopy
Novartis Research Foundation
Friedrich-Miescher-Institute, wro1066.2.16 Maulbeerstr.66, CH-4058 Basel, Switzerland
phone [phone] mobil [phone]
Email:rietdorf(at)fmi.ch

 

 

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From: Confocal Microscopy List [mailto:[hidden email]] On Behalf Of Sarah Kefayati
Sent: Freitag, 22. Februar 2008 21:42
To: [hidden email]
Subject: Coverslip

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

Hello All:

 

I need to know how I can measure the depth from the cover slip,is there any marked or fluorescent coverslip that I can use for this purpose?

 

Thanks for any suggestion

Sarah

--
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Shalin Mehta
mobile: [phone]
blog: shalin.wordpress.com
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Bioimaging Lab, Block-E3A, #7-10                
Div of Bioengineering, NUS Singapore 117574  
website: http://www.bioeng.nus.edu.sg/optbioimaging/colin/index.html

Liver Cancer Functional Genomics Lab, #6-05
National Cancer Centre, Singapore 169610
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