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Nikon E600 microscope setup/testing

 
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Jesse



Joined: 11 Dec 2016
Posts: 64

PostPosted: Thu Aug 23, 2018 8:20 am    Post subject: Nikon E600 microscope setup/testing Reply with quote

Ok, I've taken delivery and I've assembled the scope. I cleaned all exposed glass surfaces to the best of my ability with a standard lens cleaning cloth before assembly. To my untrained eye, everything appears undamaged. Things I've noticed immediately:


    The first thing I noticed is that the objectives are quite a bit larger than I expected. I've got a stupid grin plastered on my face. This thing is serious.

    Dust. There's some dust on the mirror of the trinoc port and I see dust pretty much everywhere in the light delivery system. Not sure how much dust is between the objectives and the eyepieces though. Seems cleaner there. I plan to attempt to use a balloon to remove the dust on the trinoc mirror. Should I care about the light delivery system dust at all?

    The illuminator light is very yellow. Is this normal? I don't know why, but I was expecting a whiter light. This is a halogen C-LP unit. UPDATE I've been reading this https://www.nist.gov/sites/default/files/documents/ncnr/Microscope_Nikon_Eclipse_E600POL-0000.pdf and it seems the right most filter makes the light much more white. Second to rear filter appears to reduce glare?

    Parfocal? I've never used one of these before so I don't know if this is normal. If I focus the 4x Apo objective, then switch to the 10x Apo objective, I need to use the coarse focus dial to bring the subject into focus. I'm a little surprised by that. I thought these things were supposed to be parfocal. Switching from 10x to 20x Apo objective requires just a slight adjustment of the fine focus dial. And MAN is that objective close to the subject. WD 1.0 is narrow! Switching from 20x to 40x Apo is even closer with WD .14. I think I'll mostly hang out at 20x until I'm more comfortable and absolutely sure I won't grab the coarse focus knob by accident. Dang.

    Coarse focus lock. I'm pretty sure my coarse focus lock is broken, but I may just not be using it correctly. I turn the clamp knob on the right away from the arrow (toward me) then focus, then turn it the other way (away from me). If I lower the stage and raise it with the 4x objective until it stops my subject is barely visible it is so far out of focus.


Which tests/procedures should I be following right now to ensure this scope is fully functional? Thanks.


Last edited by Jesse on Thu Aug 23, 2018 9:16 am; edited 2 times in total
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Pau
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Joined: 20 Jan 2010
Posts: 4463
Location: Valencia, Spain

PostPosted: Thu Aug 23, 2018 9:03 am    Post subject: Reply with quote

Quote:
Confocal?

Confocal is a completely different thing, the term you want is parfocal

Parfocality between objectives in a microscope usually is not perfect, requiring a bit of refocusing with the fine focus knob is normal, but if you do not see an image even defocused when changing to the next magnification you have a parfocality issue. If so, first test if the objectives are well screwed in the nosepiece and he eyepieces are well placed all its way down. Later set the dioptric adjustments of the eyepieces o zero to later adjust only the diopter difference between your eyes.
Because you are inexperienced to begin I recommend you two booklets available to download from Zeiss and other internet sites:
"Microscopy from the very beginning"
"The clean microscope"

And be aware that your biological microscope is designed for transparent samples mounted with a 0.17mm cover glass. Only the 4X and in some cases the 10X are adequate for uncovered specimens.

The orangeish light is normal with halogen lamps when set at low power, you can correct it to some extent with a blue filter (most microscopes include it). At high power it must be whiter but likely it could burn your eyes at bright field low magnification, to avoid it don't close the diaphragm, use ND filters.
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Jesse



Joined: 11 Dec 2016
Posts: 64

PostPosted: Thu Aug 23, 2018 9:24 am    Post subject: Reply with quote

Pau wrote:

Parfocality between objectives in a microscope usually is not perfect, requiring a bit of refocusing with the fine focus knob is normal, but if you do not see an image even defocused when changing to the next magnification you have a parfocality issue.


I count three and a half turns of the fine focus knob from 4x -> 10x. Normal?
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Jesse



Joined: 11 Dec 2016
Posts: 64

PostPosted: Thu Aug 23, 2018 9:28 am    Post subject: Reply with quote

Pau wrote:

And be aware that your biological microscope is designed for transparent samples mounted with a 0.17mm cover glass. Only the 4X and in some cases the 10X are adequate for uncovered specimens.


Ok, but why? Is that a risk of damage thing or a refractive index thing or what?
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Pau
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Joined: 20 Jan 2010
Posts: 4463
Location: Valencia, Spain

PostPosted: Thu Aug 23, 2018 11:48 am    Post subject: Reply with quote

Jesse wrote:
I count three and a half turns of the fine focus knob from 4x -> 10x. Normal?

To me it seems clearly excessive, half turn or even full turn for low magnification could be OK
Jesse wrote:
Pau wrote:

And be aware that your biological microscope is designed for transparent samples mounted with a 0.17mm cover glass. Only the 4X and in some cases the 10X are adequate for uncovered specimens.


Ok, but why? Is that a risk of damage thing or a refractive index thing or what?

Aside the risk of damage due to the small WD there is an optical factor: your objectives have written on them /0.17 because the cover is part of the optical system, as you suggest due to the different RI of glass and air. Without cover you'll get spherical aberration degrading the image, it scales exponentially with the objective NA. It becomes noticeable from 0.3 upwards and it's so critical at 0.95 that your high magnification objectives have a correction collar to fine tune small variations in cover glass thickness.
And it's also important to have a flat and thin sample, beware that some objectives have WD smaller then the cover thickness.
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Jesse



Joined: 11 Dec 2016
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PostPosted: Thu Aug 23, 2018 11:57 am    Post subject: Reply with quote

Pau wrote:

Aside the risk of damage due to the small WD there is an optical factor: your objectives have written on them /0.17 because the cover is part of the optical system, as you suggest due to the different RI of glass and air. Without cover you'll get spherical aberration degrading the image, it scales exponentially with the objective NA. It becomes noticeable from 0.3 upwards and it's so critical at 0.95 that your high magnification objectives have a correction collar to fine tune small variations in cover glass thickness.
And it's also important to have a flat and thin sample, beware that some objectives have WD smaller then the cover thickness.


I may be seeing this effect. I have some plastic toy microscope slides that are pre-prepared. The 4x, 10x, and 20x objectives seem to work fine with them, but the 40x and 60x objectives look "hazy". I'll have to try with some glass slides and coverslips. I bought some, I just haven't prepared anything for them yet.
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Jesse



Joined: 11 Dec 2016
Posts: 64

PostPosted: Thu Aug 23, 2018 11:58 am    Post subject: Reply with quote

Pau wrote:

Because you are inexperienced to begin I recommend you two booklets available to download from Zeiss and other internet sites:
"Microscopy from the very beginning"


Thanks for this. Very helpful. I now feel like I have a fairly decent grasp on Kohler illumination. Image quality has improved quite a bit.
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Jesse



Joined: 11 Dec 2016
Posts: 64

PostPosted: Thu Aug 23, 2018 12:05 pm    Post subject: Reply with quote

One weird thing I've noticed is that my stage drifts to the right sometimes. I won't be touching anything and I'll notice the image slowly scrolling to the right. It does this for a few seconds and then stops. What could be causing this?

I've also noticed a bit of play when I touch the stage movement rod. If I push forward and backward gently I can wiggle the stage.
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