A little guidance would be wonderful.

Have questions about the equipment used for macro- or micro- photography? Post those questions in this forum.

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Crumble
Posts: 6
Joined: Fri Mar 12, 2021 9:52 pm
Location: San Diego

A little guidance would be wonderful.

Post by Crumble »

Hey guys,

New to the forum, but have been in love with the microscopic world my whole life (both parents were microbioligists).

I own 2 microscopes, a Zeiss Universal with a set of Phase Neofluar objectives, that I use for Phase and Dark Field microscopy, and an AO Series 4 with a set of Bright Phase objectives for negative phase.

For my camera, I own a Canon EOS-M with Magic Lantern, and in the next few days will be ordering a Panasonic GH5 (I want to do video and still microscope imagery).

What I am looking for guidance on, is the best way to capture images and video with the trinocular ports of my microscopes. At the moment I am using a T2 to 0.5x relay lens tube to 23mm adapter, and have been disappointed with the sharpness in both images and video. Image is much sharper through the eyepieces. Also, I am unable to get the camera parafocal to the eyepieces.

I have been reading up on the forum, and am seeing some people using a bellow to hold the camera, and projection lens straight onto the sensor. Also seeing some mention of using a bellow, but with a 40mm lens (if APS-C sensor), with a normal 10x eyepiece for use with glasses?

If I wanted the absolute best image quality and sharpness, which would be the best way to go?

Also, another question... how would I hook up a bellow to the Universal, with how large it is?

Sorry for all the questions.

Thanks in advance for any insight!

Colin

Ichthyophthirius
Posts: 1152
Joined: Thu Mar 07, 2013 5:24 am

Re: A little guidance would be wonderful.

Post by Ichthyophthirius »

Crumble wrote:
Sat Mar 13, 2021 9:55 pm
I own 2 microscopes, a Zeiss Universal with a set of Phase Neofluar objectives

is the best way to capture images and video with the trinocular ports of my microscopes.
Hi Colin,

The Zeiss optics require compensating eyepieces, so the only way to adapt to the Universal is the "afocal" method. https://en.wikipedia.org/wiki/Afocal_photography

This means mounting a Zeiss Kpl eyepiece in a black or grey monotube https://www.mikroskopie-forum.de/index. ... #msg156073 https://www.mikroskopie-forum.de/index. ... #msg156088 on the trinocular port. Then use one of the Zeiss clamp adapters to mount your lens + camera above the eyepiece. You may need a dovetail-to-filter thread adapter that is rare and may have to be custom-made (Raf Camera may have some in stock).

This is the same optical arrangement as the human eye. The camera "looks" through the eyepiece with the lens set to infinity and an image is formed at the sensor.

As the eyepiece, use a high eyepoint Zeiss Kpl (glasses symbol) which are the best-corrected. For the camera lens, use a manual focus prime lens, ideally a short or even pancake lens. A low f-number like f2.8 is sufficient.

For APS-C, the combination I use is a 10x eyepiece and a 40 mm camera lens. This gives good coverage of an FN18 field of view. Don't bother with bellows. Other combinations, with 4x, 6.3x, and 8x are possible but 10x is straight-forward.
The best Zeiss 10x eyepieces are in this order (from very good to excellent and from affordable to expensive: Kpl-W 10x/18 < Kpl-W 10x/20 < Kpl-S 10x/20
As the camera lens you should get a Pentax-M SMC 40 mm f2.8 https://www.pentaxforums.com/lensreview ... -Lens.html You can also use 45 mm or 50 mm lenses but your field of view will be smaller. I vastly prefer manual lenses for this because of the mechanical stability and because they have a hard stop at infinity. The camera lens needs to be free of dust, as internal dust shows up badly in the images when using the afocal method.

Make sure your EOS-M has a vibration-free shutter (EFSC or electronic shutter) and will take images without a lens attached (as the manual lenses won't have the required contacts).

For m4/3 sensors I have no personal experience. When using a 10x eyepiece, the camera lens should have a focal length of around 30 mm. Here is the Sigma 30 mm http://www.savazzi.net/photography/zeissadapter.html though other sources report that some copies of the Sigma 30 mm have problems with onion ring artifacts. Maybe others here have suggestions for m4/3 sensors? Or maybe combine a Kpl 8x/18 with a 40 mm camera lens.

Regards, Ichty

Ichthyophthirius
Posts: 1152
Joined: Thu Mar 07, 2013 5:24 am

Re: A little guidance would be wonderful.

Post by Ichthyophthirius »

Crumble wrote:
Sat Mar 13, 2021 9:55 pm
If I wanted the absolute best image quality and sharpness, which would be the best way to go?
The Neofluar objectives you have aren't plan lenses. So you can only use the center of the image anyway. Many afocal adaptations will give you excellent center sharpness and contrast, even the simpler and cheaper ones. The very best solutions will further improve only the image corners, not the center. So you don't have to be too concerned with the edges; they are limited by the Neofluar design anyway.

Pau
Site Admin
Posts: 6051
Joined: Wed Jan 20, 2010 8:57 am
Location: Valencia, Spain

Re: A little guidance would be wonderful.

Post by Pau »

I agree with Ichthyophthirius.
Some more info on afocal:
viewtopic.php?p=99265#p99265
Pau

Crumble
Posts: 6
Joined: Fri Mar 12, 2021 9:52 pm
Location: San Diego

Re: A little guidance would be wonderful.

Post by Crumble »

Ichthyophthirius wrote:
Sun Mar 14, 2021 9:30 am
Crumble wrote:
Sat Mar 13, 2021 9:55 pm
I own 2 microscopes, a Zeiss Universal with a set of Phase Neofluar objectives

is the best way to capture images and video with the trinocular ports of my microscopes.
Hi Colin,

The Zeiss optics require compensating eyepieces, so the only way to adapt to the Universal is the "afocal" method. https://en.wikipedia.org/wiki/Afocal_photography

This means mounting a Zeiss Kpl eyepiece in a black or grey monotube https://www.mikroskopie-forum.de/index. ... #msg156073 https://www.mikroskopie-forum.de/index. ... #msg156088 on the trinocular port. Then use one of the Zeiss clamp adapters to mount your lens + camera above the eyepiece. You may need a dovetail-to-filter thread adapter that is rare and may have to be custom-made (Raf Camera may have some in stock).

This is the same optical arrangement as the human eye. The camera "looks" through the eyepiece with the lens set to infinity and an image is formed at the sensor.

As the eyepiece, use a high eyepoint Zeiss Kpl (glasses symbol) which are the best-corrected. For the camera lens, use a manual focus prime lens, ideally a short or even pancake lens. A low f-number like f2.8 is sufficient.

For APS-C, the combination I use is a 10x eyepiece and a 40 mm camera lens. This gives good coverage of an FN18 field of view. Don't bother with bellows. Other combinations, with 4x, 6.3x, and 8x are possible but 10x is straight-forward.
The best Zeiss 10x eyepieces are in this order (from very good to excellent and from affordable to expensive: Kpl-W 10x/18 < Kpl-W 10x/20 < Kpl-S 10x/20
As the camera lens you should get a Pentax-M SMC 40 mm f2.8 https://www.pentaxforums.com/lensreview ... -Lens.html You can also use 45 mm or 50 mm lenses but your field of view will be smaller. I vastly prefer manual lenses for this because of the mechanical stability and because they have a hard stop at infinity. The camera lens needs to be free of dust, as internal dust shows up badly in the images when using the afocal method.

Make sure your EOS-M has a vibration-free shutter (EFSC or electronic shutter) and will take images without a lens attached (as the manual lenses won't have the required contacts).

For m4/3 sensors I have no personal experience. When using a 10x eyepiece, the camera lens should have a focal length of around 30 mm. Here is the Sigma 30 mm http://www.savazzi.net/photography/zeissadapter.html though other sources report that some copies of the Sigma 30 mm have problems with onion ring artifacts. Maybe others here have suggestions for m4/3 sensors? Or maybe combine a Kpl 8x/18 with a 40 mm camera lens.

Regards, Ichty
Awesome, thanks so much for the info!

I have the grey monotube (47 30 20) for the Zeiss, I need to find that clamp adapter. Looks like the clamp adapters just clamp onto the outside of the monotube? Then I adjust their position for being parafocal? Not as hard of a setup as I was expecting, thankfully. Now that I know that my setup was not ideal, not sure if I should upgrade my Camera or not.

Ichthyophthirius
Posts: 1152
Joined: Thu Mar 07, 2013 5:24 am

Re: A little guidance would be wonderful.

Post by Ichthyophthirius »

Crumble wrote:
Sun Mar 14, 2021 10:27 pm
I have the grey monotube (47 30 20) for the Zeiss, I need to find that clamp adapter. Looks like the clamp adapters just clamp onto the outside of the monotube? Then I adjust their position for being parafocal? Not as hard of a setup as I was expecting, thankfully. Now that I know that my setup was not ideal, not sure if I should upgrade my Camera or not.
Hi Colin,

You don't need to change parfocality on the phototube. The grey tube "47 30 20" already has exactly the right length to make up a 160 mm optical tube length. If you put in a fixed (no diopter) Kpl 10x eyepiece, the tube length will be correct. Once the phototube is set up, it's the reference for the tube length. You achieve parfocality by changing the settings of the diopters on the binocular tube or the eyepieces in the binoculars not the other way around!

If you find one of the Zeiss bell-shaped adapters, you will also need a dovetail adapter to connect the bell and the filter thread of the lens. This one from RafCaerma will work: https://rafcamera.com/catalog/product/v ... mm-to-t2m/ plus you need a step down ring to connect the filter thread. Here you can see the principle: https://www.mikroskopie-forum.de/index. ... #msg196488 Make sure you get a 10x Kpl eyepiece without a diopter so the eyepiece fits inside the dovetail adapter.

There are other, cheaper adapters available as well like this Pentax microscope adapter https://www.ebay.com/itm/Asahi-Pentax-M ... 6456797490 (I deliberately linked to an expensive offer; you can get it for half the price). It should fit and you need am M42/T2 converter and a step down ring (like 49-42mm) to attach the camera lens. Not as nice or stable as the Zeiss adapter but it should work.

Put a sheet of paper around the monotube so the adapter won't damage the paint.

If you then set the camera lens to infinity, changing the position of the camera won't affect parfocality. That's part of being an afocal setup. Changing the position of the lens will however affect vignetting. There is an ideal position and distance between eyepiece and front lens of the camera (usually a few mm) that you have to find by trail and error. After that you don't change it any more.

---

I think the camera is rarely the issue when used on a microscope. All the digital EOS models should have enough image quality and resolution. It's however really important that they have EFSC or electronic shutter https://krebsmicro.com/Canon_EFSC/index.html (don't know if that is the case with your EOS-M). In the forum, we often find that vibrations are ruining images. They can come from the camera, or the table or the user. You can start tracking them down by doing a shutter speed series (5s, 1/2s, 1/20s, 1/200s) and comparing the results. Personally, I love Canon's EOS Utility software which lets you control your camera from a computer so I don't need to press the shutter on the camera body. I wouldn't get a camera without software either on a desktop or at least on a smartphone. If your EOS-M can't be controlled from your phone, do you have remote control for it?

It's also very common that the images out of the camera (even a very good one that is adapted correctly) don't look as good as what you see though the eyepieces. The human brain does a lot of "post-processing" :) You need to do a lot of optimisation of illumination and image processing to get to the level of image quality that your brain generates for free. Microscopy techniques like phase contrast or DIC that produce a grey background and strong edge contrasts make it easier to take good images - you're nicely set up with the excellent Neofluar phase set.

Regards, Ichty

Crumble
Posts: 6
Joined: Fri Mar 12, 2021 9:52 pm
Location: San Diego

Re: A little guidance would be wonderful.

Post by Crumble »

Ichthyophthirius wrote:
Mon Mar 15, 2021 3:10 am
Crumble wrote:
Sun Mar 14, 2021 10:27 pm
I have the grey monotube (47 30 20) for the Zeiss, I need to find that clamp adapter. Looks like the clamp adapters just clamp onto the outside of the monotube? Then I adjust their position for being parafocal? Not as hard of a setup as I was expecting, thankfully. Now that I know that my setup was not ideal, not sure if I should upgrade my Camera or not.
Hi Colin,

You don't need to change parfocality on the phototube. The grey tube "47 30 20" already has exactly the right length to make up a 160 mm optical tube length. If you put in a fixed (no diopter) Kpl 10x eyepiece, the tube length will be correct. Once the phototube is set up, it's the reference for the tube length. You achieve parfocality by changing the settings of the diopters on the binocular tube or the eyepieces in the binoculars not the other way around!

If you find one of the Zeiss bell-shaped adapters, you will also need a dovetail adapter to connect the bell and the filter thread of the lens. This one from RafCaerma will work: https://rafcamera.com/catalog/product/v ... mm-to-t2m/ plus you need a step down ring to connect the filter thread. Here you can see the principle: https://www.mikroskopie-forum.de/index. ... #msg196488 Make sure you get a 10x Kpl eyepiece without a diopter so the eyepiece fits inside the dovetail adapter.

There are other, cheaper adapters available as well like this Pentax microscope adapter https://www.ebay.com/itm/Asahi-Pentax-M ... 6456797490 (I deliberately linked to an expensive offer; you can get it for half the price). It should fit and you need am M42/T2 converter and a step down ring (like 49-42mm) to attach the camera lens. Not as nice or stable as the Zeiss adapter but it should work.

Put a sheet of paper around the monotube so the adapter won't damage the paint.

If you then set the camera lens to infinity, changing the position of the camera won't affect parfocality. That's part of being an afocal setup. Changing the position of the lens will however affect vignetting. There is an ideal position and distance between eyepiece and front lens of the camera (usually a few mm) that you have to find by trail and error. After that you don't change it any more.

---

I think the camera is rarely the issue when used on a microscope. All the digital EOS models should have enough image quality and resolution. It's however really important that they have EFSC or electronic shutter https://krebsmicro.com/Canon_EFSC/index.html (don't know if that is the case with your EOS-M). In the forum, we often find that vibrations are ruining images. They can come from the camera, or the table or the user. You can start tracking them down by doing a shutter speed series (5s, 1/2s, 1/20s, 1/200s) and comparing the results. Personally, I love Canon's EOS Utility software which lets you control your camera from a computer so I don't need to press the shutter on the camera body. I wouldn't get a camera without software either on a desktop or at least on a smartphone. If your EOS-M can't be controlled from your phone, do you have remote control for it?

It's also very common that the images out of the camera (even a very good one that is adapted correctly) don't look as good as what you see though the eyepieces. The human brain does a lot of "post-processing" :) You need to do a lot of optimisation of illumination and image processing to get to the level of image quality that your brain generates for free. Microscopy techniques like phase contrast or DIC that produce a grey background and strong edge contrasts make it easier to take good images - you're nicely set up with the excellent Neofluar phase set.

Regards, Ichty
Thanks so much!

I just ordered the RAF adapter, hopefully the 2-3 month delivery time is a conservative estimate... :shock:

Also ordered this, which looks to be the Zeiss bell adapter that I will need, plus it has a dovetail to 27mm filter ring adapter. https://www.ebay.com/itm/Microscope-ada ... SwS4dfuQ9J

I wonder if I could use that dovetail to 27mm adapter instead of the RAF adapter? Then just find a step up ring from 27mm to whatever size the filter is for my lens?

Yeah the EOS-M has electronic first shutter. So it must be using a relay lens instead of corrected eyepiece that is giving unsatisfactory sharpness. Below is an example of a focus stack with the current setup, ignore the poor white balance... :lol:

Thanks so much for all of the help!
Attachments
BrownTardScaled.jpg

Pau
Site Admin
Posts: 6051
Joined: Wed Jan 20, 2010 8:57 am
Location: Valencia, Spain

Re: A little guidance would be wonderful.

Post by Pau »

Yes, this is the right "bell" adapter
Crumble wrote:
Mon Mar 15, 2021 9:45 am
I wonder if I could use that dovetail to 27mm adapter instead of the RAF adapter? Then just find a step up ring from 27mm to whatever size the filter is for my lens?
very likely you will not have enough clearance for the eyepiece, did you see my third setup? viewtopic.php?p=224647#p224647
Yeah the EOS-M has electronic first shutter. So it must be using a relay lens instead of corrected eyepiece that is giving unsatisfactory sharpness.
I don't understand the last sentence. You need to use the right corrective eyepiece: KPL or S-KPL
It's better to use it at its right position in afocal mode, you can also use it raised as projective, but in both cases you need its compensation.
Pau

Crumble
Posts: 6
Joined: Fri Mar 12, 2021 9:52 pm
Location: San Diego

Re: A little guidance would be wonderful.

Post by Crumble »

Pau wrote:
Mon Mar 15, 2021 10:40 am
Yes, this is the right "bell" adapter
Crumble wrote:
Mon Mar 15, 2021 9:45 am
I wonder if I could use that dovetail to 27mm adapter instead of the RAF adapter? Then just find a step up ring from 27mm to whatever size the filter is for my lens?
very likely you will not have enough clearance for the eyepiece, did you see my third setup? viewtopic.php?p=224647#p224647
Yeah the EOS-M has electronic first shutter. So it must be using a relay lens instead of corrected eyepiece that is giving unsatisfactory sharpness.
I don't understand the last sentence. You need to use the right corrective eyepiece: KPL or S-KPL
It's better to use it at its right position in afocal mode, you can also use it raised as projective, but in both cases you need its compensation.
Oh, yes I didn't consider the eyepiece clearance, so looks like I'll have to wait for the RAF adapter to arrive.

Last sentence I was just talking to myself out loud, confirming that my relay lens was the main culprit. :lol: I have KPL eyepieces, with the glasses symbol. Sadly they have delamination, bought them cheap because I didn't think I would need them other than for observing. So will have to buy one more with no delamination.

Ichthyophthirius
Posts: 1152
Joined: Thu Mar 07, 2013 5:24 am

Re: A little guidance would be wonderful.

Post by Ichthyophthirius »

Yeah, the opening of a 27 mm thread is probably too small.

Still a good find for the bell adapter. Theoretically, you could use the black cylinder but it's not easy. You would have to drill several holes in a filter ring adapter, then use the screw holes on top of the cylinder (the ones that presently hold the bajonet mount) to attach the filter ring. I wouldn't just glue it to the top as that's too dangerous for the camera.

It'll be more stable with the Raf adapter.

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