I would like the hear from those who have used DIC (Nomarski) microscopy to examine minute anatomy of preserved, stained micro-invertebrates, specifically rotifers. I have been told that DIC is not the best technique for stained specimens. Any opinions on that?
My opinion is that it works just fine. Specimens do not have to be stained to observe them with DIC, but if that is the material you have at hand and if it si important to record details that are difficult to make out without it, then I think this DIC is the way to go. The staining will affect the image, no doubt about that, but unless you are using a birefringent dye (anything forming crystals can be problematic), this won't make it imposible to get a good image of what you want to show. I have seen award winning images of stained specimens obtained with DIC. And I think they had scientific value as well.
I agree with Miloš. DIC is very fine to observe rotifers whatever if they are stained or not. DIC is usefull to highlight transparent structures that could also be visible with some stainings. But the use DIC microscopy with stained specimens can be problematic if the tranparency of those specimens is lost due to an exceed of stain.
As a consequence, instead of asking if DIC is a good method to observe stained rotifers, I would wonder if it is usefull to stain them when you can use DIC microscopy.
I agree with Milos and Frederic. I have found DIC has proved excellent for anatomical details of the mites I work with (most in the range of 200-500 microns in length). But there are a couple of points not raised by Milos and Frederic where DIC might not prove so good: (1) it works best on flat or nearly-flat specimens. Oval or spherical objects do not provide the same level of sharp contrast; (2) there is some variation in the quality of Normarski optical systems between different brands of compound microscopes. For example the system used by Leitz in the 1990s (e.g. on the Leitz Laborlux model) produced images that were inferior to Olympus and Zeiss.
DIC will work with stained speciments well, if they are not havily overstained. It is used for example to check the IN SITU stainning of minute creatures.
Aydin,
I have used DIC extensively for more than 30 years with numerous stained invertebrates, including most phyla. It can work beautifully, including with rotifers. I have published many of these images in books and publications. I've added a link below to a book I produced that uses DIC extensively with stained inverts across most phyla.
Bruce
Book Atlas of Invertebrate Reproduction and Development, Second Edition
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