Advice please: High NA air objective with small WD

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Chris S.
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Advice please: High NA air objective with small WD

Post by Chris S. »

Seeking advice, please.

I've been offered an apo, high NA, air objective designed for no cover slip, that has a tiny working distance of 1.3mm. This objective has a wide front end that will make it nearly impossible to shine reflected light onto the subject. So in my view, subjects for this lens must either be thin and entirely backlit, or be illuminated by light shining through the lens. ("Axial illumination?" I'll use this term, but welcome correction if wrong.)

To use this lens on my current macro rig (the Bratcam), I would need to add a beamsplitter, light source from the side, and perhaps a polarization arrangement to stop light from bouncing off the interior surface of the objective. Pretty sure I could build this, but wish to do so only if time and cost prove worthwhile by permitting photographs that can not be obtained by simpler methods, and show uniquely informed insights.

I also have a Mitutoyo FS-70 head that I've been meaning to integrate into a working vertical rig. This head includes fiber optic axial illumination, which might or might not work well with this objective.

Can anyone with knowledge of axial illumination (if I'm using this term correctly) help me gain a sense of how much additional information, if any, this lens, plus system modifications needed to use it, would offer? Would any additional visual information be worth the trouble and cost? Axial illumination looks, to my uneducated eye, like a microscopic mug shot; are there ways to make this lighting more illustrative?

In advance, thanks. This lens is significantly discounted against list price for a new specimen; but the discounted price is still a lot of money--too much for me unless the lens lets me share significant new information in our forum.

Cheers,

--Chris S.

enricosavazzi
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Post by enricosavazzi »

I have no specific knowledge to offer, but I have a Mitutoyo FS-60 microscope equipped with four Mitutoyo M Plan Apos, and I rarely use the built-in axial illumination because it gives a very flat and very low contrast "rabbit-in-the-headlights" rendering of almost all subjects. It is especially hopeless on flat reflective subjects.

That said, a high-NA objective should be able to illuminate the subject with a broad cone of light converging onto the subject, while my M Plan Apos have at most an NA of 0.42 and therefore produce an illumination cone with a rather acute apex angle. This might make a difference, at least with some subjects.

The beamsplitter in the FS scopes is extremely thin, and likely so fragile that it is virtually impossible to clean, except perhaps by removing it and immersing it carefully into a cleaning solution.
--ES

TheLostVertex
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Re: Advice please: High NA air objective with small WD

Post by TheLostVertex »

Chris S. wrote:I've been offered an apo, high NA, air objective designed for no cover slip, that has a tiny working distance of 1.3mm. This objective has a wide front end that will make it nearly impossible to shine reflected light onto the subject.
I dont have any comments on on an axial solution, but I have used a 0.6mm wd objective for reflected light without too many issues. The shape of the objective might have been different though. Either way, is building some sort of coaxial ring light a possible option? I would guess that might give better images than a purely axial solution if you can manage.

Chris S.
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Post by Chris S. »

Enricco,

Thanks for sharing your experience with the FS-60 microscope and its built-in axial illumination. “Flat and very low contrast ‘rabbit-in-the-headlights’ rendering” is exactly what I was afraid of. I certainly don’t want it.

The NA on this objective is 0.90, so the illumination cone should have an obtuse angle (128 degrees?), but I don’t think I’ll risk the significant cost of this objective in hopes of using the wide entrance cone to create better modeling.

Steven,

Thanks for the suggestion of a co-axial ringlight. The front of this objective has a diameter of over 32mm, though—so in this case, the approach would create extreme side-lighting.

I’m going to pass on this objective. The $950 USD offer is a big discount compared with the new price of $6300, but is too much for the limited use I can envision for the thing. The lens in question is a Mitutoyo HR 100x/0.90.

Gentlemen, your experience and thoughts have been very helpful.

Cheers,

--Chris S.

Scarodactyl
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Post by Scarodactyl »

That's certainly a rare objective on the used market. I have found 1ish mm fine for oblique lighting with a fiber optic illuminator, though obviously not as easy as a more reasonable wd. That is at much lower mag though, with subjects that are more forgiving.

Macro_Cosmos
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Post by Macro_Cosmos »

I do have a beamsplitter cube coming which I intend to use for diffuse axial illumination.
Similar to transillumination/backlit, axial illumination/coaxial light works too. Personal preference thing I guess, or I may be wrong.

From my experience, I dislike this kind of setup a lot. The light lacks any kind of control, images all almost looks the same and flat. One would also lose a sizable percentage of light, 50% in my case. Open to change my views though, hope the incoming one does better than the ones I use at work.

I've also encountered bright hotspot issues due to the objective lens I use at work. It's not a microscope objective. The hotspot can be removed with polarisers, but images turn to mush, too much for that low MP camera.

I hope it works out in my favour for photography though, the morpho didius butterfly and wafers work incredibly well with epi-illumination.

I suggest cube beamsplitters, they are true beamsplitters.
Info here: https://www.edmundoptics.de/knowledge-c ... splitters/

You'll have 50% of light that comes out of one end which is wasted. You must have a way to deal with it. I use a flocked tube that's blocked off at the end.

Several forum users have seen success with reflected light for 1-2mm WD objectives. It will help if you can somehow get rid of the front housing.

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Post by rjlittlefield »

In principle, I think it should be possible to structure the illumination beam so as to provide asymmetric lighting for modeling, by sending light through only a part of the rear aperture. But I have not tried engineering such a beast, so I have no good idea how difficult that would be. (Murphy's Law predicts it's pretty hard.)

--Rik

Chris S.
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Post by Chris S. »

rjlittlefield wrote:In principle, I think it should be possible to structure the illumination beam so as to provide asymmetric lighting for modeling, by sending light through only a part of the rear aperture. But I have not tried engineering such a beast, so I have no good idea how difficult that would be. (Murphy's Law predicts it's pretty hard.
In principle, I agree--right down to the likelihood of Murphy doing his utmost to interfer.

My Mitutoyo 100x/0.70, which I quite like and use frequently (it has 6mm WD), has, on the back, an opaque black plastic with a center hole of about 3mm diameter. I assume this is a flare-cut stop, and the image-forming light travels through that 3mm aperture.

I have no idea if the Mitutoyo HR 100x/0.90 has a similar rear stop, but if it does, I'd likely find a 3mm aperture difficult to create a lighting gradient through. (As Rik predicted.) If this were a lens I already owned, I'd try all kinds of things to make it work. Or if this lens were inexpensive, I might have a go and try to make it work. But $950 USD is more than I'm willing to risk on an experiment that seems likely to disappoint.

Macro_Cosmos, your preference for cube beamsplitters is notable. Of course cube and plate beamsplitters each have particular virtues and vices. But a couple of involved forum threads posted a few years ago convinced me that plate beamsplitters offered more attractive trade-offs for my applications. Further, a derelict, formerly high-end, microscope assembly that I bought cheaply for parts contains a couple of nice plate beamsplitters. And my Mitutoyo FS-70 microscope head (not yet integrated) seems to contain a plate beamsplitter. So my intent is to prefer plate beamsplitters unless conflicting experience changes my mind.

Cheers,

--Chris S.

Macro_Cosmos
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Post by Macro_Cosmos »

Chris S. wrote: Macro_Cosmos, your preference for cube beamsplitters is notable. Of course cube and plate beamsplitters each have particular virtues and vices. But a couple of involved forum threads posted a few years ago convinced me that plate beamsplitters offered more attractive trade-offs for my applications. Further, a derelict, formerly high-end, microscope assembly that I bought cheaply for parts contains a couple of nice plate beamsplitters. And my Mitutoyo FS-70 microscope head (not yet integrated) seems to contain a plate beamsplitter. So my intent is to prefer plate beamsplitters unless conflicting experience changes my mind.

Cheers,

--Chris S.
I've had all kinds of alignment trouble with the plate beamsplitters, however now I have both, I'd soon find out which is suitable for me.

A repurposed microscope head with everything pre-configured and the light path tuned would be easier to work with. I was recommended that but I would like to try out modular cage systems. I can also add components without much thought. I want to build a Köhler illumination setup too, those modular cage systems look really fun to experiment with.

I purchased a pellicle beamsplitter for cheap, maybe that would work out well?
https://www.edmundoptics.com/p/254mm-cl ... tter/4504/
Fragile, they are very thin, maybe they are better than both cube and plate BSs?
Absolutely no idea how they work. 40:40, where does the 20% go?

It's just like the cube BS, but without the 2 glass prisms. Could be the ideal choice!
Image

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Post by Pau »

There are some approaches to avoid on axis epiillumination like the Zeiss h-Pr illuminator which I only know from reading
at Cebulla, W. Handbook of Incident Light Microscopy (Zeiss), pg. 11-12

Image
Pau

Horst_S
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Re: Advice please: High NA air objective with small WD

Post by Horst_S »

Chris S. wrote:
Mon Apr 06, 2020 5:06 am
...
Can anyone with knowledge of axial illumination (if I'm using this term correctly) help me gain a sense of how much additional information, if any, this lens, plus system modifications needed to use it, would offer? ....
Hi Chris,

is this still from interest for you?

Illumination through the lens is typically used in metallurgical investigation/material science/quality inspections.. (I think the "M" often found on objectives for reflected light derives from "metallurgical").

With the appropriate technic (brightfield/darkfield;polarization,DIC) it can -for example- detect defects in surfaces that are otherwise hard to see. It is also possible to look into holes.

Cross polarization can eliminate even extreme reflections -one fixed polarizer is mounted in the illumination path before entering the beamsplitter, a second one (turnable) between beamsplitter and tubelens. So the extinction can be adjusted in a wide range. It is possible the get high contrast on many objects. On low magnification "M" objectives you can sometimes find an additional polarizing, turnable front element.

Is there any object particulary from interest for you?

PeteM
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Re: Advice please: High NA air objective with small WD

Post by PeteM »

Many reflected microscopes offer a darkfield option. It won't help, of course, with an objective not intended for darkfield (with an illumination cone surrounding the objective). But it might be a way of getting images from an alternative high NA dry objective -- with less of that flat "deer in the headlights" look.

It also might be interesting to see if edge-lit led lamps could provide an alternative to trying to find space for fiber optic guides in sub-1mm working distances. Some of the very cheap (under $20) edge lit LED tracing tablets are around 1mm thick. Could be they would survive a hole put through and a bit of machining to make an objective surround with light coming in from some fraction of 1mm thickness edge lit??? Wouldn't be real bright, but a bit of side lighting might be useful?

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Re: Advice please: High NA air objective with small WD

Post by Chris S. »

Horst,

Yes, this is still very much of interest to me. One thing I’d like to do is look into holes as you mentioned. For example, in my post on Papilio ulysses wing scales, I’m fairly sure the black spaces in the lattice work are not through-holes, but light-absorbing dimples or shallow pits. Even though I made nearly the entire ring of space between objective and subject a diffuse light source, this light was too oblique to shine into these dimples. The only way I see to shine light into the dimples is to project it through the lens.

I find this a fairly common issue with high-magnification Mitutoyo lenses—their wide, blunt ends may leave sufficient room for light to shine around them, but this light necessarily comes from such a glancing angle that it doesn’t shine into the deeper recesses of some subjects.
To avoid the flat, “deer in the headlights” look others have warned of, I’ll likely, in most cases, mix light from outside the objective with light sent through the objective; in traditional photography terms, the outside light will be the key light, and the through-the lens light will be fill light.
Cross polarization can eliminate even extreme reflections -one fixed polarizer is mounted in the illumination path before entering the beamsplitter, a second one (turnable) between beamsplitter and tubelens. So the extinction can be adjusted in a wide range. It is possible the get high contrast on many objects.
Thanks for this. I’d been thinking about trying cross-pol here, appreciate your description, and welcome any further thoughts you have.

Toward adding through-lens lighting to my macro rig, last week I ordered an Edmund Optics Techspec C-mounted plate beamsplitter. I plan to mount it removeably on my tube lens assembly:

Image

The housing for the Edmund beamsplitter has C-mount-threaded openings, and my tube lens assembly is made with Edmund parts that have C-mounts, so the beamsplitter should be fairly easy to integrate. I have some neutral, high-extinction polarizing sheets from Edmund Optics that should work for the light source. And I have a C-mount to Mitutoyo-mount turnable Edmund Optics polarizer (also with good neutrality and high extinction) to go between beamsplitter and lens. (I’ve been using this combination for years in cross-pol with standard reflected lighting.)

Question: Do I need to collimate the light source? I was planning to experiment to find out, but you likely know the answer already.

You mentioned DIC. What I’m looking to assemble, here, is a good portion of the way to epi-DIC—“all” that’s wanting is the prisms and something to hold them properly. (Those used to transmitted DIC might be surprised how much simpler an epi-DIC setup is, at least according to diagrams I’ve seen.) I have several Nomarski prisms intended for an old Nikon system, and have thought I might experiment at using them with my Mitutoyo objectives. But all I know about epi-DIC is what I’ve read—I have no hands on experience with it, nor even a clue how the images should look like if I get it right.

Thoughts you have on any of this are indeed welcome.

Pete, Pau, and Macro_Cosmos, thanks also for your input. I’ve addressed above why I want something in addition to edge light. It’s not hard to shine sharply oblique light in around the objective front, but the modeling and ability to shine into pits and crevices is sometimes wanting.

Macro_Cosmos, I have to say that pellicle beamsplitters have struck me as very unhandy for our work--expensive, thin, fragile, and hygroscopic. Recipe for disaster, outside the confines of a sealed instrument? Also, I don’t see much harm in the offset of rays created by a plate beamsplitter in what I do. Am I missing something?

Pau, those diagrams from Zeiss are both fascinating and daunting. I may return to them if the simple beamsplitter with cross-pol, plus external lighting, does not suffice.

Cheers,

--Chris S.

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Re: Advice please: High NA air objective with small WD

Post by Pau »

Question: Do I need to collimate the light source? I was planning to experiment to find out, but you likely know the answer already.
As usual in Microscopy the ideal illumination is after Kölher: focusing the light source at the objective rear focal plane filling it. This is what epiilluminators do, both for epiilumination and epifluorescence. You could also play with it up to some extent it to get oblique illumination.
Just collimated it will provide more or less critical illumination, I think.

Reflected DIC, AFAIK, provides color contrast of metallic reflective subjects, again very different than transmitted DIC, although I have very little experience, see viewtopic.php?p=154251#p154251 and the links inside
Pau

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Re: Advice please: High NA air objective with small WD

Post by Scarodactyl »

The Nikon epi DIC prisms I have tried (from the finite Optiphot series) give good results with mitutoyo objectives, at least when set up on a Nikon microscope. I'll eventually try it with my newer eclipse-era DIC slider but I need to have an adapter made to mount the nosepiece. Subjects don't have to be metallic, just reflective.

That said depending on the geometry of the subject you might see a bit of frustrating ghosting/image doubling. This is present with the original Nikon objectives as well, and seems to just be inherent to how dic works.

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