Hello from Norfolk UK

[Pinguicula]

Hello there

I've always enjoyed Natural History Photography but since being semi retired I am able to explore macro and now micro photography in much more detail. Generally I'm using a Nikon D850 + Nikon 200m f4 and a 6T filter. Up to now I haven't used flash but the sun was just too much this year so I am beginning to explore a Nikon D500 + 105mm macro + Raynox 250 + Nikon Sb 700 with an AK diffuser.

I've use hand photostacking in the field for Bryophytes and Fungi and have just started with a 10x microscope objective. I've found Alan Wall's site invaluable to help get me started.

This is Greater Whipwort (Bazzania trilobata) a rather nice liverwort photographed in the depths of Wales. A stack of about 20 shots enabled full focus across the image. D850 +200mm f4 + 6T Filter

This is Feather Stalkball (Onygena corvina) growing on dead Razorbill. A stack of about 40-50 shots, D850 +200mm f4 + 6T Filter

A deligthful Earpick Fungus (Auriscalpium vulgare) A stack of about 30-40 shots, D850 +200mm f4 + 6T Filter

I love Slime Molds A stack of about 30 shots, D850 +200mm f4 + 6T Filter

These are my first attempts with the 10 x Objective Nikon D850 Nikon PD6 Bellows + Nikon 10 x Objective and Raynox 150(reversed) as the tube lens. I have a Stackshot and I am in the process of making a cage. Although these first two attempts were taken just in daylight. I'm using mirror up, 11µm steps and each one took between 120 - 180 shots.

This is Lachnella alboviolascens. But as you can see there's lots of problems with focus and even seeing through to the substrate under the fungus. This is what I would very like to learn how to avoid or at least how to ameliorate.

This is a small orange fungus (Nectria decora) that grows on dead twigs. In this case a Field Maple twig. As you can see the focus problem is again very obvious.

So I'm very much looking forward with your help and discussion to improve on this.

Kind regards & Happy New year

Mike

[JayMcClellan]

Hello from Michigan! Those are great shots, thanks for sharing. I just watched a good video from Alan Walls about stacking artifacts like you are seeing, and how to reduce or eliminate them. Maybe you would find it helpful if you haven't already seen it:
https://m.youtube.com/watch?v=uxxzZXORXL0

[Pinguicula]

Hello Jay

Thank you for the link. I hadn't discovered that one.

Thanks again

Mike

[Chris S.]

Mike,

Welcome to the forum!

I enjoyed your introductory images very much. And with most of them, I thought about my own travails lighting similar specimens.

I hope you keep posting in our community!

Very best regards,

--Chris S.

[Pinguicula]

Hello Chris

Thank you for the welcome and kind comments.

The low light capabilities of the D850 make the difference I think. All of these were taken in natural light including the two using the x10 objective.

Kind regards

Mike

Taken with a Nikon D850 + 200mm f4 + 6T Close up filter. Dusk fast approaching on a Graveyard Wall in Norfolk. About 25 shots. Mirror up mode using a remote release RFN-TX by SMDV

A Liverwort - Wall Scalewort (Porella platyphylla)

[ChrisR]

Really nce fingi, I don't think I've seen those .
I know this was a while ago - sorry I didn't see it.
I watched Allan Walls' video, which is all fine, but one thing he didn't mention:

When you use your nice wide aperture for resolution, the cone of light from the aperture to a point on the inside of that Lachnella alboviolascens fungus is wide, so the cone hits those light frilly bits, which at that slice, are out of focus, and give a light fuzz which overwhelms the dark detail you want.
(The half angle in air is arcsin(NA) )

If you stop down by say 3 stops, your cone of light is much narrower. It misses the frills. Great. You get much narrower halos.

Yes you get less resolution, and
yes you need 8x more light, and
yes you need to mess about, finding the best aperture and slice to retouch from.

How to stop a microscope objective down? Use a disc of black paper across the back with a round hole in it. When I did that I expected perspective changes to be a problem but it must have been telecentric or just lucky enough, because there wasn't a problem. It's actually years ago since I did it, but I'm pretty sure that when the alignment was close enough, Pmax + Zerene's alignments, with the best frames, picked the sharpest detail out just fine.
A much simpler proposition with a macro lens, especially in the field!

[rjlittlefield]

Pinguicula, sorry but I missed this one also!

This is Lachnella alboviolascens. But as you can see there's lots of problems with focus and even seeing through to the substrate under the fungus. This is what I would very like to learn how to avoid or at least how to ameliorate.

For reasons explained by ChrisR, "transparent foreground" is a serious problem at wide apertures typical of microscope objectives.

With simpler subjects, and using Zerene Stacker, one can use a targeted Stack Selected before retouching, as explained at https://zerenesystems.com/cms/stacker/d ... foreground .

However, for complex subjects a better approach is to use a "slabbing" workflow, then retouch from the slabs. There are many fewer slabs than original source images, so this is often a significant time saving. See https://zerenesystems.com/cms/stacker/docs/slabbing . Allan Walls is very fond of this technique.

This is a small orange fungus (Nectria decora) that grows on dead twigs. In this case a Field Maple twig. As you can see the focus problem is again very obvious.

This image has a different type of problem that is called "loss of detail halo". What you're showing looks typical of Zerene Stacker DMap when used with a contrast threshold slider that was set higher than it really should have been. When all or most of the frame is full of rich detail, it works better to set the slider low so as to keep as much of that detail as possible. However, when there is significant separation between foreground and background, the halo regions are also contaminated by out-of-focus foreground which is a separate but related problem. In those cases there is no perfect solution, but often the best result is obtained by rendering with both DMap and PMax and then combining the best bits with retouching. Again, Allan Walls is a fan of that technique.

--Rik