Fresh water crustacean larvae, bright, dark and DIC compared

Linden.g · Post by **Linden.g** » Mon Oct 03, 2011 8:34 pm

I often get asked to show DIC compared with bright field. I've also included dark field. 20x Achromat, focus stacks with flash.

Bright Field

DIC

Dark Field

pwnell · Post by **pwnell** » Mon Oct 03, 2011 9:00 pm

Aesthetically the DIC illumination is miles ahead.

Mitch640 · Post by **Mitch640** » Tue Oct 04, 2011 3:49 am

While they are all beautiful images of great quality, the brightfield one tells me more about the bug. That's how he really looks. Thanks for the extra effort. I didn't know you could go from one to the other that fast.

Pau · Post by **Pau** » Tue Oct 04, 2011 6:43 am

Mitch640 wrote:... the brightfield one tells me more about the bug. That's how he really looks. ...

Well, that's how it looks just under a brightfield microscope, but what really means?. Without microscope? (almost invisible)

Linden, excellent images and illustration of the illumination differences. Perhaps the brightfield one need to be more bright and contrasted to match the typical BF observation.

Mitch640 · Post by **Mitch640** » Tue Oct 04, 2011 6:49 am

Pau, all I can do is brightfield, so thats how they look to me. Without the microscope, you could never see them at all, bright, dark or DIC.

Pau · Post by **Pau** » Tue Oct 04, 2011 9:39 am

Mitch, yes, I understood your comment, but I found a good subject to post a little point for reflexion. For exemple, a natural looking photo isn't natural because all photos are artificial.
A photomicrography always need some manipulation of the subject and light. If it is free of artifacts (structures not present in the subject itself when alive) and it is both informative and artistically pleasent, you have a good one.

PD: If you can do BF, you also can do DF and oblique with a simple stop in the condenser.

Starshade · Post by **Starshade** » Tue Oct 04, 2011 10:21 am

Nice pics and cool object. I personally like the brighfield image among them.

However, had it been up to me which method to choose, I would stack both brightfield and DIC images to get maximum details (and do that semi-manually) and then get the legs out of DIC image and put them into brightfield and then fiddle around with the colours to make them match. In addition it's possible to consider merging DIC and brightfield of the middle parts, though it's harder.

May be it takes a load of manipulation but I would support those who think that photomicrographs are all about light and digital manipulations since we are unable to see the creatures themselves.

Bernd · Post by **Bernd** » Tue Oct 04, 2011 12:27 pm

Nice pics indeed. But remember: under the microscope, irrespective of the illumination used, none of the critters actually look "natural". Why? Everything looks flat due to the very limited depth of focus of the microscope objective. In real live all these critters always are 3-dimensional and more or less sphaerical.

Bernd

Linden.g · Post by **Linden.g** » Tue Oct 04, 2011 1:33 pm

Reprocessed, to brighten up bright field image

ChrisR · Post by **ChrisR** » Tue Oct 04, 2011 3:59 pm

Very interesting to see, thanks.
All three show structures you can't see in the other two.

But None of them IS a crustacean.
They're all representations of a crustacean, so none seems any more or less "valid" to me.

Wim van Egmond · Post by **Wim van Egmond** » Wed Oct 05, 2011 12:22 am

Very nice, and actually I prefer the darkfield image because it is more tranquile, the DIC image is a bit messy and the red eye is lost a bit.

But that is personal taste. In many case the DIC looks most spectacuar because it gives such a good view of the inside of a transparent organism. With the darkfield it is not possible to distinguish the features inside.

Mitch, you should try to create darkfield. It is not that difficult but you need a condenser with a higher NA than the NA of the objective you use. Start with low mag; 10X objective.

Wim