Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

Everything relating to microscopy hardware: Objectives, eyepieces, lamps and more.
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FungusMan
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#31 Post by FungusMan » Wed Sep 08, 2021 3:49 am

MichaelG. wrote:
Tue Sep 07, 2021 6:15 am
I have been browsing through that first-nature website, in search of a good spore to use as a test specimen:

This looks promising : https://www.first-nature.com/fungi/aleuria-aurantia.php

MichaelG.

.
Edit: __ See also Fig.2 here: http://www.leifgoodwin.co.uk/General/Microscopy.html
Most of the images are about 300pixels wide. If I resize and post-edit quality get close to those... so I am still thinking maybe I have to reconsider that at x1000 I cant expect model images.

FungusMan
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#32 Post by FungusMan » Wed Sep 08, 2021 4:03 am

Many thanks to those who had helped with EFSC on the 750D, known as silent mode (frame with s icon at the corner). As you correctly pointed this function is available only when viewing with the viewfinder. Focusing my image with the viewfinder is quite impractible and cant be considered. I have to rely focusing on the screen. In this mode I can shoot with
I) timer
ii) touch screen
iii) snapshot chord (RS60)

Would any of the these three prevent the said vibrations?

viktor j nilsson
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#33 Post by viktor j nilsson » Wed Sep 08, 2021 4:21 am

FungusMan wrote:
Wed Sep 08, 2021 4:03 am
Many thanks to those who had helped with EFSC on the 750D, known as silent mode (frame with s icon at the corner). As you correctly pointed this function is available only when viewing with the viewfinder. Focusing my image with the viewfinder is quite impractible and cant be considered. I have to rely focusing on the screen. In this mode I can shoot with
I) timer
ii) touch screen
iii) snapshot chord (RS60)

Would any of the these three prevent the said vibrations?
Who said it's only available when using the viewfinder? It's the opposite: silent mode is only available when using live view and looking at the LCD screen.

FungusMan
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#34 Post by FungusMan » Wed Sep 08, 2021 4:42 am

Here are some spores of a Gasteromycete fungus (likely Mycenastrum / Scleroderma) approx the best I can get from the EOS750D plus the adapter fitted in the eyepiece. Resized, sharpened and some mild contrast adjustment. The spores are about 18um - as we discussed, these spherical bodies cant be focused throughout, but there detail of the spines (c. 1.5um) that are curved and sharp-pointed.

So maybe I should first buy a trinocular and see if we get any better

This is my favourite
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FungusMan
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#35 Post by FungusMan » Wed Sep 08, 2021 4:45 am

If resized to 300px
IMG_3570ss.jpg
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MichaelG.
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#36 Post by MichaelG. » Wed Sep 08, 2021 6:32 am

FungusMan wrote:
Wed Sep 08, 2021 3:49 am
MichaelG. wrote:
Tue Sep 07, 2021 6:15 am
… in search of a good spore to use as a test specimen:

This looks promising : … aleuria-aurantia
Most of the images are about 300pixels wide. If I resize and post-edit quality get close to those... so I am still thinking maybe I have to reconsider that at x1000 I cant expect model images.
Sorry if what I wrote wasn’t sufficiently clear:
I was not trying to imply that the referenced pictures should set a stretch-target for image quality
… but simply that Aleuria aurantia looks like a suitable test specimen.

MichaelG.
Too many 'projects'

Alexander
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#37 Post by Alexander » Wed Sep 08, 2021 6:40 am

The problem may be your eye-piece adapter. They are quite afocal. The issue increases with the power of the objective. A 100x objective may simply be out of focus. Try a dry and well fixed sample without cover glas. This allows the objective to be closer to the sample.


About LiveView: LiveView always uses EFCS. The mechanical shutter needs to be open to produce a picture in LiveView. For viewfinder use EFCS can be activated as well.

FungusMan
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#38 Post by FungusMan » Wed Sep 08, 2021 11:10 am

Alexander wrote:
Wed Sep 08, 2021 6:40 am
The problem may be your eye-piece adapter. They are quite afocal. The issue increases with the power of the objective. A 100x objective may simply be out of focus. Try a dry and well fixed sample without cover glas. This allows the objective to be closer to the sample.


About LiveView: LiveView always uses EFCS. The mechanical shutter needs to be open to produce a picture in LiveView. For viewfinder use EFCS can be activated as well.
Yes I am not happy about it. It does a mediocre job, which well, it is better than nothing but not close to perfect. I had no other cheap option in my binocular microscope. Now I sourced a trinocular for 1000USD including transport and I might go for it. I also think my objectives are doing fine. I cleaned thoroughly the x63 and its performing satisfactorily. The problem is imaging.

FungusMan
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#39 Post by FungusMan » Wed Sep 08, 2021 11:11 am

Alexander wrote:
Wed Sep 08, 2021 6:40 am
About LiveView: LiveView always uses EFCS. The mechanical shutter needs to be open to produce a picture in LiveView. For viewfinder use EFCS can be activated as well.
Good to know - thanks

Gatorengineer64
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#40 Post by Gatorengineer64 » Wed Sep 08, 2021 5:10 pm

I recieved by Nikon Achro yesterday for my eclipse 600. I am 99 percent certain that your issues are chromatic aberation (notice how colorful the out of focus areas are). I do not have an APO 100 I have a plan Achro nikon100x and the Achro condensor really made a difference. If you buy one on ebay and it doesnt work worst is that you are out $20 plus shipping if you resell it. Its a cheap experiment and I am confident it will fix your issue. The axioscope and zeiss optics are excellent.

MicroBob
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#41 Post by MicroBob » Wed Sep 08, 2021 5:57 pm

Alexander wrote:
Wed Sep 08, 2021 6:40 am
The problem may be your eye-piece adapter. They are quite afocal. The issue increases with the power of the objective. A 100x objective may simply be out of focus. Try a dry and well fixed sample without cover glas. This allows the objective to be closer to the sample.
Good point! The objective has to be used in eactly the focus position as when observing. If the camera adapter is wrong, one can get a sharp image at a different focus setting, but the objective is not used as it was calculated.
Alexander wrote:
Wed Sep 08, 2021 6:40 am
About LiveView: LiveView always uses EFCS. The mechanical shutter needs to be open to produce a picture in LiveView. For viewfinder use EFCS can be activated as well.
Depends on the camera and setting: Many cameras (even DSLMs without mirror) close and reopen the shuter before exposure. My old Sony NEX 5 gave quite a whack due to this, blurring the pictures. The NEX 5N had EFSC and got rid of the problem, a nice old camera to use even today.

Bob

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patta
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#42 Post by patta » Thu Sep 09, 2021 7:58 am

Agree with all comments above... but two more issues, maybe I'm wrong...

1) sure the last spores with spines are 18 micron diameter? I would guess 9!
There is 100x objective and 2x Amscope adapter, total 200x. The pixels of 750D are 3,7 micron; thus one pixel is 3,7/200 = 0,0185 micron.
Max resolution for 100x 1.30 is about 0.2 micron, and will be about 11 pixels on the sensor. So, maybe optics is ok, just too much expectations and pixel peeping.

2) is.. the condenser... fully open or not? At NA 1.30, depth of field is about 0.2 micron - less than a spine in the spore. With fully open condenser at NA 1,30 (or even just dry 0.90), only a thin rim would be in focus, the rest totally blurred? From the images, depth of focus looks much deeper, hence guess that the condenser is not at MAX, thus resolution is reduced.
IMG_9467ss.jpg
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MichaelG.
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#43 Post by MichaelG. » Thu Sep 09, 2021 8:39 am

patta wrote:
Thu Sep 09, 2021 7:58 am

2) is.. the condenser... fully open or not? At NA 1.30, depth of field is about 0.2 micron - less than a spine in the spore. With fully open condenser at NA 1,30 (or even just dry 0.90), only a thin rim would be in focus, […]
I agree … I think the illumination settings are generating artefacts

MichaelG.
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#44 Post by patta » Mon Sep 13, 2021 11:30 am

I've just had too a look at spores from some puff mushroom (Gasteromycete?), similar as from previous posts.
The spines are indeed pretty hard to see well.

With water mount (spores sprinkled on a droplet of water, mix, cover with coverslip) the result is so-so for the 100x oil immersion.
A net improvement was to use oil as mountant instead of water (a drop of oil on the slide; sprinkle with spores, mix, cover with coverslip), Much clearer image than the water. The spores mixed well in oil.

Sure a proper mounting medium like Canada balsam would be better and more durable; but oil is easy and quick.

FungusMan
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#45 Post by FungusMan » Thu Sep 16, 2021 6:59 pm

It never crossed my mind to mount in oil! I try that! Thanks for yr feedback

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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#46 Post by FungusMan » Thu Sep 16, 2021 7:00 pm

UPDATE: I bought a trinocular (Zeiss original second hand) and I will update you if there are improvements
Cheers

FungusMan
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Re: Images from x1000 and AxioLab Re - is image quality as expected or crap and need to upgrade?

#47 Post by FungusMan » Thu Sep 16, 2021 7:07 pm

patta wrote:
Thu Sep 09, 2021 7:58 am
Agree with all comments above... but two more issues, maybe I'm wrong...

1) sure the last spores with spines are 18 micron diameter? I would guess 9!
There is 100x objective and 2x Amscope adapter, total 200x. The pixels of 750D are 3,7 micron; thus one pixel is 3,7/200 = 0,0185 micron.
Max resolution for 100x 1.30 is about 0.2 micron, and will be about 11 pixels on the sensor. So, maybe optics is ok, just too much expectations and pixel peeping.

2) is.. the condenser... fully open or not? At NA 1.30, depth of field is about 0.2 micron - less than a spine in the spore. With fully open condenser at NA 1,30 (or even just dry 0.90), only a thin rim would be in focus, the rest totally blurred? From the images, depth of focus looks much deeper, hence guess that the condenser is not at MAX, thus resolution is reduced.
IMG_9467ss.jpg

1) I measured accurately with pixmetre and they are 12-15um (+/- 1) - Your technicaly hypothesis confirms with what we have been saying 'vaguely' that at x 100 (oil) the image quality should not be expected to be particularly sharp and detailed. So I must be moderately please with what I get so far. Now I bought a trinocular and see what improvement the quality of the images will get.

2) No, the Condenser aperture was about half or two thirds up. But Is Kohler illumination adjustment useful at x100 oil? It seems Ok for x400 and lower but no particular noticeable difference at x63 and x 100

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