Trying out my new objective

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LouiseScot
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Trying out my new objective

#1 Post by LouiseScot » Sun Apr 11, 2021 4:00 pm

It's a Nikon CF Fluorite F100-UV 100/1.30 glycerine immersion. I tried it with some pollen mounted in glycerine and with glycerine immersion. It seemed to work ok but the images weren't very interesting. I also tried it with glycerine immersion on some diatoms mounted in Pleurax. Not homogenous, but it seemed to still work ok. I guess not optimal, but still.

So some diatoms :)

Cropped and resized to 77%
F100 Med Diatom _Crop_77pc_00001.jpg
F100 Med Diatom _Crop_77pc_00001.jpg (86.17 KiB) Viewed 5432 times


Cropped and resized to 50%
F100 Med Diatom _Crop_50pc_00010.jpg
F100 Med Diatom _Crop_50pc_00010.jpg (95.06 KiB) Viewed 5432 times


Cropped and resized to 61%
F100 Med Diatom _Crop61pc_00021.jpg
F100 Med Diatom _Crop61pc_00021.jpg (63.94 KiB) Viewed 5432 times


Cropped and resized to 41%
F100 Med Diatom _Crop41pc_00027.jpg
F100 Med Diatom _Crop41pc_00027.jpg (113.67 KiB) Viewed 5432 times

All taken with my Gpcam3 178M at 415nm on my Swift380T

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

Hobbyst46
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Re: Trying out my new objective

#2 Post by Hobbyst46 » Sun Apr 11, 2021 4:25 pm

Very lovely high resolution images.
Once again demonstrate immersion power. ;)
Oblique, aren't they ? how did you get the oblique illumination ?
Since that is a Nikon Cf, would be interesting to see color images as well.
Incidentally, is that a Fluorite objective ? the CF and F do not mean fluorite in this case, I think.

LouiseScot
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Re: Trying out my new objective

#3 Post by LouiseScot » Sun Apr 11, 2021 4:34 pm

Hobbyst46 wrote:
Sun Apr 11, 2021 4:25 pm
Very lovely high resolution images.
Once again demonstrate immersion power. ;)
Oblique, aren't they ? how did you get the oblique illumination ?
Since that is a Nikon Cf, would be interesting to see color images as well.
Thanks! Yeah, I was a bit worried that the glycerine immersion might mess up the Pleurax mount images completely but I think they are acceptable. I've ordered some borosilicate 3.3 coverslips and will see if they make things better - or worse! (with my own slides).
I'm using an LED mounted on a heatsink under the condenser so I can move it it around to get oblique illumination.
I'll try it with the dslr. Also the 40/0.85 Fluor I recently bought. I've been carried away with the mono imaging!

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

LouiseScot
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Re: Trying out my new objective

#4 Post by LouiseScot » Sun Apr 11, 2021 5:52 pm

Comparing the Fluorite Petroneis with one taken by a 100x oil achromat. The image is much reduced and I rotated the achromat one to be about the same orientation as the Nikon Fluorite:

Petroneis_100xAchro_vsFluorCrop50pc_Red0017.jpg
Petroneis_100xAchro_vsFluorCrop50pc_Red0017.jpg (102.71 KiB) Viewed 5411 times

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

Hobbyst46
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Re: Trying out my new objective

#5 Post by Hobbyst46 » Sun Apr 11, 2021 6:08 pm

Getting better all the time !

LouiseScot
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Re: Trying out my new objective

#6 Post by LouiseScot » Sun Apr 11, 2021 6:17 pm

Hobbyst46 wrote:
Sun Apr 11, 2021 6:08 pm
Getting better all the time !
That was just a quick comparison using the previous Petroneis image I took. You'd expect the Nikon fluorite to be better than the Swift achromat! Still, I think the Nikon 100x was a bargain! Got it from the same Japanese seller as the Nikon 40/0.85 fluor. Both good, and both good value for money :) The 100x was actually a lot cheaper than the 40x!

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

MichaelG.
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Re: Trying out my new objective

#7 Post by MichaelG. » Sun Apr 11, 2021 6:26 pm

LouiseScot wrote:
Sun Apr 11, 2021 5:52 pm
Petroneis_100xAchro_vsFluorCrop50pc_Red0017.jpg
.
Louise
Lovely image from that Fluorite
... but what's with the 'staircase' effect [very evident at upper left] ?

MichaelG.
Too many 'projects'

DonSchaeffer
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Re: Trying out my new objective

#8 Post by DonSchaeffer » Sun Apr 11, 2021 6:30 pm

Very impressive. Do you find that colors seem to wash out at high magnification?

LouiseScot
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Re: Trying out my new objective

#9 Post by LouiseScot » Sun Apr 11, 2021 6:35 pm

DonSchaeffer wrote:
Sun Apr 11, 2021 6:30 pm
Very impressive. Do you find that colors seem to wash out at high magnification?
They are monochrome images taken with a monochrome astro camera with illumination from a 415nm (blue-violet) , 3W LED :)
I will take some colour ones with the dslr next time.

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

LouiseScot
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Re: Trying out my new objective

#10 Post by LouiseScot » Sun Apr 11, 2021 6:37 pm

MichaelG. wrote:
Sun Apr 11, 2021 6:26 pm
LouiseScot wrote:
Sun Apr 11, 2021 5:52 pm
Petroneis_100xAchro_vsFluorCrop50pc_Red0017.jpg
.
Louise
Lovely image from that Fluorite
... but what's with the 'staircase' effect [very evident at upper left] ?

MichaelG.
Thanks. Not sure what you mean re 'staircase??

ps Oh think I see - not sure. May be the oblique illumination else, possibly, refraction effects because of the mismatch between the glycerine, glass and mounting medium. I only got the objective yesterday and will have to try it out in different situations. I didn't notice the same anomaly in the other images above.

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

LouiseScot
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Re: Trying out my new objective

#11 Post by LouiseScot » Sun Apr 11, 2021 7:05 pm

LouiseScot wrote:
Sun Apr 11, 2021 6:37 pm
MichaelG. wrote:
Sun Apr 11, 2021 6:26 pm
LouiseScot wrote:
Sun Apr 11, 2021 5:52 pm
Petroneis_100xAchro_vsFluorCrop50pc_Red0017.jpg
.
Louise
Lovely image from that Fluorite
... but what's with the 'staircase' effect [very evident at upper left] ?

MichaelG.
Thanks. Not sure what you mean re 'staircase??

ps Oh think I see - not sure. May be the oblique illumination else, possibly, refraction effects because of the mismatch between the glycerine, glass and mounting medium. I only got the objective yesterday and will have to try it out in different situations. I didn't notice the same anomaly in the other images above.

Louise
This is a different individual I imaged from the same slide at more or less the same time:
F100 Med Diatom _Crop70pc.jpg
F100 Med Diatom _Crop70pc.jpg (79.22 KiB) Viewed 5393 times
It shows the same effect. It looks like it's to do with that species- maybe they are quite deep?

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

LouiseScot
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Re: Trying out my new objective

#12 Post by LouiseScot » Sun Apr 11, 2021 8:30 pm

Before I took stills (selection above) I made a short .avi of a diatom on another slide. I recorded for 2 1/2 mins but it only plays for 18 secs. Must be a setting... I should have changed the size/dimensions too so it was the full 3040x2048... Next time I'll try harder to get it right :D It's unedited.

https://youtu.be/eiEKgfmMj_4


Of course you can pause it and inspect at any point :)


Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

MichaelG.
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Re: Trying out my new objective

#13 Post by MichaelG. » Mon Apr 12, 2021 8:46 am

LouiseScot wrote:
Sun Apr 11, 2021 7:05 pm
It shows the same effect. It looks like it's to do with that species- maybe they are quite deep?
.

I’ve just found this:
https://www.researchgate.net/publicatio ... _do_Brasil

Have a look at Figure 3 ... it does seem to show a similar effect

Interesting !!

MichaelG.
Too many 'projects'

LouiseScot
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Re: Trying out my new objective

#14 Post by LouiseScot » Mon Apr 12, 2021 9:05 am

MichaelG. wrote:
Mon Apr 12, 2021 8:46 am
LouiseScot wrote:
Sun Apr 11, 2021 7:05 pm
It shows the same effect. It looks like it's to do with that species- maybe they are quite deep?
.

I’ve just found this:
https://www.researchgate.net/publicatio ... _do_Brasil

Have a look at Figure 3 ... it does seem to show a similar effect

Interesting !!

MichaelG.
I'm sure there's a simple explanation...

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

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Rossf
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Re: Trying out my new objective

#15 Post by Rossf » Mon Apr 12, 2021 9:22 am

Hey LouiseScot your doing a great job of upgrading the Swift-your images just keep getting better and you seem to really know what you want in an objective-thats why I stuck with finite systems-I like to “shop” for optics from a variety of makers-I got a few good objectives from Japanese sellers on eBay recently as well-your enthusiasm is infectious.
Regards ross

LouiseScot
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Re: Trying out my new objective

#16 Post by LouiseScot » Mon Apr 12, 2021 9:29 am

Rossf wrote:
Mon Apr 12, 2021 9:22 am
Hey LouiseScot your doing a great job of upgrading the Swift-your images just keep getting better and you seem to really know what you want in an objective-thats why I stuck with finite systems-I like to “shop” for optics from a variety of makers-I got a few good objectives from Japanese sellers on eBay recently as well-your enthusiasm is infectious.
Regards ross
Thanks, Ross! Well I just went with the Nikon cf range which don't need any extra correction and, of course, they are an excellent make, as well as being parfocal. Coming across the two fluors recently was good luck, I think! As I mentioned, the fluorite glycerine immersion lens was a real bargain. I also have a diy infinity system based on a macro focus rail but I've rather neglected it lately. I will do something useful with it soon! So much to do, so little time!

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

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Rossf
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Re: Trying out my new objective

#17 Post by Rossf » Mon Apr 12, 2021 9:49 am

I think most of us here a guilty of neglecting a piece of gear for a while..especially while enthusiastically going down a particular rabbit hole! But it’s always good to reconnect with that different scope down the road-I think that helps keep enthusiasm going sometimes..just taking a side step to another optical adventure-I’ve got a sneaking suspicion your fascination with the optical effects of glass goes back to childhood-mine did! I was always looking through glass-I grew up in the 70’s so had lovely chunky ashtrays etc to observe how light behaves as it goes through-playing with family movie cameras and filters(!)-they seemed magic at 8 years old! Well still do...

LouiseScot
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Re: Trying out my new objective

#18 Post by LouiseScot » Mon Apr 12, 2021 10:12 am

Rossf wrote:
Mon Apr 12, 2021 9:49 am
I think most of us here a guilty of neglecting a piece of gear for a while..especially while enthusiastically going down a particular rabbit hole! But it’s always good to reconnect with that different scope down the road-I think that helps keep enthusiasm going sometimes..just taking a side step to another optical adventure-I’ve got a sneaking suspicion your fascination with the optical effects of glass goes back to childhood-mine did! I was always looking through glass-I grew up in the 70’s so had lovely chunky ashtrays etc to observe how light behaves as it goes through-playing with family movie cameras and filters(!)-they seemed magic at 8 years old! Well still do...
I'm a child of the 50's :) My first microscope arrived when I persuaded my mum to get one on special offer on a cereal packet. Can't remember which one - think it was Shredded Wheat. It was a plasticky thing but quite good, nevertheless. It was the first time I was able to see protozoa and such :) Then when I was 18 I left school and went to work in a big pharma quality control / bacteriology lab. I used to collect various environmental samples and make agar plates, often with added horse blood. Make slides, Gram stain them and examine them under a microscope. I loved it! But moved into biochemistry a bit later. More recently (2009-13) I was doing research into a Rett Syndrome (a genetic disease) and was doing patch clamping of mouse brain slices using water immersion/IR-DIC, and further examination of antibody staining with a laser scanning confocal microscope. Got my name on a paper too :) I'm retired now. I have several refractor telescopes also but the weather and sky conditions here are not really conducive, so I've mostly moved back to microscopes - back to where I started :) Oddly enough, through all the time I've been using microscopes it's only since I've been a member here that I've started delving into the deeper aspects of microscopes and objectives, and lighting etc. etc. There's always more to learn!

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

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Rossf
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Re: Trying out my new objective

#19 Post by Rossf » Mon Apr 12, 2021 11:21 am

👍

Hobbyst46
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Re: Trying out my new objective

#20 Post by Hobbyst46 » Mon Apr 12, 2021 1:32 pm

LouiseScot wrote:
Mon Apr 12, 2021 9:05 am
MichaelG. wrote:
Mon Apr 12, 2021 8:46 am
LouiseScot wrote:
Sun Apr 11, 2021 7:05 pm
It shows the same effect. It looks like it's to do with that species- maybe they are quite deep?
.

I’ve just found this:
https://www.researchgate.net/publicatio ... _do_Brasil

Have a look at Figure 3 ... it does seem to show a similar effect

Interesting !!

MichaelG.
I'm sure there's a simple explanation...

Louise
The thickness of the diatom is being measured right now...

LouiseScot
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Location: Scotland

Re: Trying out my new objective

#21 Post by LouiseScot » Mon Apr 12, 2021 1:46 pm

Hobbyst46 wrote:
Mon Apr 12, 2021 1:32 pm
LouiseScot wrote:
Mon Apr 12, 2021 9:05 am
MichaelG. wrote:
Mon Apr 12, 2021 8:46 am


.

I’ve just found this:
https://www.researchgate.net/publicatio ... _do_Brasil

Have a look at Figure 3 ... it does seem to show a similar effect

Interesting !!

MichaelG.
I'm sure there's a simple explanation...

Louise
The thickness of the diatom is being measured right now...
Says 30um here http://nordicmicroalgae.org/taxon/Petroneis%20humerosa
:)
Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

Hobbyst46
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Re: Trying out my new objective

#22 Post by Hobbyst46 » Mon Apr 12, 2021 1:52 pm

Thickness - neither width nor height.

LouiseScot
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Re: Trying out my new objective

#23 Post by LouiseScot » Mon Apr 12, 2021 1:59 pm

Hobbyst46 wrote:
Mon Apr 12, 2021 1:52 pm
Thickness - neither width nor height.
? thickness = height :)

The source gives length, width and height - good enough?
A 'thickness' of 30um seems quite a lot when width is also 30um

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

Hobbyst46
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Re: Trying out my new objective

#24 Post by Hobbyst46 » Mon Apr 12, 2021 2:40 pm

LouiseScot wrote:The source gives length, width and height - good enough?A 'thickness' of 30um seems quite a lot when width is also 30um
I stand corrected, sorry !

So here is the measurement. Recorded with the 16x0.32 Plan objective.
Photo 1 shows the LED strip stand. Laid on the microscope stage (usually the stereo microscope, but for the present purpose - on the compound). The slide holder arm is taped to the stage, to keep it away. The LED strip is vertical, and the blackened slide is laid on the stage beneath the LEDs.
The blackened slide is, in fact, a strew slide of gently cleaned diatoms (photo 2). From this Smaug's treasure heap I peak the gem, and place it in a quiet vacant dark area (photo 3). I lies vertically, almost, slanted just a bit, the apex points up (photo 4). The stage micrometer (photo 5) imaged at the same conditions provide the measurement scale.

The length of the diatom is estimated on the basis of its siblings (one of them is visible in photo 2) to be 60-65um.
The width is not reliable, since the diatom is slanted.

The thickness, according to photos 3 and 4, is estimated to be ~15 um. Could be more, though. I have to improve the illumination for better accuracy.

The gentle cleaning can preserve the complete frustule.
So these are complete frustules. This conclusion is somewhat supported by the fact, that when an inappropriate mountant was used - Norland 61 adhesive rather than Pleurax - large air bubbles stayed within this very frustule.
Attachments
1) LED stand for handling dry diatoms on a blackened slide.jpg
1) LED stand for handling dry diatoms on a blackened slide.jpg (105.78 KiB) Viewed 5270 times
2) Diatom keeper black slide 2.JPG
2) Diatom keeper black slide 2.JPG (169.38 KiB) Viewed 5270 times
3) Resized diatom.jpg
3) Resized diatom.jpg (47.13 KiB) Viewed 5270 times
4) Cropped diatom.jpg
4) Cropped diatom.jpg (55.92 KiB) Viewed 5270 times
5) Stage micrometer view under 16X objective. Line spacing = 10 um.JPG
5) Stage micrometer view under 16X objective. Line spacing = 10 um.JPG (66.87 KiB) Viewed 5270 times

LouiseScot
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Re: Trying out my new objective

#25 Post by LouiseScot » Mon Apr 12, 2021 3:36 pm

Hobbyst46 wrote:
Mon Apr 12, 2021 2:40 pm
LouiseScot wrote:The source gives length, width and height - good enough?A 'thickness' of 30um seems quite a lot when width is also 30um
I stand corrected, sorry !

So here is the measurement. Recorded with the 16x0.32 Plan objective.
Photo 1 shows the LED strip stand. Laid on the microscope stage (usually the stereo microscope, but for the present purpose - on the compound). The slide holder arm is taped to the stage, to keep it away. The LED strip is vertical, and the blackened slide is laid on the stage beneath the LEDs.
The blackened slide is, in fact, a strew slide of gently cleaned diatoms (photo 2). From this Smaug's treasure heap I peak the gem, and place it in a quiet vacant dark area (photo 3). I lies vertically, almost, slanted just a bit, the apex points up (photo 4). The stage micrometer (photo 5) imaged at the same conditions provide the measurement scale.

The length of the diatom is estimated on the basis of its siblings (one of them is visible in photo 2) to be 60-65um.
The width is not reliable, since the diatom is slanted.

The thickness, according to photos 3 and 4, is estimated to be ~15 um. Could be more, though. I have to improve the illumination for better accuracy.

The gentle cleaning can preserve the complete frustule.
So these are complete frustules. This conclusion is somewhat supported by the fact, that when an inappropriate mountant was used - Norland 61 adhesive rather than Pleurax - large air bubbles stayed within this very frustule.
Wow! So many you have their Bilbo! So is that actually a Petroneis sideways on? I'd say it was more like 20um just measuring with a ruler on screen. So 20x60 isn't far removed from 30x70. Anyway, I'm still not sure of the causes of the image aberrations above...

Louise
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

Hobbyst46
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Re: Trying out my new objective

#26 Post by Hobbyst46 » Mon Apr 12, 2021 3:43 pm

LouiseScot wrote:
Mon Apr 12, 2021 3:36 pm
Wow! So many you have their Bilbo! So is that actually a Petroneis sideways on? I'd say it was more like 20um just measuring with a ruler on screen. So 20x60 isn't far removed from 30x70. Anyway, I'm still not sure of the causes of the image aberrations above...
Yes, it is sideways on. I flipped it over with the tip of the glass fiber then back on its top. The exact thickness is really an estimate. Since it was not exactly perpendicular to the plane of the slide.

LouiseScot
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Re: Trying out my new objective

#27 Post by LouiseScot » Mon Apr 12, 2021 3:47 pm

Hobbyst46 wrote:
Mon Apr 12, 2021 3:43 pm
LouiseScot wrote:
Mon Apr 12, 2021 3:36 pm
Wow! So many you have their Bilbo! So is that actually a Petroneis sideways on? I'd say it was more like 20um just measuring with a ruler on screen. So 20x60 isn't far removed from 30x70. Anyway, I'm still not sure of the causes of the image aberrations above...
Yes, it is sideways on. I flipped it over with the tip of the glass fiber then back on its top. The exact thickness is really an estimate. Since it was not exactly perpendicular to the plane of the slide.
I couldn't do that! My hands would probably shake too much!
A Nikon CF plan 20x; A Swift 380T; A DIY infinity corrected focus rail system with a 40x/0.65 Olympus Plan, a 10x/0.30 Amscope Plan Fluor, and a 20x/0.75 Nikon Plan Apo

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