Spectacular as always. Its like you have an optical version of a SEM. Its beautiful shots like this that make me want to try stacking. Of course its the full cabinet of histological samples waiting to be sectioned etc that stop me...but I am still curious about the details of your rig and conditions. How many images / step size etc. did you use?
David
Fly ("details")
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Charles Krebs
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Rik:
With the 50/0.50 the DOF is still ridiculously small, so typically much more time is taken carefully adjusting the subject into the the position that makes best use of what DOF exists.
David:
The set-up is the same I've used for awhile now (picture below). While I sometimes use the Ikea lamps for lower mags, the exposure times needed for the 50X gets too long for my taste. I typically use fiber-optic illuminators with (several light guides) with the 50X.
I've used flash before, but I have found that it is easier to "finesse" the lighting when using the "diffusion dome" with several small continuous sources.
Marek:did you have to use the trick of shooting extra images with a stopped-down aperture to retain enough visible structure in the OOF areas?
The first image was taken with an Infinity Photo-Optical TS-160. And the other two with the Olympus 50/0.50. In both cases the maximum apertures are a little on the small side if you are going to pixel-peep at 100% (and compare to higher aperture optics). But for screen size images (up to about 1500 pixels wide... give or take) with judicious sharpening, the final result is hard to distinguish from the much larger aperture optics. And since the DOF is larger than a high NA objective, it makes stacking much easier, especially when you have the crossing hairs and bristles at different depths. The other benefit is that (as you observed Rik) it keeps the out-of-focus areas a bit more "together" with less of that drastic DOF drop-off.What objective have you used in the first photo ?
With the 50/0.50 the DOF is still ridiculously small, so typically much more time is taken carefully adjusting the subject into the the position that makes best use of what DOF exists.
David:
31 images in the eye stack. This is a very low number for a 50X stack, but really there is just a small band that is sharp. 91 images in the "bristle" stack. In both cases the step size was just a little over 2 microns.but I am still curious about the details of your rig and conditions. How many images / step size etc. did you use?
The set-up is the same I've used for awhile now (picture below). While I sometimes use the Ikea lamps for lower mags, the exposure times needed for the 50X gets too long for my taste. I typically use fiber-optic illuminators with (several light guides) with the 50X.
I've used flash before, but I have found that it is easier to "finesse" the lighting when using the "diffusion dome" with several small continuous sources.
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RogelioMoreno
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