Nikon's 60x oil CF N Plan Apochromatic.

NikonUser · Post by **NikonUser** » Thu Nov 25, 2010 1:59 pm

Finally upgraded my Photoshop CS2 to CS5.
Used this 60x on an Olympus microscope with an Olympus 2.5x NFK relay eyepiece.
Smallest Desmid I could find was actually too large for the 60x and 2.5x relay lens on a (23.6 mm sensor); but I wanted to try and see what this lens could do with a less than optimum setup (but it's the only one I have).
7 RAW (Nikon NEF) frames @1µ, corrected for white balance and chromatic aberration in CS5, converted to jpg's and stacked with ZS PMax.
I suspect the clear halo around the desmid is a layer of mucilage.
This lens is obviously usable for reduced-sized images.
Top: full frame
Middle: actual pixel selection
Bottom: cloned image

NUM10107

Mitch640 · Post by **Mitch640** » Thu Nov 25, 2010 2:27 pm

You have explained to me before, how you clean up your images. Did you clean up each image before the stack? I suspect you would have to, although I have not used the software yet. And how do you like CS5?

Your final image of course, is better than anything I have gotten yet, but I think I have a couple weeks to go.

NikonUser · Post by **NikonUser** » Thu Nov 25, 2010 2:55 pm

The cleaning up was done in my plate with 12 depressions; just kept moving the desmid from depression to depression, each time leaving behind all the other stuff. Just changed the WB and corrected some of the CA before stacking.

First time to use CS5, and am at the bottom of a steep learning curve.
Fortunately the upgrade left my CS2 intact so I further processed the ZS stacked image in CS2; could not find a way to see more than one image on the CS5 desktop; CS2 lets me have as many images open all at once.

(Save your money Mitch, when I can find a 60x Olympus S Plan Apochromat do I have a great deal for you on a 60x Nikon)

rjlittlefield · Post by **rjlittlefield** » Thu Nov 25, 2010 3:04 pm

NikonUser wrote:could not find a way to see more than one image on the CS5 desktop

Have you tried messing around with Window > Arrange > Float All in Windows?

--Rik

Mitch640 · Post by **Mitch640** » Thu Nov 25, 2010 3:06 pm

I just upgradded to CS4 and I suspect that CS5 is similar, both a lot different than CS3. When CS4 open multiple images, it's in one frame with a bunch of tabs at the top. Click on the one you want to see by itself, then grab it by the top bar and peel it off the main window and let go, now you have a separate window with that one image in it. It helps to have dual or more monitors. I use a dual widescreen setup that makes this easy to work with.

If you didn't get that working, I was going to offer, but it will take some time to save that much. Although, the more work I do on my fill in job, the faster it will go. Now I have a reason to grind the nose. LOL

NikonUser · Post by **NikonUser** » Thu Nov 25, 2010 3:16 pm

Rik: Haven't tried to do much with CS5, had it for only 24 hours.
Will try what you suggest.
I hate online manuals so bought a CS5 "How To" book, it's only 1.042 pages

Mitch: CS5 seems to be quite a bit different from CS2. I was surprised that Adobe would let me upgrade from CS2 for $250.00 Canadian (something like $800.00 for the full verison for 1st time buyers.)

Mitch640 · Post by **Mitch640** » Thu Nov 25, 2010 3:32 pm

Yeah, the upgrades are more than reasonable. Buying that first one is expensive, but they make it easy after that. And I have found, well worth the cost. I use it so much that I have to keep it upgraded.

René · Post by **René** » Fri Nov 26, 2010 1:36 am

Why don't you try it without the projective? Looking at the middle picture, you won't need it. And that's also my impression from the new infinity line, where I got our simple cmos cam directly coupled to the photoport without projectives (besides of the tubelens, but this is not necessary for the 160mm CF one). What's the NA of this lens, 1.4?

Best wishes, Rene

Mitch640 · Post by **Mitch640** » Fri Nov 26, 2010 4:32 am

Why don't you try it without the projective?

I can attest to this working. It will actually focus without a projector lens in there, but it won't be parfocal anymore. Not a problem with live view anyway.

NikonUser · Post by **NikonUser** » Fri Nov 26, 2010 7:02 am

Rene: I don't believe it would work.
As Pau mentioned
IN THIS POST
NA 1.4 objectives are incredibly sensitive to tube length.
This objective 'needs' some sort of image at 150 mm from its 'base'.
This position is down inside the phototube of the BH2.
The bottom lens of the NFK projection lens intercepts this image and projects it 125 mm above the top of the trinoc. tube where it can be intercepted by a camera's sensor.
Placing the camera sensor at the top of the trinoc phototube (no projection lens) will mean that the tube length is now about 20 mm greater than optimum.
You can see what effect that has on image quality in the above reference.

I suspect your method would be OK for lower NA objectives, these are more forgiving regarding tube length.

Mitch640 · Post by **Mitch640** » Fri Nov 26, 2010 8:02 am

Can't you adjust that with your bellows setup?

NikonUser · Post by **NikonUser** » Fri Nov 26, 2010 8:21 am

Not sure what you mean, Mitch.
The 'problem' with high NA objectives (1.4 in this case) and the BH2 scope is that the image created by the objective is at its highest quality somewhere down INSIDE the trinocular tube. It is physically impossible to get your camera's sensor inside this tube.
Sure, you can get an image at various distances above the tube but the further away from this magical point inside the tube, the crappier the images get. One doesn't spend this type of money to get crappy images

Mitch640 · Post by **Mitch640** » Fri Nov 26, 2010 8:54 am

Ah, I see just what you mean now. I guess I was so use to crappy images, I could not see the difference between bad crap and fair crap. Now I just played a little more with my cheap extension tube adapters, and the obvious is becoming more clear. While I can now, I assume anyway, get the bottom of my Projector lens at the correct distance, by making sure there is nothing in the way of it bottoming fully into the tube, I just found out that the distance from the top lens of the projector lens to the sensor is also important in determining the quality of the crap. How would you go about measuring that? I know about the mark on the camera body for the sensor plane, but how can you reliably measure it to the surface of the Projector lens. If you do measure to the surface of that lens and not down inside it? And what is that distance supposed to be?

Pau · Post by **Pau** » Fri Nov 26, 2010 11:11 am

Mitch640 wrote:Can't you adjust that with your bellows setup?

It can be done for sure, but removing the trinocular head and placing the bellows directly over the microscope frame without photoeyepiece. It will work OK, in fact it's the same that using it on bellows, a very popular setup in the macro forum with low magnification CF objectives, but with a 60X 1.4, altough doable, the lost of the direct view don't seem very practical.

NikonUser · Post by **NikonUser** » Fri Nov 26, 2010 11:36 am

Pau is correct, I never thought of removing the entire trinoc head.

Olympus specify a distance of 125 mm from the top of the yellow phototube to the camera sensor. That is, put a NFK relay lens in the tube and then measure from where this lens rests on the phototube of the trinoc. So that's easy to measure.

Have yet to find what Nikon recommend for the CF PL relay lens.