MicrobeHunter.com Microscopy Forum

I'm only Nikon, so here's a list of Nikon scopes that match your criteria.

1.S-Ke (L-Ke)
2.Apophot
3.Biophot
4.Microphot (SA,FX,FXA)
5.E600,E800,E1000

Not aware of staining that would work for whole mounts... maybe silver staining could work since it's used for fairly thick neural tissue. If it could be sectioned, H&E would be the preference. If you have access to a fluorescent microscope, or a confocal/light sheet/MP microscope, fluorescent stain...

I've never done plant/fungi specimens, but used Safranin-O and fast green FCF for staining cartilage sections fixed in PFA. If it's about the Safranin-O staining intensity, changing the solution pH (I used sodium citrate buffer) would improve things. Higher pH would yield more intense staining, but ...

Yes, Pete's comments already nails most of what is needed to be said. No, that kit won't fit the Microphot SA. The nosepiece holding the upper prism and analyzer will only fit the Optiphot-2 and some very late original Optiphots. However, the prisms and condenser are common components for the Microp...

Umm, that DIC system photo actually belongs to me. I've granted use for a person who I won't name here, but I haven't given up copyrights for it.

Cheers,
John

Most phase turret condensers have mediocre NAs compared to other high-end condensers. Firstly due to lack of need to illuminate the absolute periphery of the available NA (most phase plates are only at the middle range of NA), and secondly to have longer working distances (especially the one you men...

Not sure with LOMO, but some manufacturers have different parfocal distances for low power objectives, especially for those under 4x.

Cheers,
John

Perhaps cost?
Another would probably be field curvature that cant be compensated with the lens itself.

Cheers,
John

How large is the sensor? If sensor size is available, just divide it by the number of pixels on either axis and you'll get the approximate actual size of each pixel. Next imaging a stage micrometer will get you the total system magnification. A microscope camera should have its pixel size in its spe...

Erm, wouldn't Thorlabs confocal system be a laser scanning confocal?? That's different from a spinning disk confocal.

Ah! Do you have a DM IL??
That slider slot under the head accepting the Hoffman sliders is actually conjugate with the objective BFP...

Cheers,
John

Well... in a infinite system with a spinning disk confocal setup you should have the pinhole disk "after" the tube lens (or secondary objective, in Nikon jargon) That pinhole at the primary image plane eliminates light from the other defocused planes and at the same time acts as the illuminating ape...

BTW, Nikon does it like this in their "External Phase Contrast" system on their inverted microscopes.

The BFP is the Fourier plane of the objective :-) So if the BFP is at the left of L1, it will be relayed to the right of L2. The 4f relay is normally used to Fourier transform the "image" made by the objective by L1, and do a following inverse Fourier transform with L2. If you're relaying the BFP wi...

Yes, that 1987 patent would be a valid description of how the U-CPA works. And that 4f relay in the Bertrand position will kind of work... At least the Fourier plane wouldn't be at the center of L1 and L2. If the focal plane f1 to the left of L1 conincides with the objective BFP, the BFP will be rei...

>Pete
All manufacturers should publish that BFP location data! Olympus does and as you mention Leitz does... but Nikon doesn't disclose that data (at least on the www.)

Yup, the paper from the Zia lab (Fig.1c) illustrates how a 4f-relay is implemented. Fig.1a has a Bertrand lens behind the tube lens, in this case consider the objective+tube lens as a unit functioning as a finite objective. Looking at it in that light will fit in with the 1966 patent by Leitz you sh...

One needs to know where the conjugate planes reside. In your case assuming Koehler illumination is set up, conjugate planes would be like this; light source>condenser iris (condenser side prism interference plane)> objective exit pupil (objective back focal plane, objective side prism interference p...

Excellent stuff! I just got a 2oz tube of 206, and
it works wonders. I've went as far as relubing most of my objectives with correction collars, no sticking again! (hopefully)

BR,
John

Are the prisms matched to the objective?
Shear amount should be tuned to be a tad under the objective resolution limit.

BR,
John

Excellent reference, the original document by Nomarski is rather cryptic (my French is horrible.) Kusaka describes the essence of prism design in the patent. One thing worth noting is Fig.7A-8. The shear amount is physically dictated by the wedge angle alpha (as in Fig.3), and the focal length of th...

With these kind of hemispheric top lenses, you have to consider "how thick" your slide glass will be. Note the tight tolerances for oil DF condensers.

BR,
John

Viktor, thank you for the Paolo Pozzi reference. I had some difficulty in determining objective BFPs on my bench so this method is excellent.

BR,
John

Some corrections and follow ups. Finite CF metallurgical objectives (M Plans) by Nikon are tube length(TL) 210mm. TL160mm pre-CF metallurgical objectives exist, but they are the short barreled variety and will have parfocality problems. Regarding the mounting thread, M Plans are RMS so they will mou...

The field iris should be set to the smallest FN in the system. If you use a FN22 ocular on a FN26.5 head, it would be FN22. Many heads (and objectives) will allow fields up to nearly 30mm only limited by the ocular tube diameter themselves but whether that periphery would be pretty is a different ma...

You’ll need a vertical facing filament to fill the aperture at high numerical apertures.

Cheers,
John

That objective has a safety spring nose, so it's essentially a lens holder inside a outer tube pressed against a forward flange with a spring. You have to access the inner lens assembly from the rear (mount side) loosening the ring screws. Though you'll eventually get inside to the elements you susp...

That reverse revolver adapter is a late addition to the original Optiphot. Though the revolver is reversed, there's no supplemental lenses (neither in the revolver, nor the stand limb) in the Optiphot2,and Labophot2 due to a redesigned thinner limb compared to the original Optiphot. Thus no addition...

Try mica. It used to be the material of choice

Cheers,
John

Darkfield is inherently lower resolution than brightfield, and also is not a faithful representation of the specimen image, due to the loss of the zero-order light in the fourier-reconstructed (via interference) final image. Carl Partly agree, partly not. Please correct me if I'm not understanding ...

"Well, why not get a 10x rather than a puny 2.5x when you can!"

I think that's the rationale behind it, and many would eventually come down to lower mags when they realize what resolution is

Cheers,
John