I need some help...

Images made through a microscope. All subject types.

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Mr.Stone
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I need some help...

Post by Mr.Stone »

Good evening all :)

I got stuck this weekend trying to figure out how to take pictures through my wild m20. But no matter what i tried i just dont get clear pictures, colours are good but the clarity aint great at all.
I first tried just taking pictures with the camera connected to the microscope, and the 6x photo eyepiece. Then tried putting the camera on bellows so that there would be no shake, but this made no difference.
Then i fitted a teleconveter onto the camera then the camera to the microscope, so the image passed through the 6x photo eyepiece, then through the converter then onto the sensor.... the images were a bit better but still not as clear as they should be.
So im out of ideas as to what else i can try to make things look better.

Here are some pictures i took, im not too happy posting these as i think that they look baaaad, but i need your help :)
1
Image
2
Image
3
Image
4
Image
5
Image

The top 4 pictures were taken with a 10x phase 0.45 na objective and the last pic is taken with a 40x phase 0.75 na objective.
I did not use flash here, and i used phase contrast method to light up the subjects. These are how the pictures came out, no editing or sharpening was done, i just took out some sensor dust.
These all were taken on the same slide.
I used the two second delay on my camera, this lifts the mirror, then there is a 2 second pause, then a smaller shutter opens and exposes the image closes then the mirror drops.

How can i get sharper pictures with this setup, or is this how good it will get?

Charles Krebs
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Post by Charles Krebs »

I used the two second delay on my camera, this lifts the mirror, then there is a 2 second pause, then a smaller shutter opens and exposes the image closes then the mirror drops.
What camera?
I first tried just taking pictures with the camera connected to the microscope, and the 6x photo eyepiece
How did you configure this?
Then tried putting the camera on bellows so that there would be no shake, but this made no difference.
Again, how was this configured?

Might be best to post a few shots of the set-ups over in the"technique" or Equipment " sections. It is hard to make any suggestions without a clear idea of the way this was arranged.

Mr.Stone
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Post by Mr.Stone »

My apologies Charles.

The camera is a sony a580.
Here are pictures i took this morning before going to work, i was in a rush so i took them with a cellphone.

Image

Image

I used these tubes as can be seen in the pictures above, first time around i connected the camera to the mount at the back (where the white cap is) and then placed this on the silver part of the wild. So the camera is "balancing" on this silver part. It is stable and does not wobble. The silver part can be loosened and adjusted up and down, i tried a variety of heights and none worked.
I then put the 2x teleconverter on the camera and used the same method as above to connect the camera to the microscope but this didnt work either. I know im doing something wrong but im not sure what it would be.
Is it possible to move this post to the right place?
Thank you :)

Olympusman
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Picture clarity

Post by Olympusman »

I recommend rotating the phase condenser to the white light position when you are doing sharpness tests. The haloes created by a phase setup will not give you sharp enough images to judge sharpness. Also, adjust your condenser position so that you have Koehler illumination.
Michael Reese Much FRMS EMS Bethlehem, Pennsylvania, USA

Charles Krebs
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Post by Charles Krebs »

Mr Stone,

Still lots of questions! :wink:
(Here's one needed for background info... Are you using 10X viewing eyepieces?)

Michael is correct. If you are setting up and trying to trouble-shoot a camera on a microscope doing so with phase contrast and relatively thick subjects is not a good idea.

Try something along these lines...

Remove the 6X eyepiece and camera adapter from the trinocular tube.

Find a high contrast, thin, flat, non-moving subject subject. Ideally one that fills the entire view. Use brightfield illumination. Set up the microscope properly (bulb and condenser properly aligned). Be sure the condenser is at proper height and the condenser aperture only slightly closed down (75%-80% of the objective rear aperture is "clear" as viewed through an empty eyepiece tube). Use a lower power objective...10X is good. Focus carefully on the subject. (Be sure to have properly set your interpupillary distance and have the diopter adjustment on the left eyepiece tube set for your eyesight).

Now... without changing the microscope focus, remove the viewing eyepiece from the left binocular eyepiece tube and insert it into the trinocular tube. Look down through the trinocular tube through that eyepiece and see if the subject looks to be about the same size and is in focus (or very close to being in focus). It should be. If the focus is far off then something is wrong. If it is very close then fine tune the microscope focus to bring it into sharp focus as seen through the eyepiece in the trinocular tube.

Next... remove the viewing eyepiece from the trinocular tube. (Don't change microscope focus!) Now insert the 6X Photo eyepiece you have pictured above into the trinocular tube. Look down through that 6X eyepiece and see if the subject is in focus. Let me know what you find.

(Here is one thing I am trying to determine. I am not familiar with the Wild 6X Photo eyepiece you show. There were two basic types of "photoeyepieces" made for the microscopes of this era. One was a "projective" type, and was meant to be used by itself to project an image directly onto the film (or Polaroid) with usually no additional intervening optics. The other type of photoeyepiece was meant to be used with additional optics placed before the film (or Polaroid). This then made up what is known as an "afocal" arrangement. If you get a visually in-focus image through the 6X after inserting it into the trinocular tube then it is the latter type... meant to be used with additional optics. If the view is not in focus then it is likely a "projective" type photoeyepiece, intended to be used with no additional optics. Perhaps someone reading this is familiar with the Wild M20 optics and can offer more information).

Your camera sensor measures about 23.6x15.7mm. If you are using 10X viewing eyepieces, then ideally, the magnification provided by the optics in the trinocular tube should be about 1.5X. If the 6X eyepiece you are using is a projection type, and it is set up properly, it will provide a 6X magnification onto the sensor. This is far too much. It will result in you recording only a small section of what is seen through the viewing eyepieces. A 6X magnification onto an APS-C sized sensor will result in a great amount of what is called "empty magnification".... the results will not look very sharp at all.

(At this point it is important to realize that by changing the microscope focus and/or changing the"proper" (designed for) height/position of the optics and camera in the trinocular you can "force" the trinocular tube magnification to vary widely from what is marked on those optics. You can get an image into the camera using these optics in ways never intended by the designers. This is not a good approach. These optics were designed to be used at very specific locations and focus distances. In some cases it may work out OK, but often the end result will be significantly compromised image quality).

Try the above actions and let us know what you find out. Then we can provide some concrete suggestions.

If you haven't seen it already, you might also take a look at this PDF:
http://www.krebsmicro.com/pdf/trinoc_a3.pdf

Charlie

Mr.Stone
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Post by Mr.Stone »

Still lots of questions!
(Here's one needed for background info... Are you using 10X viewing eyepieces?)
:) Ask away! i have quite alot of questions too.
About the 10x eyepieces, yes, im using them, with the microscope i got 2 6x eyepieces along with a wild centering telescope and a bausch and lomb 12.5x eyepiece(not quite sure what this is for).
Michael is correct. If you are setting up and trying to trouble-shoot a camera on a microscope doing so with phase contrast and relatively thick subjects is not a good idea.
:roll: Yeahhhh, should have thought about this one. I just got a bit too excited about taking pictures through the wild :lol:
(Here is one thing I am trying to determine. I am not familiar with the Wild 6X Photo eyepiece you show. There were two basic types of "photoeyepieces" made for the microscopes of this era. One was a "projective" type, and was meant to be used by itself to project an image directly onto the film (or Polaroid) with usually no additional intervening optics. The other type of photoeyepiece was meant to be used with additional optics placed before the film (or Polaroid). This then made up what is known as an "afocal" arrangement. If you get a visually in-focus image through the 6X after inserting it into the trinocular tube then it is the latter type... meant to be used with additional optics. If the view is not in focus then it is likely a "projective" type photoeyepiece, intended to be used with no additional optics. Perhaps someone reading this is familiar with the Wild M20 optics and can offer more information).
I have taken a look through the 6x photo eyepiece while specimens were in focus, if i recall correctly the image was in focus, but i had to hold my eye a bit further then with the 10x eyepieces. But i will do this again after carefully following all the steps that you mention here and report back to you.

Thank you both Charles, Olympusman for the help :)
I will do this tonight or tomorrow night and let you know how things went.

Mr.Stone
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Post by Mr.Stone »

Charles,
Next... remove the viewing eyepiece from the trinocular tube. (Don't change microscope focus!) Now insert the 6X Photo eyepiece you have pictured above into the trinocular tube. Look down through that 6X eyepiece and see if the subject is in focus. Let me know what you find.
upon placing the 6x photo eyepiece in the trinocular tube the image was sharp and in focus.
So this would be, going on by what you wrote, an "afocal" arrangement that would need some additional optics?

Pau
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Post by Pau »

About afocal:
http://www.photomacrography.net/forum/v ... 9265#99265
and search at the forum for afocal
Pau

Mr.Stone
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Post by Mr.Stone »

Thank you Pau:)

I have a minolta 50mm f1.4 or f1.8 i used to use for reverse this is the old mount and does not fit the alpha body, and then a sony 50mm f2.8, there is also a 24mm f2.8 somewhere, but by the looks of things lenses in the 50mm range seem to do the thing.

Charles Krebs
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Post by Charles Krebs »

I have a minolta 50mm f1.4 or f1.8 i used to use for reverse this is the old mount and does not fit the alpha body, and then a sony 50mm f2.8, there is also a 24mm f2.8 somewhere, but by the looks of things lenses in the 50mm range seem to do the thing.
With the sensor size in your camera and a 6X eyepiece in the trinocular tube, you should try the Sony 50mm lens. (If you run the "numbers" a 60-65mm lens would be just right, but you're not going to find one. With a 50mm you might get a slight amount of vignetting in the corners).

Mr.Stone
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Post by Mr.Stone »

Thank you Charles :)

The closest lens i have to a 60mm is my sony 70-400g lens. But i really doubt this would work, ill try and get the 50mm onto the m20.
The thing that i dont like about the m20 is that when you ajust the focus, the whole "neck" moves instead of just the stage.

Pau
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Post by Pau »

Also try your 50mm lens with a 10X eyepiece. I tend to think that it will give you better coverage at least with some optics, if both eyepieces work, you will have alternate magnifications

6 X 50 / 250 = 1.2X As Charles says likely you will get some vignette

10 X 50 / 250 = 2X, a bit too cropped, but very useful

Of course a 8X high eyepoint eyepiece would be the best option (1.6X )

Forget about the long zoom.
Pau

Mr.Stone
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Post by Mr.Stone »

I will try this thank you Pau.
I have a 8x leitz eyepiece, i cant remember if ist a high eyepoint one tho.
I also have a couple of zeiss ans nikon "bits" that look like they are some sort of camera too microscope adapter, the one has a straight tube with what looks like an old minolta mount at the one end, but it looks slightly smaller then the old minolta mount that im familiar with, then it has an eyepiece like thing that sticks out the side with wires in it that creates a grid similar to the crosshairs on a rifle scope.
When i get the time i will post some pictures of these.

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