Micro stonemasons. Difflugia.

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c-krebs
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Micro stonemasons. Difflugia.

#1 Post by c-krebs » Tue Apr 28, 2020 6:45 am

When one of my marsh samples has run it's course, I sometimes check through the muck and mud at the bottom to see what wonderful little sculptures reside there. The testate amoebae Difflugia are quite common, but I still enjoy studying them. And it is hard to resist taking a few more images of them.

Olympus 20/0.40 LMPLFLN, Canon 90D.
Image

Olympus 50/0.50 LMPLN, Canon 90D.
Image

Nikon LU Plan 50/0.80. Canon 90D
Image

Nikon LU Plan 50/0.80. Canon 90D
Image

Wes
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Re: Micro stonemasons. Difflugia.

#2 Post by Wes » Tue Apr 28, 2020 6:49 am

Amazing images, what technique did you use?

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75RR
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Re: Micro stonemasons. Difflugia.

#3 Post by 75RR » Tue Apr 28, 2020 6:49 am

Very nice! Are they building in quartz?

Wonder if that is a deliberate choice (translucent house) or just available material - and yes I know they have no 'brain' :)
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DonSchaeffer
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Re: Micro stonemasons. Difflugia.

#4 Post by DonSchaeffer » Tue Apr 28, 2020 10:14 am

Who needs a brain when you can do this. Wonderful photos. The look like Grecian Urns.

Bruce Taylor
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Re: Micro stonemasons. Difflugia.

#5 Post by Bruce Taylor » Tue Apr 28, 2020 10:56 am

75RR wrote:
Tue Apr 28, 2020 6:49 am
Very nice! Are they building in quartz?

Wonder if that is a deliberate choice (translucent house) or just available material - and yes I know they have no 'brain' :)
A French group did a rather ingenious study of Difflugia's material-selection. I wrote a blog entry about it a few years ago: https://www.itcamefromthepond.com/tag/difflugia/

Short version: yes, they choose their building materials, and quartz is preferred. :)

Stunning images, Charlie!

(On the subject of "building beautiful things without brains," I sometimes remind myself that cell's in a chicken's skin have the ability to create wonderfully intricate structures called "feathers". :D Which is to say, the wondrousness of cellular life is everywhere you look!).

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KD Arvidsson
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Re: Micro stonemasons. Difflugia.

#6 Post by KD Arvidsson » Tue Apr 28, 2020 11:10 am

Very amazing! //KD
Microscope Nikon Labophot 2
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https://www.youtube.com/channel/UCjsgbq ... dyl2x0Atpw

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Re: Micro stonemasons. Difflugia.

#7 Post by Chris Dee » Tue Apr 28, 2020 11:14 am

Great images and subject treatment. Are you happy with the Canon 90D? I'm considering buying one, any downsides?

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Re: Micro stonemasons. Difflugia.

#8 Post by Hobbyst46 » Tue Apr 28, 2020 12:56 pm

Extremely beautiful images!
Please, some more details about the technique: illumination, stacking, post-processing... thanks in advance!
And a question: given that the structures are quartz - are they colored under polarized light ?
Zeiss Standard GFL+Canon EOS-M10, Olympus VMZ stereo

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Re: Micro stonemasons. Difflugia.

#9 Post by RobBerdan » Tue Apr 28, 2020 5:49 pm

Very nice photos.
RB

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c-krebs
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Re: Micro stonemasons. Difflugia.

#10 Post by c-krebs » Wed Apr 29, 2020 1:39 am

Bruce...
Thanks for the link to your informative article. The video of Difflugia baccillariarum was great. I really wish I came across them more often.
Hobbyst46...
Please, some more details about the technique: illumination, stacking, post-processing... thanks in advance!
And a question: given that the structures are quartz - are they colored under polarized light ?
Here a small picture of how the two last ones were illuminated:
Image
The microscope is a Nikon MM-11 set up with an Olympus Super Widefield (infinity) head. I use a wide variety of objectives on it. It is set up to do "direct projection" onto an APS-C body on the trinocular tube. This particular 50X has a pretty good NA (0.80) but a very small working distance. Often I'll use a translucent hemisphere with a hole in it for the objective, but with this very small working distance light can only get in from the sides, so a ring of Lee (or Rosco) diffusion material is sufficient and it is positioned around the subject. The tiny quartz particles would give very strong specular highlights without some diffusion, which would make stacking very problematic I move the lights (continuous LED) around and adjust them until I like what I see. I have outfitted the fine-focus knob with a 400 step stepper motor which is controlled with a WeMacro controller. I stack with both Helicon and Zerene. Helicon is much faster, but it's retouching ability is not as good. Often I will tag my stacks and load them into Helicon (batch more) and run a variety of methods and settings. Then I study the results decide if any are worth pursuing to a finished image. I make a determination as to what program and method to use for the final image. I'll do what "retouching" can be done in the stacking programs, and then pull it into Photoshop to complete it.

A few years back I posted some other testate amoebae and one was a "quartz" Difflugia under crossed polarizers. Yes they are colored. The post is here:
viewtopic.php?f=6&t=3624&p=32844#p32844

Chris...
I got the 90D because I want a higher resolution APS-C; a fully electronic shutter from live-view; and I wanted a camera that could do a quick focus stack sequence when used outdoors or for lower magnification close-ups. It does all three. Like all other cameras it is not "perfect", but the only real thing I'm a little uncertain about is the noise level. It's got 32Mp, which I wanted for direct projection with 5/10/20X objectives. It is far more than you would need for most microscope set-ups. At first blush, when you look at 1:1, the noise level is higher than I was used to from APS-C with lower pixel counts. The primary time it can get annoying is when you run a very long stack in either PMax (Zerene) or "C" in Helicon. It does clean up quite nicely in a program like Neat Image.

Some people think it is a weird hybrid between a DSLR and a "Mirrrorless", but I actually like the configuration. (Still not quite ready to give up an optical viewfinder when outdoors). I will say that if your microscope configuration is suitable for a full frame camera (and you have a collection of Canon "glass") you might want to look at the EOS R. I've been using that lately on my BHS with the 2.5X NFK, and have been very pleased.

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Re: Micro stonemasons. Difflugia.

#11 Post by PeteM » Wed Apr 29, 2020 5:21 am

Spectacular images.

Thanks, also, for explaining a bit about how you took them.

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Re: Micro stonemasons. Difflugia.

#12 Post by Chris Dee » Wed Apr 29, 2020 11:51 am

c-krebs wrote:
Wed Apr 29, 2020 1:39 am
Chris...
I got the 90D because I want a higher resolution APS-C; a fully electronic shutter from live-view; and I wanted a camera that could do a quick focus stack sequence when used outdoors or for lower magnification close-ups. It does all three. Like all other cameras it is not "perfect", but the only real thing I'm a little uncertain about is the noise level. It's got 32Mp, which I wanted for direct projection with 5/10/20X objectives. It is far more than you would need for most microscope set-ups. At first blush, when you look at 1:1, the noise level is higher than I was used to from APS-C with lower pixel counts. The primary time it can get annoying is when you run a very long stack in either PMax (Zerene) or "C" in Helicon. It does clean up quite nicely in a program like Neat Image.

Some people think it is a weird hybrid between a DSLR and a "Mirrrorless", but I actually like the configuration. (Still not quite ready to give up an optical viewfinder when outdoors). I will say that if your microscope configuration is suitable for a full frame camera (and you have a collection of Canon "glass") you might want to look at the EOS R. I've been using that lately on my BHS with the 2.5X NFK, and have been very pleased.
Many thanks for the rundown on the 90D Charles. While looking at EOS R reviews I learned Canon is stopping development for the EF mount, and that they've hamstrung EF-S lenses on the EOS R via own adaptor (30 fps limit and 11MP effective crop). I'm going to have a rethink of my options but your reply was helpful and appreciated.

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Re: Micro stonemasons. Difflugia.

#13 Post by Hobbyst46 » Wed Apr 29, 2020 1:11 pm

Thanks, Charles, for the details.
Zeiss Standard GFL+Canon EOS-M10, Olympus VMZ stereo

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Re: Micro stonemasons. Difflugia.

#14 Post by daruosha » Wed Apr 29, 2020 2:08 pm

That's a very sophisticated setup and the results speak for themselves. Thanks for sharing.
Daruosh.

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Re: Micro stonemasons. Difflugia.

#15 Post by Scarodactyl » Thu Apr 30, 2020 2:43 am

I had never heard of these--what a delightful microbe! The photos are stunning too.

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Re: Micro stonemasons. Difflugia.

#16 Post by MikeBradley » Wed May 06, 2020 6:27 am

I read with interest the reasons Charles gave for selecting the 90D. Does it suffer from the vibration issues that plagued the 60D and were still evident to a lesser degree in to 70D and 80D models despite them having EFCS?
Thanks
michael
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c-krebs
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Re: Micro stonemasons. Difflugia.

#17 Post by c-krebs » Thu May 07, 2020 4:53 am

MikeBradley wrote:
Wed May 06, 2020 6:27 am
I read with interest the reasons Charles gave for selecting the 90D. Does it suffer from the vibration issues that plagued the 60D and were still evident to a lesser degree in to 70D and 80D models despite them having EFCS?
When used as a DSLR (not in "live-view") it behaves pretty much the same as the other xxD Canons.
When you go to "live-view" it becomes nearly identical to the EOS M6II. It really feels like a mirrorless camera. In live-view you can set three shutter options:
1- Mechanical
2- Electronic First Shutter Curtain
3_Electronic

The second, EFSC, still has that annoying little vibration and sound that all xxDs have had since the 50D. (I'd put it the sound about the same as the 80D, the slight vibration maybe a little less).
But the third choice, "Electronic", is fully electronic. It is absolutely silent with zero vibration. (If you use it with a lens attached, and the lens is not at full aperture you will hear slight sound as the aperture closes and reopens. But on a microscope... nothing!). Its the only way I use live-view with continuous light, and it is great.

There is no flash signal to hotshoe or built in flash when using electronic shutter. (Generally the case with most cameras. Certain Olympus models will fire flash with fully electronic shutter but the sync speed is only 1/20 sec).
If set for EFSC, a (Canon dedicated) flash will fire but at the moment of exposure it switches to the mechanical shutter. Since I don't like the initial double-shutter action of that sequence: "shutter down-shutter open-flash-shutter down-shutter open", I'll often switch back to SLR mode with mirror lock-up. I release the camera in anticipation of a shot, and the mirror goes up. Then when I eventually release the shutter it is a single shutter action (opens) before the flash.

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Re: Micro stonemasons. Difflugia.

#18 Post by MikeBradley » Thu May 07, 2020 7:06 am

This is very helpful Charles, I couldn't get this info from the manual.
One other question concerns the remote operation, is it possible to trigger the shutter using either their IR remote or the hardware switch when tethered to a computer running EOS Utility or some such program via usb cable?
Thanks
Michael
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Carl Zeiss Standard WL
Canon 90D

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Re: Micro stonemasons. Difflugia.

#19 Post by c-krebs » Thu May 07, 2020 7:48 am

Michael,
I was afraid you might ask about that... I'm just not the tethering type! ;)

There's no IR remote. And I have no idea about a second method or triggering when connected to EOS Utilities.

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Re: Micro stonemasons. Difflugia.

#20 Post by ImperatorRex » Thu May 07, 2020 8:12 am

c-krebs wrote:
Thu May 07, 2020 4:53 am
...If set for EFSC, a (Canon dedicated) flash will fire but at the moment of exposure it switches to the mechanical shutter. Since I don't like the initial double-shutter action of that sequence: "shutter down-shutter open-flash-shutter down-shutter open", I'll often switch back to SLR mode with mirror lock-up. I release the camera in anticipation of a shot, and the mirror goes up. Then when I eventually release the shutter it is a single shutter action (opens) before the flash.
Charles,
what is your experience, does the SLR modes (thats avoids the initial double-shutter action) provide actual benefit or better results? For example if the organism reaactes sensible on vibrations? I also using the SLR mode but it is maybe more a psychological thing and preference.

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Re: Micro stonemasons. Difflugia.

#21 Post by c-krebs » Thu May 07, 2020 6:57 pm

ImperatorRex wrote:
Thu May 07, 2020 8:12 am
Charles,
what is your experience, does the SLR modes (thats avoids the initial double-shutter action) provide actual benefit or better results? For example if the organism reaactes sensible on vibrations? I also using the SLR mode but it is maybe more a psychological thing and preference.
I suppose there might be a "psychological" element in there (i.e. thinking one shutter movement must have less vibration, and be quicker than two). But with my older cameras I used the DSLR/mirror-up mode for flash because the delay time (from triggering to flash) seemed noticeably shorter than triggering from live-view. It's the method I've used for years, so I suppose I just continued working that way with the 90D. Certainly the "mirror-up" vibration can spook some creatures. But generally when I use flash I'm chasing some little subject around the slide, or trying to stop motion of some sort. So the shortest delay is very desirable.

I don;t have the proper instrumentation at home to test delay times and vibration, but perhaps I should just do some subjective testing. My impression is that the mirror and shutter mechanism on the 90D are better in this respect compared to my older cameras, but I have nothing empirical to back that up.

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Re: Micro stonemasons. Difflugia.

#22 Post by ImperatorRex » Thu May 07, 2020 7:52 pm

Thank you very much for the detailed explaination Charles. Never thought on the delay time and if the difference can be recognized. But delay time may not be relevant anyway if the photo is done with the Canon EOS Utility "Life-View mode". The mirrow it locked open for many of the EOS modells, if I remember correctly.

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