Re: Phase Contrast problems-our experience

[Asson-Batres, Mary Ann]

I am resending this message because I received an error message indicating it could not be sent because it duplicated a similar message.  Please forgive the redundancy if you get the same message twice, but if not, please teach me what I am doing wrong.  Thank you....

 

Mary Ann

 

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From: Asson-Batres, Mary
Sent: Wednesday, September 24, 2008 8:15 AM
To: Confocal Microscopy List
Subject: RE: Phase Contrast problems

We have had a similar problem when viewing sections.  Try removing the plate and you will likely see the abberant image even without a specimen in view.  It turned out for us that this was due to condensation on the lens of the CCD camera.  We had to have a technician clean the lens and reseal it.  No problems since then. 

Mary Ann

Mary Ann Asson-Batres

Department of Biological Sciences

Tennessee State University

3500 John A Merritt Blvd

Nashville, TN  37209

 

Office  615-963-5779

FAX    [phone]

 

________________________________________
From: Confocal Microscopy List [[hidden email]] On Behalf Of Nowell, Cameron [[hidden email]]
Sent: Wednesday, September 24, 2008 7:45 AM
To: [hidden email]
Subject: Re: Phase Contrast problems

Hi Matt,
            We have the same microscope and do not have any problems imaging phase through culture plates. Phase works fine on plastic, DIC does not usually work well (or at all).

You say you have aligned the microscope for phase, does this include Kohler Illumination? What camera are you using? Can you see the same effect down the eye peices of the scope?


Cheers


Cam


Cameron J Nowell
Microscopy Research and Imaging Core Facility
Peter MacCallum Cancer Centre
12 St Andrews Place
East Mebourne, 3002
Victoria, Australia

Phone: [phone]
Fax:       [phone]
Mobil:  [phone]


________________________________

From: Confocal Microscopy List on behalf of Matthew Pearson
Sent: Wed 24/09/2008 9:16 PM
To: [hidden email]
Subject: Phase Contrast problems



Hi all,

I have a problem when viewing 6-well plates using a Leica DMIRB invert microscope and imaging using phase contrast.  When I go to the live camera feed in our acquisition software I can see what looks like a reflection or the image of the condenser annulus in the image.  It looks as though the light is passing through the annulus and then projects its image onto the CCD.  This doesn't happen when imaging slides and only happens sometimes with 6-well plates.  The plates are plastic, is phase contrast possible through plastic or does it have to be glass?  I have aligned the microscope for phase contrast imaging using the Bertrand lens, but this does not appear to solve the problem.  I am using a 5x Ph0 objective and the corresponding Ph0 condenser annulus.

I would appreciate any thoughts on this.

thanks,

Matt Pearson.







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