Leica S8APO to Canon 1000D

[ChrisRaper]

I've been fortunate enough to get hold of a Leica S8APO recently - a lovely microscope with exceptional resolution. The suppliers also provided a fine-focus stand and kit of bits to connect-up my Canon 1000D but the quality of the image stacks I am getting is much too soft for my liking at high magnifications and I was wondering if anyone has mastered this before and might have some suggestions.

Here is the camera and tubes:


... and here is an 80x stack of a fly spiracle (using flash) - the hairs just come out fuzzy no matter how I do it:


I'm not too sure whether I am expecting too much but the image that I see in the microscope eyepieces is much clearer. Also, the camera kit contains its own optics - one of which houses a WF 10x microscope eyepiece - so I am not to sure whether this eyepiece might be letting the system down.

Any advice would be great

[seta666]

I used a 100D for stacking in the past, you have to bear in mind that the 1000D mirror moves between actuation so enought time for the sytem to settle must be allowed.

About the optics I can not help

Regards
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[Cactusdave]

I'm no expert, but there looks to me as though there may be several problems going on. The microscope is an excellent one and you are most fortunate to own it. I have been looking for a stereo of that quality for some time, but prices were too high. That 'scope is capable of delivering much better images to your camera.

I think the previous poster is right that you have a vibration problem, the shutter being one factor. Also is the stand supplied completely stable and capable of giving the very small focus increments needed at this magnification reproducibly and without slippage?. Finally I'm not sure you have optimal lighting. You say you are using flash. Is that a single flash or a pair? How are you diffusing the light? In my limited experience stereos are better with continuous illumination. Ideal for incident illumination is a fibre optic pair suitably diffused.
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Leitz Ortholux 1, Zeiss standard, Nikon Diaphot inverted, Canon photographic gear

[ChrisRaper]

Interesting, mirror movement wasn't something I had factored into the equation - I might try using Liveview to lock it up. Wouldn't the fact that I was using flash have frozen the image and made the mirror irrelevant though?

@Cactusdave: The rack stand seems to be pretty stable - I haven't seen any drift over time and it is quite stiff to focus. The fine-focus adjuster moves 0.56mm in a full rotation so it should be fine enough to give good steps at this magnification. I have taken single-shots and they are fairly fuzzy too so stepping distance isn't an issue.

The flash is just a single unit and wasn't diffused on those photos but it had been bounced off some foam surfaces to flatten it out a bit.

Feeling very privilaged to have one too I have never had the money to afford equipment like this but I was fortunate enough to qualify for a government grant that supports invertebrate recording in the voluntary sector. More on the story here.

[seta666]

The 1000D mirror and shutter motor are the same, so everytime you shot mirror moves, even in liveview.
the XXD series have different motors for mirror and shutter.

Also, if you use flash use it in second curtain, mirror lock up and 2" or more exposure time, at high magnification (40X and such) I would use 3-4 " settle time.

Also the EFCS does not work when using flash, again allow enough time for shutter vibrations to settle

With continious light you can use EFCS, but between exposures allow those 2-4" for the mirror vibrations, once everything is still you can use sort exposure time

Here you can see some images taken with the 1000D, obviously a different set up but can serve you as a guide of what to expect of the 1000D sensor

http://www.flickr.com/photos/seta666/tags/eos1000d/

Regards
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www.macrosmuymacros.com

[ChrisRaper]

@seta666: Wow ... stunning images!

I've had fairly good results so far using directly connected finite objectives, on bellows and with things like reversed enlarger lenses ... it's only this S8APO that seems to be the problem but microscopes are a completely alien world for me so I am sure that there is plenty more to learn.

I will switch the 1000D into second curtain flash and 2" exposure time ... but I'll wait until I can black-out the room, tonight Finger crossed!

[rjlittlefield]

Chris, is the image that you've posted full frame, actual pixels, or something else?

When comparing eyepiece and camera views, be sure you're comparing apples and apples. It is easy to get misled by comparing the overall view through the eyepieces against a tiny crop that you see at actual pixels in the digital image. Check carefully to see if you are actually able to resolve more detail in the eyepieces than you can see in the digital image.

I would expect flash to kill most vibration problems, even with first curtain sync.

--Rik

[seta666]

Also a 10X eyepiece for photography seems like too much for me, seems like adding a 10X teleconverter to any lens
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[Pau]

[ChrisRaper]

@Rik: the image I posted was the best end result that I could obtain - after lots of PhotoShop sharpening - sorry for the confusion. I will fish out some single-shot full-frame crops.

I take your point about comparing apples ... my understanding is that the objective produces a 1x image and then this passes through the zoom which multiplies it up by 8x and then the eyepieces multiply that by 10x to get 80x. The camera tubes pick off the image after zoom (so 8x) and this passes through a few 1x tubes and then into a 10x eyepiece and the sensor seems to be resolving an image that is very similar in coverage to the eyepieces.

I am sure that in the eyepieces are resolving more detail - in the case of the spiracle I can clearly see the hairs but in all of my images they seem slightly fuzzy in comparison ... sorry, not very objective, I know. I also checked it against other hairy surfaces and I just never see the fine detail as crisp as I see them in the eyepieces.

@seta666: I have a 5x eyepiece from the same no-brand manufacturer so I can try substituting that but when I tried it last the image on the sensor was very vignetted.

[ChrisRaper]

[ChrisRaper]

Just for reference, here is a quick n dirty full-scale crop of a stack of a butterfly wing - taken with the scope set on 80x. No sharpening or other post-processing.

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[rjlittlefield]

At "80X", what is the field width seen by your camera?

--Rik

[ChrisRaper]

[seta666]