Wing scales, 3 stereo pairs @50X

[Charles Krebs]

For these I went from using an Ikea light to one of my fiber-optic illuminators since I really wanted more light with the 50X objective. But I am always a little worried about having a fan-cooled illuminator on the same table as the microscope (because of some vibration "issues" on a couple of occasions in the past). So these were made partially as a test. For 2-dimensional images these are not that spectacular but I like the way they come to life in 3-D.

The top is from a Purple Spotted Swallowtail (Graphium weiskei). The other two are from a day flying moth, Green-banded Urania (Urania leilus).

All were taken with the same set-up:
Nikon MM-11 microscope, Olympus 50/0.50 LMPlanFL N, Canon T3i. 50X on sensor.

The field area of each image is 0.3 x 0.45mm (0.012 x 0.018 inch).

[pierre]

Great Work Charles

Such small field is something very hard to deal with (it still a dream for me).

Congrats.

I am a bit curious.
Is the shutter of your T3i completely silent like the 50D and do you proceed using raw format of pictures ?

[rjlittlefield]

Some curl up; some curl down; some have ridges like Ruffles.

All very much more "alive" with the stereo. Lovely!

--Rik

[Chris S.]

But I am always a little worried about having a fan-cooled illuminator on the same table as the microscope (because of some vibration "issues" on a couple of occasions in the past).

Charlie, I've had some issues with this too. Now my illuminators hang from ropes suspended from the ceiling. This has turned out to effectively prevent vibration produced by the illuminator fans from effecting images. And as a side benefit, it places the illuminators in convenient positions for my shooting. On the other hand, it looks as if I forgot to pay my gravity bill.

Capital images, Charlie.

--Chris

[arturoag75]

As usual, excellent

[Wim van Egmond]

They are brilliant. The second one has a nice transparency.

I wonder if it would help if you would put the light source on a pillow, or thick foam. I will try it.

I am thinking of buying a Canon. I have use Nikon but always have to use flash or very long exposures to avoid the shutter movement. Any advice which type to choose?

About the curling of the scales. Could it be that they curl when they dry?

Wim

[pwnell]

These are beautiful! How did you manage to not get stacking artifacts? If I stack at high magnification I end up with 200+ images, and always struggle with out of focus areas not rendering correctly. Is it because of the low NA of your 50 vs. my 0.95 on the 40x UPlanSApo? I guess that objective is not right for the job...

Your technique always astounds me. Great work.

[canonian]

Incredible shots Charles, 50X is way beyond my reach or capabilities.

... Any advice which type to choose?

Hi Wim,
As a long time Canon user (hence Canonian ) I would say a 650D on the budget side or a 5DMkIII on the more expensive side.
Depends weither you're looking for APS-C (DX) or Full Frame.

Not all the lenses you are using now will play nice with FF.
Can't remember which Nikon you shoot with now.

To make the choice harder there's also the lovely Sony NEX'es.

[Charles Krebs]

Thanks all for the comments!

pierre,

I am a bit curious.
Is the shutter of your T3i completely silent like the 50D and do you proceed using raw format of pictures ?

No, the T3i makes a faint "squish" sound at the start of the exposure, but I have not seen any vibration occurring at that time. Most of the Canon bodies make a similar sound from live-view, the 50D is the only totally silent one I have seen. The 60D does show some vibration at the time of the slight sound. While I have not actually tested one on a microscope, I suspect the full frame 6D might also have a very slight vibration as well. (This is based solely on an in store, hand held "evaluation" and so I hesitate to even mention it. But if I were looking for a full-frame body and completely vibration free EFSC were a critical feature it needs to be checked out more carefully).

I'll confess that very frequently the out of camera jpg's are used. But in this case all three were done from raw converted to TIF files. A 50/0.50, even an excellent one, is at about the limit (past it actually!) to obtaining a reasonably sharp looking image (effective aperture being about f/50!). So the little bit of extra "sharpness" that can be squeezed out of a raw seems to make a difference. Especially on the fine lines on each scale most noticeable on the first two images. (In reality, since resolution has taken a big hit from diffraction, the slight improvement likely comes from avoiding the JPEG stage and being able to choose the best balance between "sharpening" and noise reduction).


Wim,
I am only completely confident talking about ones I have actually used. The 50D and T3i (600D) are OK in this regard. (While I have not tried them I think it is likely that the T4i/650D and T5i/700D are the same as the 600D. But they should be checked out before purchase).

Based on reports from people I trust, the 7D and 5DII (full-frame) and 5DIII (also full-frame) are OK as well.

The 60D has some slight vibration. A 70D should be introduced this summer, but it obviously remains to be seen if it will have any "issues" in this regard. As I mentioned above, I am suspicious of the full-frame 6D, but have not really tested one.

One nice feature of the mid and upper models (those above the Txx/xxxD) is that they use separate motors for the mirror mechanism and the shutter. This means that you can operate from live-view with the mirror up at all times, with only the shutter "re-charging" between exposures (in essence, much like a mirror-less camera). The more basic models (and most SLRs) have a single motor, so after a shot both the mirror and shutter must cycle together. It doesn't cause more vibrations in the picture (unless perhaps you run off a rapid sequence of shots with no "settle time" at all), but it does make for a quieter, "gentler" overall operation when shooting an image stack.

I've often wondered about the curl in the scales myself. Without the opportunity to see a live specimen I don't know. I could put the sample used in a "relaxing chamber" and see it there is any change, but even that would not be a definitive answer. Perhaps some forum member with more knowledge could offer some insight. I would be curious about how "hydrated" scales are on a live specimen.

Chris S.
I've contemplated suspending the FO illuminators from the ceiling and would probably go that route if vibration were a problem. (This would free up more table-top "real estate" as well). My workspace already look like a sloppy mad scientists den, so that would only add to the ambiance.

Waldo,
There is no doubt that it will be easier to get fewer stacking artifacts with a 50/0.50 than with a 40/0.95, although you do give up something in terms of resolution. I'm dealing with a dry, uncovered specimen so I don't have the subject compression often afforded by a cover slip. Plus I need the working distance above the subject to allow for the type of lighting that is preferred. So the trade-off of the smaller aperture is a little more palatable (at least that is what I tell myself ). Also, if you look at the sections photographed you'll see that they were selected to have minimum open space between the uppermost part and lowest part. Even though these were shot at 50X (with an admittedly small aperture) the number of images required were 44, 43 and 34 respectively. Quite a modest number. (All it would take is one scale jutting upward to double or triple the number of imaged needed and also likely cause artifacts in the result).

[Ozelot]

Hello Charles,

very nice pictures.
Here, the structure of the individual scales is very beautiful to see.
Top quality.

Greetings

Michael

[Chris S.]

I've contemplated suspending the FO illuminators from the ceiling and would probably go that route if vibration were a problem. (This would free up more table-top "real estate" as well). My workspace already look like a sloppy mad scientists den, so that would only add to the ambiance.

Charlie, I understand your viewpoint very well. I'm not certain that my FO illuminators really need to be suspended from the ceiling--it was something I tried early in my experiments. But since it's worked nicely, I haven't checked back to see if it truly was necessary. Maybe I, also, rather like the "Mad Scientist" vibe. But also, the illuminators, when hanging, are in positions that make them very convenient to use--and each FO illuminator, hanging from its rope, is free to rotate such that the FO light guide can point in exactly the direction needed--very handy! And the table-top real estate saved--as you mentioned--is much appreciated.

Also, I have solenoid-activated shutters on some of my FO illuminators, and these solenoids can make a thunk when opening or closing the shutter So I don't mind the extra mechanical isolation that prevents these thunks from being conducted to my subject or camera. (Of course, it would certainly be possible to rig more gentle shutters for continuous FO illlumination--I just haven't done it.)

--Chris

Edited typos

[Wim van Egmond]

Thank you Charles for the extensive comments! And thanks Fred. It is a pity that Nikon can't shoot from live view the way Canon does. It would be nice to have a very fast burst mode from live view. But I guess ghat is not possible yet?

I'll try to post some images this week. I have new material but I have changed to a new computer so I am trying to improve the work flow.

Wim

[Jacek]

I do not see why the stereo effect?

[RogelioMoreno]

Charlie,

Excellent!

Rogelio

[Charles Krebs]

Jacek,

These are cross-eyed stereo pairs. With a little practice most (but not all!) can "free view" them to see a stereo view.