Help with decision on Leitz Orthoplan

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Alexander
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Re: Help with decision on Leitz Orthoplan

#31 Post by Alexander » Sun Jan 29, 2023 1:34 am

InfiniteCosmos wrote:
Sat Jan 28, 2023 9:57 pm

This ploemopak works fine with infinity objectives?
Because they seems to be way cheaper and abundant.
The Ploemopak is designed to work with standard transient light objectives made for finite tube length 170 or 160 mm. Infinity objectives will not work.
To make it worse: In reflected light fluorescence work the objective is the condenser as well. To achieve good fluorescence you need objectives as powerful as possible. A high NA is crucial. 10/0.45, 20/0.75 and 1.3/40 are typical objectives used for this application. Your powerful lightning will help of course.
Leitzcycler wrote:
Sat Jan 28, 2023 8:07 pm
Be careful with the mercury lamp as I think it may be even more dangerous if used with transient light.
Very much! It is quite dangerous to use a HBO 250 in transient light. A Ploemopak is a much safer configuration.

viktor j nilsson
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Re: Help with decision on Leitz Orthoplan

#32 Post by viktor j nilsson » Sun Jan 29, 2023 5:05 am

InfiniteCosmos wrote: Okay so i think im getting it. The 160mm and 170mm are both the same phisical size and i will not have to make any ajustment on the stage when changing between these two objectives. The only change is were the image they produce is being delivered, in the eyepiece-hole of the head.
Is that it? Am i finally learning the basics? :lol:
Yes, and the difference in where the image is produced is very small: 152mm vs 150mm above the shoulder of the objective. The big difference lies in the eyepieces, which is why it's so important to use 170mm eyepieces on 170mm scopes.

But note that there also existed an earlier series with "short" 170mm objectives, having a ~37mm parfocal distance. You probably don't won't to mix those with your "long" 45mm objectives.

InfiniteCosmos
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Re: Help with decision on Leitz Orthoplan

#33 Post by InfiniteCosmos » Sun Jan 29, 2023 10:27 am

I saw a ploemopak with infinity objectives somewere and i though that could be possible. Maybe i saw something out of context or for other similar Letz series.
So this Ploemopak for sale can be used with epi-fluerescence too if i later make the transition for LED? Since is much safer.

A part of me wants to start from the basics and focus on the Darkfield condenser and the iris objective like imkap said, the other part of me is saying this ploemopak can be a good investment at a good price since it is the missing part i need to complete my fluorescence kit. And also the same Silver Grey color too.
A third part of me got alot of read and understanding to do :lol:

Oh okay, those high NA objectives work as good for non-fluorescent microscopy too? Because I was here searching for Plan objectives and maybe i can buy objectives for both regular lighting and Fluorescent lighting. Saving me some money that way.

Great awareness about those 37mm parfoacal objectives. I could easily made a mistake there.

Leitzcycler
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Re: Help with decision on Leitz Orthoplan

#34 Post by Leitzcycler » Sun Jan 29, 2023 12:53 pm

So this Ploemopak for sale can be used with epi-fluerescence too if i later make the transition for LED?
Yes. If you are seriously interested in fluorescence microscopy, that Ploemopak for affordable price would be a very good investment. Personally I have no experience with transmitted fluorescence. However, it is not used much these days.
Oh okay, those high NA objectives work as good for non-fluorescent microscopy too?
Yes. But now we are talking mostly at professional level I think. With ploemopak and leds, I think you may get reasonably good results with any objective at hobby level. I would first buy the ploemopak, construct the led illuminator and then test it with objectives you have. Then you may make investments for more specialized objectives if you decide to continue. They may be very expensive.

Hobbyst46
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Re: Help with decision on Leitz Orthoplan

#35 Post by Hobbyst46 » Sun Jan 29, 2023 1:06 pm

The main problem with transmitted light fluorescence, which made it outdated long ago, is that it is extremely wasteful of illumination. Fluorescence requires both intense illumination for excitation AND efficient removal of that exciting light such that the fluorescence sensor (eye, camera, whatever) does not see the excitation. Meeting both of these requirements is much more successful via epi-illumination than via transmitted illumination.

Leitzcycler
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Re: Help with decision on Leitz Orthoplan

#36 Post by Leitzcycler » Sun Jan 29, 2023 1:15 pm

I don't know if you Phaco objectives would work well. I would guess the phaco ring would do some harm for the light intensity. Not sure what will happen, never tried myself.

MicroBob
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Re: Help with decision on Leitz Orthoplan

#37 Post by MicroBob » Sun Jan 29, 2023 2:58 pm

Hobbyst46 wrote:
Sun Jan 29, 2023 1:06 pm
The main problem with transmitted light fluorescence, which made it outdated long ago, is that it is extremely wasteful of illumination. Fluorescence requires both intense illumination for excitation AND efficient removal of that exciting light such that the fluorescence sensor (eye, camera, whatever) does not see the excitation. Meeting both of these requirements is much more successful via epi-illumination than via transmitted illumination.
I can remember another disadvantage being mentioned: The fluorescence takes place at the underside of the object which leads to less clarity in the image. Can you comment on this, Doron?

Hobbyst46
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Re: Help with decision on Leitz Orthoplan

#38 Post by Hobbyst46 » Sun Jan 29, 2023 3:59 pm

MicroBob wrote:
Sun Jan 29, 2023 2:58 pm
Hobbyst46 wrote:
Sun Jan 29, 2023 1:06 pm
The main problem with transmitted light fluorescence, which made it outdated long ago, is that it is extremely wasteful of illumination. Fluorescence requires both intense illumination for excitation AND efficient removal of that exciting light such that the fluorescence sensor (eye, camera, whatever) does not see the excitation. Meeting both of these requirements is much more successful via epi-illumination than via transmitted illumination.
I can remember another disadvantage being mentioned: The fluorescence takes place at the underside of the object which leads to less clarity in the image. Can you comment on this, Doron?
I am not sure about this, Bob, but am guessing as follows.
The transmitted excitation light beam hits the specimen sideways, at a nearly right angle, for example, by using a darkfield condenser or a darkfield stop. So the illumination excites fluorescence from all surfaces of the specimen. If the specimen is thin enough, and transparent, fluorescence emitted from the bottom of the sample can still pass and be detected. If the specimen is thick and/or highly absorbing (for example - an overly-stained tissue), transmitted fluorescence will be poor, except from the portion from the top surface of the specimen. In that case indeed, epi-fluorescence will be advantageous (in addition to the other reasons).

BTW, in fluorometers, instruments that enable the sensing and measurement of fluorescence from a solid or liquid specimen,
the configuration is the same: The excitation source, specimen and sensor occupy the vertices of a right-angle triangle.

P.S. even under epi-illumination, unwanted out-of-focus contribution to the fluorescence image (haze) were one of the reasons (or the chief reason - not sure) why confocal microscopy has been developed...

InfiniteCosmos
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Re: Help with decision on Leitz Orthoplan

#39 Post by InfiniteCosmos » Tue Jan 31, 2023 11:45 am

Hi there friends,

Since i lost the Ploemopak auction, sadly, i will now focussing on getting decent objectives for my orthoplan having in mind that for now i want to achieve Darkfield and later will be using fluorescent or/and epi Fluorescent lighting.

Sorry for all the confusion that comes next:
For now i would like to be able to do darkfield on 40x/63x/100x objectives. All this objectives have to have an iris on them to achieve Darkfield? Should i search for the high NA objectives too?
So as the condensers, i saw one describing as a darkfield but only with 0.80-0.95 value, and i though the condenser had to be a 1.20. So im a bit confused here.
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When people talk about cheap darkfied achieved with filter stops, thats only for magnifications lower than 40x because of the NA of the objective, right?

PS: Im assuming this, please correct me if im wrong:
Transmited Light: light source -> condenser -> sample -> objective -> eye.
Transient Light: light source -> objective -> sample -> objective again -> eye
Last edited by InfiniteCosmos on Tue Jan 31, 2023 12:20 pm, edited 1 time in total.

Alexander
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Re: Help with decision on Leitz Orthoplan

#40 Post by Alexander » Tue Jan 31, 2023 11:55 am

The two pictures show two different dark field condensers. The first is a dry one suitable for objectives up to NA of about 0.7. The second one is an oil immersion condenser suitable for objectives with NA up to about 1.0. The handling of this condenser is somewhat tricky as well. Even with the latter one you need 63x or 100x objectives with iris. Those are quite rare, even rarer when supporting the large field of view featured by the Orthoplan.

Darkfield works best with medium NA objectives.

Leitzcycler
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Re: Help with decision on Leitz Orthoplan

#41 Post by Leitzcycler » Tue Jan 31, 2023 1:06 pm

Since i lost the Ploemopak auction, sadly,
I am so sorry. It went for a good price. Don't worry, they will come. :)

MichaelG.
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Re: Help with decision on Leitz Orthoplan

#42 Post by MichaelG. » Tue Jan 31, 2023 2:55 pm

InfiniteCosmos wrote:
Tue Jan 31, 2023 11:45 am
So as the condensers, i saw one describing as a darkfield but only with 0.80-0.95 value, and i though the condenser had to be a 1.20. So im a bit confused here.
.

This is the more modern version of the leaflet that I posted recently:
http://www.science-info.net/docs/leitz/ ... denser.pdf

Essential reading, I would suggest.

MichaelG.
Too many 'projects'

apochronaut
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Re: Help with decision on Leitz Orthoplan

#43 Post by apochronaut » Tue Jan 31, 2023 9:11 pm

Alexander wrote:
Tue Jan 31, 2023 11:55 am
Darkfield works best with medium NA objectives.
Darkfield works easiest with medium N.A. objectives. It works best with high N.A. objectives.

InfiniteCosmos
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Re: Help with decision on Leitz Orthoplan

#44 Post by InfiniteCosmos » Thu Feb 23, 2023 10:00 pm

Hi there friends,

Thank for all the advice you guys gave me this last month.
Thanks for the Leaflet too Michael.

Ive been keep searching and reading about the wild phototube adaptations i have to make and searching for the condensers and the objectives that i want to buy. When i have my setup complete i will post something on "My Microscope" section.
Meanwhile, i got two Plan objectives and i now able to properly use the microscope :D
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The light bulb died 1 week right after i got the microscope. Change the bulb sounds pretty easy until you find out that the bulb have no screw thread at all :lol:
I end up removing the light casing and just fit a regular 9w 5000k led bulb right inside of the microscope. It sounds crazy i know, but it seems to work pretty better than the previous one.
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I Have a question :x
I think you guys may have some experience with different types of objectives that i dont.

I wonder if a 40x PLAN APO with 0.75 NA, is actualy better that a 40x FLUOTAR with 1.30 NA.
As far as i know, the PL APOs are better corrected colour aberration than the Fluotars, i think this means more resolution too, am i right?.. Since there is less aberrations the image should be more sharp.
But also, a High NA such as 1.30 on a 40x Fluotar objective its a huge diference compared to a 40x Plan Apo 0.75. I have been reading about the importance of the NA to the image resolution.

Can you guys debug my mind over this?
Have some of you came across this scenario?

Alexander
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Re: Help with decision on Leitz Orthoplan

#45 Post by Alexander » Thu Feb 23, 2023 10:47 pm

Don't compare apples with pears.

The PL Apo 40/0.75 is a dry objective while the PL Fluotar 40/1.3 is an Oil one. Resolution is a function of NA. The 40/1.3 provides significant higher resolution than the 40/0.75.

A PL Apo is perfectly corrected against chromatic aberrations while the Fluotar is corrected only for 2 of the 3 colors. In real life the difference is negligible. The Fluotars deliver a superb picture.

In Fluorescence work chromatic aberrations are a no-brainer anyway. I would choose the Fluotar over the PL Apo without any hesitation.

Hobbyst46
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Re: Help with decision on Leitz Orthoplan

#46 Post by Hobbyst46 » Thu Feb 23, 2023 10:53 pm

A 40X/1.30 objective is most useful for fluorescence microscopy, since its high NA collects a lot of fluorescence from the specimen. This is important because fluorescence in general is weak, so, the more light gathered, the better. The high NA also means high resolution, as you mentioned.
On the other hand, that objective must be used with oil immersion, and in addition, its working distance is very shallow.
So, for general purpose hobby microscopy (if there is such an animal), it is less useful than the 40x/0.65 or 40x/0.75 planachromats/fluotars, say.

P.S. as I was typing I noticed Alexander's response. Sorry for any redundancy.

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woyjwjl
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Re: Help with decision on Leitz Orthoplan

#47 Post by woyjwjl » Fri Feb 24, 2023 8:56 am

Hello, I happen to have an Olympus BHSP for sale at a price of less than $500 (excluding polarization kit). The quality and scalability of BH2 need no more introduction. If you are interested, please contact me via private message.



Best wishes, Lei
Micrographers from China, thanks to the forum for providing a platform for exchange

InfiniteCosmos
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Re: Help with decision on Leitz Orthoplan

#48 Post by InfiniteCosmos » Fri Feb 24, 2023 1:37 pm

Great information!

I was thinking here... if the Fluotar 40x/1.30 objective lacks the iris i will never be able to perform a Darkfield effect with that objective. Right?

Alexander
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Re: Help with decision on Leitz Orthoplan

#49 Post by Alexander » Fri Feb 24, 2023 1:41 pm

Right. No Darkfield with the Fluotar 40/1.30.

InfiniteCosmos
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Re: Help with decision on Leitz Orthoplan

#50 Post by InfiniteCosmos » Wed Mar 01, 2023 11:53 am

Hi again!
A friend o mine have what i think to be a part for epi-illumination for orthoplan. A part that i dont have on my microscope. And since i have the goal of making Fluorescence microscopy...
Ive tried to find an Orthoplan parts list on internet but i couldnt find.
I search on the manuals you guys gave me and i found nothing on fluorescence parts.
Yesterday i saw somewone selling some orthoplan stuff on ebay while having this image on the selling page:
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Its a parts list that i wish to have. Anyone knows were can i find this manual?


The part my friend want to sell me is this one:
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He says this part goes right between the ploemopack tube and the epi-illumination hole of the microscope.

Is this part necessary for epi-illumination and epi-fluorescence?
He also says this part have a diffusion filter named(Diffus N) that i dont know what is for.

Leitzcycler
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Re: Help with decision on Leitz Orthoplan

#51 Post by Leitzcycler » Wed Mar 01, 2023 8:00 pm

I have just a simple tube. In your tube in the picture there is probably an iris diaphragm. Maybe useful in some applications. Ploemopak has its own diaphragm. The tube will prevent light scattering. The diffusion filter makes the lightning more even and important with many more primitive lightning systems. Not necessarily needed with Orthoplan I think.

I have instruction for Ploemopak in PDF. If you need it, please send your email to me via personal message and I will send it to you.

Leitzcycler
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Re: Help with decision on Leitz Orthoplan

#52 Post by Leitzcycler » Wed Mar 01, 2023 8:14 pm

The tube (in your picture) might be originally for another type of epi illuminator nosepiece. My tube is 76,5mm of total lenght. If the length is ok you may use it with ploemopak.

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