help :\

Starting out in microscopy? Post images and ask questions relating to the microscope and get answers from our more advanced users on the subject.

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mayapapaya
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Location: Fort Lauderdale, FL

help :\

Post by mayapapaya »

Hello hello!

Well after months of obsessively combing through this forum I finally decided it was time to become an official member in an effort to consult those far wiser than I.

Here's the deal....
Everyone around me is a photographer and macro is the cool thing to do. Well I wanted to go a level deeper and try my hand at photomicrography. It's been a really frustrating journey!

I figured a few hours of google and I'd be on my way.... so I did that and made the mistake of purchasing one of those Amscopes. The T490-B, I believe. Promptly returned that sucker. This purchase was made before I even knew what Plan was or that things that should have been movable on this unit weren't.

I purchased a Zeiss Standard yesterday. Looks like either a 14 or 16, though I won't know until I get it in a few days. I'm gonna post pics in a sec. Anyway, now I'm reading all of this crap about infinity corrected optics and stuff. My question/concern is that this might be a really old scope. I'd be using a 2x generic adapter going into where the ocular would sit. I realize that there's the possibility that the eyepieces are built to compensate for whatever little ...aberrations there might be? Is that right? I'm so confused. What about the body? even if I removed the binoculars from the path and abandoned the adapter altogether would there still be some sort of compensating glass in the way? Maybe I should just post what I want to be able to do and someone can tell me how I can facilitate them:

1. Taking the actual picture: Better to use adapter or just sit it on top? I have an extension tube, I don't know if this makes a difference or if it should be implemented.

2. I want to do dark field. Do I really need a different condenser designated for it or will those nifty little stoppers work?
Is it only the condenser that needs modifying? What about the objectives? Can regular BF objectives be used for DF?

3. What about phase contrast? Same deal? Is there a diy for this? Special objectives? Sigh.

4. How about lighting? Now I think this is a 15W illuminator. Can I add more light with some mini goosenecks or something positioned under the specimen? How about those fluar objectives (looks like mine came with one). Do I need special lighting for that? Something I could rig myself?

5. What the heck is DIC? I read all this stuff about how expensive it is and how great it is but Differential Blah Blah doesn't mean anything to me. Anyone care to explain it in a simplified manner?

6. Lastly, take a look at this thing. What will I be able to achieve with it as is (from what you can see)? Worth keeping or sending back? Cost me $360. What do you think? How can I beef this up (if it's a solid starting point)? What can I DIY and what needs to be purchased?

PLEASE HELP.

I'd be ever so grateful. I know I've probably exceeded my question limit :P Help this lovely lady out!




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rjlittlefield
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Re: help :\

Post by rjlittlefield »

Maya, welcome aboard! :D

I will let other more experienced people address your questions specifically about the Zeiss unit. But perhaps I can help with some of your more general questions.
2. I want to do dark field. Do I really need a different condenser designated for it or will those nifty little stoppers work?
Is it only the condenser that needs modifying? What about the objectives? Can regular BF objectives be used for DF?
A simple stop should be fine to start. You can use BF objectives. Except at very high magnification, there's nothing special about objectives for DF. (At high mag, objectives for DF may require a smaller aperture than for BF, to allow the condenser to illuminate the specimen without shining directly into the objective.)
3. What about phase contrast? Same deal? Is there a diy for this? Special objectives?
No, phase contrast is completely different and there's no DIY. Both the objectives and the condensers are special.
4. How about lighting? Now I think this is a 15W illuminator. Can I add more light with some mini goosenecks or something positioned under the specimen? How about those fluar objectives (looks like mine came with one). Do I need special lighting for that? Something I could rig myself?
I'm guessing that you'll need more light only for photography, to keep the exposure time down. In that case you can probably do a DIY trick of placing a piece of glass under the condenser, slanted at 45 degrees to reflect light from an electronic flash mounted off to the side, up into the condenser. That will let you freeze fast-moving protists and the like. The same trick does not work very well with continuous light sources because the efficiency is low compared to what's built into your microscope. With the builtin, most of the light gets delivered up through the condenser; with the glass reflector, most of the added light will get lost.
5. What the heck is DIC? I read all this stuff about how expensive it is and how great it is but Differential Blah Blah doesn't mean anything to me. Anyone care to explain it in a simplified manner?
Imagine that you're looking at a piece of glass that is clear but has uneven thickness. Ordinary brightfield is like looking at the glass with a uniform big white cloud behind it -- the differences in thickness are barely visible. Phase contrast turns differences in thickness into differences in brightness, at the cost of introducing characteristic halos that greatly change the overall impression. DIC also turns differences in thickness into differences in brightness, but in a different way that does not introduce halos and produces a fairly "natural looking impression" of 3D. The quotes are there for a reason: the impression of 3D produced by DIC doesn't always match the real 3D structure of the subject. But it does make the structure visible and the images look great.

There are other and much less expensive ways of making the structure visible. They have names like "oblique lighting" and they're essentially similar to brightfield except that the uniform white cloud gets replaced by one that has some areas darker than others. This allows small differences in thickness to produce obvious differences in brightness, rather like how very shallow ripples on the surface of a swimming pool can produce obvious "shadows" and bright spots on the bottom of the pool by redirecting the light from the sun. Such lighting can be produced with a normal brightfield condenser by sticking various filters underneath it. In the forum here, see THIS TOPIC and search for other topics with universal AND filter by username litonotus. He is the master of the art.

I use the word "art" deliberately. One big reason DIC is much loved by researchers is that it's very repeatable, unlike oblique lighting and related techniques where different users and different scopes are likely to produce much different and often inferior results.

Again, welcome aboard. I'll be away from the forum for a couple of days, but hopefully other people will chime in to give more info & advice.

Cheers,
---Rik

mayapapaya
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Post by mayapapaya »

Thank you kindly for taking the time to answer my questions in such an easily digestible way. Much appreciated :)

Pau
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Post by Pau »

mayapapaya wrote:I purchased a Zeiss Standard yesterday. Looks like either a 14 or 16,
It's a 14, the only difference is that the 16 has 5 objective turret, but it seems the version that has built in 10W 6V hallogen lamp and transformer.
Yes, it's old but a very good one, I have two and love them. Not at the level of modern infinite research scopes from the big makers but IMO better than the modern cheap finites from China or India. One advantage of this model is the high availability of parts in the used market as those series where made during a long period and many parts are compatible with the big ones like Universal and Photomicroscopes.
mayapapaya wrote:I realize that there's the possibility that the eyepieces are built to compensate for whatever little ...aberrations there might be? Is that right?
Yes, and not little ones!. For good quality the only way is to shot trough original Zeiss KPL eyepieces, and the simpler and more convenient form will be the called afocal method. I did a post about this topic some time ago:
http://www.photomacrography.net/forum/v ... hp?t=15607
Because your head is not trinocular you would neet to set up an adapter at the right angle over one of the eyepieces, not that convenient but workable.
mayapapaya wrote:I want to do dark field. Do I really need a different condenser designated for it or will those nifty little stoppers work?
Is it only the condenser that needs modifying? What about the objectives? Can regular BF objectives be used for DF?
The Rik's answer is right, but with the normal Zeiss condenser is difficult to place a darkfield stop in the right position (near the condenser diphragm). It's easier with a turret phase condenser. There are also dedicated DF condensers for objectives of higher NA
mayapapaya wrote:What about phase contrast?
You need dedicated and matched Phase condenser and objectives. Not difficult to find used at reasonable (but not very cheap) prices
mayapapaya wrote:What the heck is DIC?
Forget about DIC, at least until you was a more experimented microscopist. It is VERY expensive and difficult to find

The price you paid seems very reasonable if it is in good working condition. In the future you would want to buy many parts like better objectives, a trinocular head, condensers, may be polarizers....so it is difficult to give advice. Just to say that my own experience with the Zeiss standard is very satisfactory.

But my main advice is for you is to learn both reading and practicing microscopy, even without taking pictures from the beguining.
Some links of interest:

A very good introduction to microscopy from Zeiss:
http://www.zeiss.de/C1256B5E0051569F/Em ... inning.pdf

More advanced topics (like DIC...):

http://www.olympusmicro.com/primer/tech ... index.html

http://micro.magnet.fsu.edu/primer/index.html

To download manuals of Zeiss microscopes:
http://www.science-info.net/docs/zeiss/
Pau

mayapapaya
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Location: Fort Lauderdale, FL

Post by mayapapaya »

very good, all useful information! thank you very much. yeah, i do have a tendency to put the cart before the horse, as they say. so Pau, you're suggesting to abandon that adapter I have altogether since it won't correct what I'm seeing? I've actually never read up on photographing through the actual eyepieces.... i'll have to check out that link. about getting better objectives, i'm wondering what would be compatible? from what i've read there are different working distances and even threads for different manufacturers and I also read that it was better to stick with the brand of microscope due to it's tube length. what are my options there? are there devices to compensate for any slight differences in distance?

Pau
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Post by Pau »

The Z. Standard is a DIN microscope, so you need objectives corrected for 160mm tube lengh. You may mount any DIN objective (I mostly use Leitz ones), but the aberration corrections are not standardized, so, if you already have Zeiss objectives and eyepieces the easier way may be to remain with Zeiss, but if you find a good deal in other high quality mayor brand you may switch.

Of the three objectives you showed the 40X Neofluar is a very good one, the other two just seem achromats but modern ones.
From worse to better the objectives are classified:
-about flatness of field
(nothing marked)--->Semiplan--->Plan
-About chromatic correction
Achromat (usually nothing marked)--->Fluorite---->Apochromat

I most use Plan fluorites and few Fluorites and plan apos, and all of them are very good.

So the worst, in special for photomicrography, are Achromats and the best ones and MUCH more expensive PlanApos.
The Neofluar are fluorites with decent flatness of field, I would qualify them as semiplan. For a 40X this would be enough flat with most specimens.

About the adapter if you don't have the adequate correction only the very center of the image will be good, with severe CA to the periphery.

Nikon made DIN objectives that doesn't need corrections at the eyepiece (called CF), very popular in the forum to shot bellows mounted, and some modern oriental ones also seem to do not need correction, but the info about them is scarce and insure.

But again, excuse me to repeat, first learn to use the microscope and later plan the equipement you need :D
Pau

mayapapaya
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Post by mayapapaya »

yeah, i have some lab experience as it is from university and such but just the basics. just want to be prepared and knowledgeable for when i'm ready to move forward :) thanks again, Pau.

Mitch640
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Post by Mitch640 »

"What about the body? even if I removed the binoculars from the path and abandoned the adapter altogether would there still be some sort of compensating glass in the way?"

I did not see this answered. I do not know about your scope, but take the binocular head off and look. I use a Nikon Fluophot and it has a lens in the arm between head and turret. It's a nasty trick Nikon played on people and it's an absolute bear to clean, which needs doing often. I also have an Olympus BH-2 base, no head, and it also has a lens in the end of the arm between head and turret. So that's a 100% hit ratio, 2 for 2.

Pau
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Post by Pau »

Mitch640 wrote:"What about the body? even if I removed the binoculars from the path and abandoned the adapter altogether would there still be some sort of compensating glass in the way?"

I did not see this answered. I do not know about your scope, but take the binocular head off and look. I use a Nikon Fluophot and it has a lens in the arm between head and turret. It's a nasty trick Nikon played on people and it's an absolute bear to clean, which needs doing often. I also have an Olympus BH-2 base, no head, and it also has a lens in the end of the arm between head and turret. So that's a 100% hit ratio, 2 for 2.
Mitch, I think I answered about the need to place a compensating KPL eyepiece.
In the Zeiss Standard there isn't any lens between the objective and the microscope head in the basic configuration, and in more complex setups with telan lenses to put intermediate optics (like mine) all the compensation is still done at the eyepiece.
Pau

Alan Wood
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Post by Alan Wood »

Mitch640 wrote:I also have an Olympus BH-2 base, no head, and it also has a lens in the end of the arm between head and turret.
You must have a BHTU stand. The BHT and BHS stands do not have any glass between the nosepiece and the head.

Alan Wood

Mitch640
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Post by Mitch640 »

Alan, yes, a BHTU.

mayapapaya
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the results are in...

Post by mayapapaya »

well folks, with what i currently have available to me this is the result i got: http://www.photomacrography.net/forum/v ... 095#114095

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