Correct Allignment of a Series 4 Stand

Here you can discuss different microscopic techniques and illumination methods, such as Brightfield, Darkfield, Phase Contrast, DIC, Oblique illumination, etc.
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apochronaut
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Joined: Fri May 15, 2015 12:15 am

Re: Correct Allignment of a Series 4 Stand

#31 Post by apochronaut » Tue Aug 04, 2020 12:34 pm

Is that accurate colour? The background should be grey. The other out of kilter aspect of the image are the chroma in the background. Your condenser is not focused properly. Keeping the objective in focus on the epithelium cells, close the field diaphragm, center the image of the closed aperture, then focus the condenser so the aperture is as sharp as possible. The closed aperture has a diffracted border, so there is a lot chroma. When focusing, there should be a point where the colour of the edge of the field diaphragm will change from red into purple. That is about best focus. Open the aperture all the way then. Make sure the aperture opens beyond the edges of the visual field or you have a partially frozen diaphragm. With the condenser properly focused and the field aperture wide open younushould then have a grey background and very even in tone with a 20.

You are getting an undue amount of ca around the cells, I think because the sample and cover glass may be too thick. Thin the sample out as much as possible and make sure your cover glass is .17.

Try all that and post another picture.

apochronaut
Posts: 3637
Joined: Fri May 15, 2015 12:15 am

Re: Correct Allignment of a Series 4 Stand

#32 Post by apochronaut » Thu Aug 06, 2020 12:04 pm

I had a thought about this and I may have asked you before, so apologies for the repeat but:. does your microscope have the compensating optic in the throat above the nosepiece?

Timemaster1212
Posts: 121
Joined: Sun Dec 03, 2017 12:21 am

Re: Correct Allignment of a Series 4 Stand

#33 Post by Timemaster1212 » Thu Aug 06, 2020 5:47 pm

Sorry for the late replies! Yes i do have the intermediate lens on my stand, and you were right i was out of focus apo. I found a hydra while checking on one of my pond samples, and while not the best test (nearly all of the halos are debris out of focus), to my untrained eye it appears to have a much more even background. Later today I will attempt the test again with cheeck cells.

Image

Apo, do you happen to have a reference image so i know exactly what effect i am looking for?

apochronaut
Posts: 3637
Joined: Fri May 15, 2015 12:15 am

Re: Correct Allignment of a Series 4 Stand

#34 Post by apochronaut » Thu Aug 06, 2020 7:05 pm

I will look. i still have the problem with my laptop frozen and all my picture files and the upload program on it. I haven't had time to sort that out.

apochronaut
Posts: 3637
Joined: Fri May 15, 2015 12:15 am

Re: Correct Allignment of a Series 4 Stand

#35 Post by apochronaut » Thu Aug 06, 2020 10:53 pm

Your images are improving greatly and this last with a relatively thick dpecimen too! Still the brownish background. You didn't mention if it is faithfull to what you are seeing in the eyepieces? Knowing what the objectives are helps too. I know they are dark phase but what is the contrast? Are they Dark M?

Timemaster1212
Posts: 121
Joined: Sun Dec 03, 2017 12:21 am

Re: Correct Allignment of a Series 4 Stand

#36 Post by Timemaster1212 » Fri Aug 07, 2020 5:31 pm

So, the objectives on my stand are Dark M Phase contrast. Also, the view through the eyepiece is more grey than the image, which makes me wonder if the browning is a white balance issue, my canon is set to automatic. I am making another swab slide to show more properties of the image without being to thick. I will also set the WB to tungsten, see what happens then.

Timemaster1212
Posts: 121
Joined: Sun Dec 03, 2017 12:21 am

Re: Correct Allignment of a Series 4 Stand

#37 Post by Timemaster1212 » Fri Aug 07, 2020 7:40 pm

Image

Alrighty, i think i figured it out haha, camera issue LOL. I think this is the best image yet, let me know what yall think.

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