Images made through a microscope. All subject types.
This is my first real attempt at stacking. Subject is a 2mm long (actual) spider (unknown to me). Method is: S series Nikon microscope, PB-6 bellows, Nikon Po 4 0.1 objective, Nikon D300s, direct lighting from dichroic (qi) light-source + small reflector. This image produced from a stack of 89 images guessed at around 30 micron intervals with minimal retouching. I am still working on better consistency of intervals at the moment but think the problem I will have is, the S series microscope's fine focus does not work. Keep me in mind if you see another S series around-say on EBay for instance. I think this also could have been produced better by reduced increment spacing say 20microns and diffused lighting.
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This is definitely a good first effort. I agree that more diffuse lighting would help. The microscopyu DOF calculator says that DOF for a 4X NA 0.1 objective is at least 65 microns, so 30 microns focus step seems fine. I'm bothered that the center looks clean but the edges look soft and the lower left corner looks radially smeared with color fringes. I'm wondering if that particular objective is one of the older Nikons that was not CF.
You are probably right. This objective came with the microscope sold as a decomissioned instrument so the objectives may never have been up-graded since new. The objective is marked Nikon 12734 LENS MADE IN JAPAN. I have a little Vickers England 5/0.15 (looks well used-usually a good sign) objective I want to try next, this time with more precise focus intervals trialing a system I have come up with. Watch this space.I'm bothered that the center looks clean but the edges look soft and the lower left corner looks radially smeared with color fringes. I'm wondering if that particular objective is one of the older Nikons that was not CF.