December 14, 2017, 05:30:50 AM

Author Topic: 5Dsr MP-E 65 z-stacking experiments.  (Read 16380 times)

NancyP

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #15 on: July 27, 2015, 11:49:12 AM »
Thanks for info, Zeidora

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #15 on: July 27, 2015, 11:49:12 AM »

Boromir883

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #16 on: August 11, 2015, 04:15:07 AM »
Thanks Zeidora. It`s impressiv how many details show up, when equipment is used at its max  :)

Zeidora

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #17 on: August 11, 2015, 11:07:28 AM »
Thanks. I agree, seeing individual cells with "simple" SLR is cool. But there are limits to imaging SLR. To see cell surface sculpture I have to use the scanning electron microscope. Haven't done compound microscope imaging with epi-objectives, but most cell surface detail is too small for that anyway. And I like running the SEM :-)
5D2 full spectrum, 5DsR, a bunch of Zeiss (some other) primes, for documentary natural history, macro, and micro.

Boromir883

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #18 on: August 11, 2015, 11:57:13 AM »
Thanks. I agree, seeing individual cells with "simple" SLR is cool. But there are limits to imaging SLR. To see cell surface sculpture I have to use the scanning electron microscope. Haven't done compound microscope imaging with epi-objectives, but most cell surface detail is too small for that anyway. And I like running the SEM :-)
I would like to have access to such a toy  ;D

Zeidora

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #19 on: August 11, 2015, 08:51:34 PM »
Thanks. I agree, seeing individual cells with "simple" SLR is cool. But there are limits to imaging SLR. To see cell surface sculpture I have to use the scanning electron microscope. Haven't done compound microscope imaging with epi-objectives, but most cell surface detail is too small for that anyway. And I like running the SEM :-)
I would like to have access to such a toy  ;D
Yep, one of those job perks. Not compainin' about that one!
5D2 full spectrum, 5DsR, a bunch of Zeiss (some other) primes, for documentary natural history, macro, and micro.

Mt Spokane Photography

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #20 on: August 12, 2015, 12:37:37 AM »
I guess we will have to see what Neuro can produce with his conformal microscope

Zeidora

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #21 on: August 12, 2015, 01:32:46 AM »
I guess we will have to see what Neuro can produce with his conformal microscope
Ah, dam you autocorrect! ;-) Confocal works with laser excited fluorescent markers, generally of translucent objects. I am unaware of epic-confocal microscopy. I guess you could extract the uppermost layer with computer processing of images, but that obviates the entire point of confocal.
Both SLR imaging and SEM are for surface structures, so completely different imaging approaches. For SLR, the object is generally completely untreated and un-stained. For SEM the specimen is often pre-treated (e.g., critical point drying of flowers), sometimes fixed/stained (e.g. with nasty osmium tetroxide, haven't done that in a long time), and generally also sputter coated with precious metals, mostly gold and/or palladium. I work quite a bit with uncoated material in variable pressure.
5D2 full spectrum, 5DsR, a bunch of Zeiss (some other) primes, for documentary natural history, macro, and micro.

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #21 on: August 12, 2015, 01:32:46 AM »

Boromir883

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #22 on: August 12, 2015, 01:51:45 AM »
Zeidora,
-as a person with a degree in biotechnology (but working as quality Manager in food industry) i like reading about your working technics  :)

Zeidora

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #23 on: August 12, 2015, 02:28:56 AM »
Zeidora,
-as a person with a degree in biotechnology (but working as quality Manager in food industry) i like reading about your working technics  :)
Thanks! Just in case you're interested, I wrote a couple of articles on imaging small flowers and z-stacking, and also a book chapter on scientific photography of mollusks. If interested, PM me and I send you links.
5D2 full spectrum, 5DsR, a bunch of Zeiss (some other) primes, for documentary natural history, macro, and micro.

malus

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #24 on: November 17, 2017, 04:21:56 PM »
sorry for reopening a old post.
I am considering using the mp-e to take high resolution images of small (0.1 to 1 mm) crystals and I am wondering if Moiere interference can be an issue with the 5dsr. Does anyone have experience with that? Is the 5ds a better option?


Zeidora

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #25 on: November 17, 2017, 04:34:10 PM »
sorry for reopening a old post.
I am considering using the mp-e to take high resolution images of small (0.1 to 1 mm) crystals and I am wondering if Moiere interference can be an issue with the 5dsr. Does anyone have experience with that? Is the 5ds a better option?
If you want significantly <1 mm full frame where every pixel contains information (not glorified part of a blur circle), you want to go rather on a compound microscope with high NA epi objectives. Consider that on 5DsR with MP-E65 at f/2.8 maximum mag where you get info out on 5DsR is 4:1. Even at 5:1 you just get blur circles. 4:1 means field of view is 4x6 mm, so a 0.5 mm structure will be rather small. If the pixel dimensions are sufficient for your purposes, then that's fine, but otherwise go compound. If that is something you are interested, reply and I will provide details.

Hope that helps.
5D2 full spectrum, 5DsR, a bunch of Zeiss (some other) primes, for documentary natural history, macro, and micro.

malus

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #26 on: November 18, 2017, 06:23:49 AM »
I should have mentioned that I already have access to all sort of microscopic equipment: from magnifier lenses to state of the art TEMs.
 For this project I need some space to access the crystals during the shots. Unfortunately compound microscopes do not have enough working distance for my need. The stereomicroscope I currently use has very bad glasses and ccd. A new one with some decent specs cost much more than a camera, which I could also use for other purposes.
I agree that it is a waste of money/time if I can't get some decent pictures.

Zeidora

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #27 on: November 18, 2017, 11:15:57 AM »
I should have mentioned that I already have access to all sort of microscopic equipment: from magnifier lenses to state of the art TEMs.
 For this project I need some space to access the crystals during the shots. Unfortunately compound microscopes do not have enough working distance for my need. The stereomicroscope I currently use has very bad glasses and ccd. A new one with some decent specs cost much more than a camera, which I could also use for other purposes.
I agree that it is a waste of money/time if I can't get some decent pictures.

How much do you need? is the problem bulk of specimen and limited stage travel, or are your specimens very long needles? There are long WD objectives, usually with a bit lower NA. Alternatively, use compound lens on photographic equipment. If you have older 160 mm tube length lenses, then use them on bellows. If you have more modern ICS objectives, then you need a relay lens. Then specimen bulk is of no concern.

If you also have access to VP or E SEM and chamber is sufficiently large, then you can go un-coated in VP or E, use long WD to get as much DOF as possible, and shoot at high probe current. 0.1 mm is pretty low mag on SEM, so you could go 500-1000 pA. You can even z-stack SEM images. Haven't done that myself, but know of people who have done that.
5D2 full spectrum, 5DsR, a bunch of Zeiss (some other) primes, for documentary natural history, macro, and micro.

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #27 on: November 18, 2017, 11:15:57 AM »

zim

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #28 on: November 18, 2017, 11:45:34 AM »
Definitely a thread which deserves to bubble back to the surface, inspiring stuff, fantastic when such technical info is shared too.

@Zeidora are you still using AP to finish off your images?

malus

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #29 on: November 18, 2017, 12:21:24 PM »
@zeidora: I seem to understand form your comments that you are proper microscopist but I don't want to bother other readers with technical stuff.
I am interested in monitoring crystals during solid-state reactions. In some case these occur by varying temperature, in other cases by exposing crystals to humidity, pressure, or different gasses including HCl and ammonia, through the use of special cells that I have designed. Now for some reason people don't want me to use HCl in their ESEM  :-X.
Being more serious most of these reactions produce a change of the crystal colour that electron microscopy cannot reveal.

Anyway if as you shouw above the lens resolution is the limiting factor then the test results of DoX mark suggest that I'm better off with a 6D or 5D and an extender than the 5DS(r).

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Re: 5Dsr MP-E 65 z-stacking experiments.
« Reply #29 on: November 18, 2017, 12:21:24 PM »