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A Single and Double Diatom
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Mitch640



Joined: 15 Aug 2010
Posts: 2137

PostPosted: Tue Aug 31, 2010 12:24 pm    Post subject: A Single and Double Diatom Reply with quote

I've been practicing, getting things working on the new scope. I found that I apparently have the wrong adapter for the third tube in the trinocular scope, so I pulled an eyepiece and put the video camera in that, and started getting better images.

All I know is, these are diatoms. Not sure what else to say about them. I did some Levels adjustment and a little cropping in CS3. Can't fix the WB.

Also, here's a video of the Double Diatom.





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NikonUser



Joined: 04 Sep 2008
Posts: 2543
Location: southern New Brunswick, Canada

PostPosted: Tue Aug 31, 2010 5:25 pm    Post subject: Reply with quote

The 1st shot is soft but the 2nd and 3rd are very nice. The resolution is good,
what lenses were you using?

Obviously the lighting could be improved, a hot spot in the center; perhaps some diffusion needed.

Also, the images could be improved with some sharpening and cleaning up the background.

Overall, I think the 2nd image is good enough for the "Through the Microscope" gallery.

Congratulations, you have graduated from "Beginners Micro".
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NU.
student of entomology
Quote – Holmes on ‘Entomology’
” I suppose you are an entomologist ? “
” Not quite so ambitious as that, sir. I should like to put my eyes on the individual entitled to that name.
No man can be truly called an entomologist,
sir; the subject is too vast for any single human intelligence to grasp.”
Oliver Wendell Holmes, Sr
The Poet at the Breakfast Table.

Nikon camera, lenses and objectives
Olympus microscope and objectives
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Mitch640



Joined: 15 Aug 2010
Posts: 2137

PostPosted: Wed Sep 01, 2010 5:19 am    Post subject: Reply with quote

Thanks NU. I forgot to ask for recommendations on making my shots better. I am really glad you did. I used a Tucsen USB 10Mp camera in the eyepiece tube of a trinocular scope. Since I have not found any other software to run this camera and capture stills or video with it, I am forced to use the software that came with it. These were all shot with the the 40x lens on the turret of the Amscope.

As for the hotspot. I think it is mechanical. Probably has something to do with the distance the light has to travel in the tube to the sensor. I still need to track that down. But the hotspot is in every still and video, so a hardware issue. What is diffusion and how do I do it?

As for soft or sharp. Again, the software, I think. The live view on my monitor from this camera and run through the software, is definitily low grade. I still have a long way to go. There is an AWB button, and either an auto or manual Exposure slider, and you can adjust the RGB sliders, but they are very flakey. No time was spent developing this software.

What I really need, is the right adapter to use the Canon 1D3 on the trinocular tube. Wink
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rjlittlefield
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Joined: 01 Aug 2006
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PostPosted: Wed Sep 01, 2010 7:26 am    Post subject: Reply with quote

These images are looking good, especially for "first light" from new equipment. I agree you've graduated.

Image #1, I suspect the scale is wrong by quite a bit. The field diameter of a 40X objective at the subject is typically about 0.5 mm = 500 µm. That comes from assuming a field diameter of 20 mm at the eyepiece, then 20 mm divided by 40 is 0.5 mm = 500 µm at the subject. But that length of 11.79 µm that you're showing would imply a field that is about 25 times smaller.

How did you get the scale?

Regarding the hot spot, you might try tweaking the condenser position up and down a bit. Also I notice that it's a lot more intense in #2 and #3 than it is in #1. Why the difference?

The color balance is not too far off, and can be adjusted in Photoshop. Look at the histogram in Colors mode and you'll see that the peaks of red, green, and blue don't line up. Add a levels adjustment layer. In the adjustment layer, select whichever color is farthest left in the histogram, then fly the center and right sliders until that color's peak in the histogram lines up with the rightmost peak. Repeat for the remaining peak. There's a bit of art involved in deciding whether to move the center slider or the right slider or both, but you'll get the hang of it pretty quickly. For your first image, the background goes neutral when the red center slider is at 1.57 and the green center slider is at 1.30 (with the blue center slider left at 1.00 and all right sliders left at 255).

--Rik
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Mitch640



Joined: 15 Aug 2010
Posts: 2137

PostPosted: Wed Sep 01, 2010 7:44 am    Post subject: Reply with quote

Thanks Rik. I am feeling much better about this new hobby already. Now, if I could just get something going with the Canon 1D3 adapter.

I would like to ask here, about getting new objective lenses. The same comapny that I bought the scope from, sells replaceable objectives at a decent price. I think the objectives I have now are just regualr lenses. I see I can replace the 4x and 10x with P A's for about $75. Is that a good price, too good, or should I use another "known" brand? The turret head uses a DIN/JIS standard thread. Wouldn't PA lenses increase image quality?

I got the scale, using the camera supplied software and the etched slide that came with the camera. Then I input the marks from images taken of the slide on all 4 lenses.

I will try the color changes and adjusting the condenser, [although I think I did try that], and get back to you with the outcome.
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Mitch640



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PostPosted: Wed Sep 01, 2010 9:46 am    Post subject: Reply with quote

A followup on the bright spot. Through trial and error, I have eliminated the problem, by removing the 0.5x reducing tube that came with the Tucsen camera. Cheap optics I guess. Here's the scope I bought.. If you click on the image of the scope, you can see the trinocular adapter that came with that scope. It has a screw adjustment to raise it up or down by maybe an inch or so. What isn't shown, is a C-mount adapter that fits over the silver tube at the top. It just so happens, that the thread on that C-mount fit the threads in the body of the Tucsen camera when you unscrew the tube with those cheap lenses in it.

So, putting the C-mount adapter into the camera and that on top of that silver thing, everything works much better, with no bright spot, and maybe some reduction in chromatic abberation that messes with the White Balance. I still need to check that some more.
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NikonUser



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Location: southern New Brunswick, Canada

PostPosted: Wed Sep 01, 2010 10:55 am    Post subject: Reply with quote

If that middle diatom is a single frame from your 40x objective then you are getting great image quality.
I would hold off on buying new lenses but perhaps try stacking several frames using Zerene Stacker. I think you can get if free for 30 days to try.
Once you have perfected lighting (solved already), correcting white balance, possibly some sharpening and cleaning up some of the 'junk' that invariably comes with pond samples then perhaps you could consider a higher quality lens.
I am impressed with the image from that 40x.
_________________
NU.
student of entomology
Quote – Holmes on ‘Entomology’
” I suppose you are an entomologist ? “
” Not quite so ambitious as that, sir. I should like to put my eyes on the individual entitled to that name.
No man can be truly called an entomologist,
sir; the subject is too vast for any single human intelligence to grasp.”
Oliver Wendell Holmes, Sr
The Poet at the Breakfast Table.

Nikon camera, lenses and objectives
Olympus microscope and objectives
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Mitch640



Joined: 15 Aug 2010
Posts: 2137

PostPosted: Wed Sep 01, 2010 11:58 am    Post subject: Reply with quote

That sounds like good advice. Zerene was my next purchase. I admit I never thought of cleaning up the background, but that's good advice too. I use to do it a lot with my bird pics. Simple in CS3.

I admit, I did not know what to expect from the microscope, but I am very happy, for the price I paid. I'd like to get rid of that halo though. Smile
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Charles Krebs



Joined: 01 Aug 2006
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PostPosted: Wed Sep 01, 2010 12:31 pm    Post subject: Reply with quote

Mitch,

I've put together a pdf that covers some basic but important aspects of mounting a camera to a trinocular head. Nothing here that is not disbursed throughout the forum messages, but at least it is now in "one place".

http://www.krebsmicro.com/pdf/trinoc_a.pdf


Also... there's a free program called "Micam" that some people really like for capturing images with eyepiece cameras.
http://science4all.nl/?Microscopy_and_Photography
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Mitch640



Joined: 15 Aug 2010
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PostPosted: Wed Sep 01, 2010 1:40 pm    Post subject: Reply with quote

Thanks Charles. That helps make it more understandable. Just don't try and explain how lenses work with math. You'll lose me quick. I have taken some quick and dirty pics of the trinocular port and the adapter, to help explain what I've got here. I also found all kinds of dust I never saw before. Yikes.

Here is the trinocular port on the nose piece. You can see there is a lens of some kind right inside there. Probably for the sole purpose of collecting dust and sandwich bits. It has a rather large thread in there, that was made to fit the adapter supplied by the scope manufacturer.


And here is the whole thing, put together, after I blew off the dust with canned air. I guess I was lucky that the C-Mount on the top of the adapter had a matching thread to the camera body. This setup also has the added benefit of clearing up the hot spot problem and that bright ring we saw the other day.


But alas, this is not the Canon 1D3 either. I'd love to finagle some kind of adapter to fit that up there correctly.

And thanks for all your help everybody. It does help, and I feel welcome for it. Smile
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rjlittlefield
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Joined: 01 Aug 2006
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PostPosted: Wed Sep 01, 2010 2:28 pm    Post subject: Reply with quote

Mitch640 wrote:
I would like to ask here, about getting new objective lenses. The same comapny that I bought the scope from, sells replaceable objectives at a decent price. I think the objectives I have now are just regualr lenses. I see I can replace the 4x and 10x with P A's for about $75. Is that a good price, too good, or should I use another "known" brand? The turret head uses a DIN/JIS standard thread. Wouldn't PA lenses increase image quality?

I assume that PA means "plan achromat". The part about "plan" means that they have a flatter field than basic achromats. Flat field is very important if you are shooting single frames, not so important if you are stacking for extended depth of field. Plan achromat lenses may also be sharper away from center, but that's not guaranteed. I agree with NU that you should see whether stacking gives you the quality you want, before shelling out for new objectives. $75 is a reasonable price for new minor-brand plan achromat objectives.

Quote:
I got the scale, using the camera supplied software and the etched slide that came with the camera. Then I input the marks from images taken of the slide on all 4 lenses.

I still suspect that something went awry in the process, so that the final number is way off. To put things in perspective, 11.79 microns is only about 20 wavelengths of green light. Granted that image is a little blurred, but it's not nearly as blurred as it would be if you really could fit only 20 cycles of light from end to end.

Can you show us the images of the etched slide, that went into the software?

--Rik
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Mitch640



Joined: 15 Aug 2010
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PostPosted: Wed Sep 01, 2010 2:46 pm    Post subject: Reply with quote

I just deleted those images today. I will take some more in the morning. The software is really low quality and very crashy and I haven't found anything else that will recognize the camera.
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Charles Krebs



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PostPosted: Wed Sep 01, 2010 3:02 pm    Post subject: Reply with quote

Quote:
But alas, this is not the Canon 1D3 either. I'd love to finagle some kind of adapter to fit that up there correctly.


You have a removable trinocular tube. Just for consideration....

1) Remove the trinocular tube.

2) put a high contrast subject on the microscope and focus with the eyepieces

3) Carefully! hold and and lower your Canon body centered over the trinocular tube to see if you can get an in-focus image in the camera. (Be cautious that you don't lower so far that anything will enter into the body mount and possible hit the mirror)

This little exercise is solely to determine whether or not it might be possible to come up with a "direct projection" arrangement. If it is possible it's worth considering.
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rjlittlefield
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PostPosted: Wed Sep 01, 2010 3:22 pm    Post subject: Reply with quote

Regarding scale, there is a simple and reliable method that uses no math and no black-box software.

In Photoshop, just take an uncropped image of your subject and layer it with an uncropped image of your etched slide shot with the same optics. Make one of the layers partly transparent so you can see them both at the same time. Now you have your scale reference laid over your image, as illustrated at http://www.photomacrography.net/forum/viewtopic.php?t=1079. If you want to, you can make a direct measurement just as if you were using a ruler. Or without even making a measurement, you can draw a scale bar using the etched slide as a guide, as shown at http://www.photomacrography.net/forum/viewtopic.php?t=4030.

Other procedures that are more math-based are described in the two "sticky" topics at the top of Beginners Micro, Scale Bars, Part I and Scale Bars, Part II.

I hope this helps. The topic of how to get accurate scale comes up periodically at photomacrography.net, but obviously we don't have it reduced to a clear and concise description yet.

--Rik
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Mitch640



Joined: 15 Aug 2010
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PostPosted: Wed Sep 01, 2010 3:28 pm    Post subject: Reply with quote

What is a removable trinocular tube? That adjustment tube I showed above?

You mean. Tke it off, then take the lens off the the camera and just kind of hold it over the hole and see what I see?

I think I see where your going with this. I already do have a Canon T-ring and an adapter that will clamp onto that silver tube you can see just below the C-Clamp in the picture. Now the problem with this is, with the scope supplied adapter, AND the Canon adapter, it is just way too long. So, the image that hits the sensor is just a tiny part of what's coming through. I need to get the sensor closer, but right now, I have nothing that will match up the threads

But, I will try it and see what I see, and report back. Smile
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