MicrobeHunter.com Microscopy Forum

Another diatom from the muddy water of the pothole in the yard. I intended to take a focus stack, but it was moving all the time so I did the best I could with a single shot.

1. Bright field, 20x objective:


2. DIC, 20x objective:

To me these look very good for one shot each.
CE

Gekko,

I agree with CE. Very nice. You have mastered the art of the single exposure. I like both photos and especially the DIC, even though it may give some false relief. It makes the lipid globules much more apparent. Three cheers for a job well done.

lorez

CE and lorez, thank you very much for your kind (actually too kind) comments. I must admit that I've not had good results with focus stacks even when the diatoms were stationary. True the DIC image does highlight the lipid globules but I think I should have toned down the DIC effect somewhat. Thanks again.

Bonjour Gekko
Belles séries images
Cordialement seb

seb, thank you again for your kind comment.

I like the last one, which almost look like a school book illustration. Looks like a painting almost. Nice effect. Is that achieved in the post processing or a "DIC effect"? I have not experienced such effect on my DIC setup (yet)

hkv wrote:I like the last one, which almost look like a school book illustration. Looks like a painting almost. Nice effect. Is that achieved in the post processing or a "DIC effect"? I have not experienced such effect on my DIC setup (yet)

Thank you, khv: I very much appreciate any feedback. I think I have overdone the DIC (set the prism for maximum effect) which gives it an exaggerated (fake) 3D effect. But I'm not sure what you are referring to exactly. On my microscope, the DIC prism incorporates a retardation plate, so as I move the slider, the color changes. I usually try to set it for a neutral gray background, but in the image captured by the camera, the background comes out as a deep cyan; I don't know why. In editing, I found that if I use the eyedropper on the background to render it gray, the colors of the diatom (or whatever the object is) also change accordingly (as can be expected). On the other hand, if I desaturate the cyan background color only, then only the background changes to gray, the colors of the object remain unchanged (I guess unless the object contains cyan color), so that is what I did here. After that, I adjusted levels, contrast, resized the image for posting, and sharpened it. But the first step is: in camera raw (I use Adobe Elements) I adjust the exposure slightly if necessary, then reduce the noise, then open the image in the editor where I may reduce noise further if necessary. I sometimes attempt to (partially or completely) clean the background when there is "junk" or out-of-focus blobs, depending on whether I think it is distracting or I leave it in if I think it may give "context" (or if I am too lazy). Sometimes the image sort of looks like a water color painting if I use high ISO and then overdo the noise reduction, but in this case I used a low ISO (200) so I don't think that could explain what you describe. Perhaps I oversharpened it? I don't really know. To me the image looks rather "harsh" which I attribute to the strong DIC setting I used. Does any of this explain what you see? Thanks again.
One last possibility: "empty magnification". If I view the image smaller, it looks better to my eyes. I've reduced the size and posted again below: does that help? Thanks again.