Posted by
David Claypool on
URL: http://confocal-microscopy-list.275.s1.nabble.com/DIC-condensers-tp7583008p7583052.html
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Non-commercial response:
Jeff,
In my experience, use a Planapochromat 60/1.4 or 100/1.45 oil objective, 1.4NA oil condenser, precision interference line green filter(546/10), and "standard" or "high" contrast DIC objectives & DIC condenser prisms. You will need to carefully test which combination of DIC prisms work best for you. Be sure the back aperture of the objective is completely filled. You will probably need to introduce a 1.5x-2.5x magnification factor in the camera light path, and, in this case, a solid state illuminator should give you more light than a 100W halogen lamp. Some microscope systems offer a high NA illuminator adapter that will accept a solid state illuminator liquid light guide, and this works well for this application. You will probably need to adjust your image histogram for the best image.
I recently used the above combination on an inverted microscope and achieved excellent results with a 5MP sCMOS camera.
David
David J. Claypool
Digital Imaging Product Manager
Micro Video Instruments
Office: 800-875-2041 x5221
Cell: 603-809-5342
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> On Nov 28, 2014, at 9:30 PM, "Jeff Spector" <
[hidden email]> wrote:
>
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>
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>
> Greetings,
> This isn't exactly a "confocal" questions but I know a lot of "micoscopy
> gurus" live on this list so I thought it a good place to ask this. I have a
> colleague who is trying to image individual (i.e. small and diffraction
> limited) microtubules in a flow chamber by using DIC. They are currently
> using a 100x 1.45 oil Objective, but only a .52 LWD condenser. They were
> using a solid state light source but couldn't get good image so we switched
> to a lamp for illumination and the images are much better, and we can now
> see the microtbules but there still isn't a lot of contrast. My question
> is, is it worth it to go to a high NA (perhaps oil immersion) condenser,
> and can anyone think of why the lamp would give a better DIC image than a
> solid state light source?
> thanks in advance for the help...
> -Jeff