Trying out a mono camera

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LouiseScot
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Trying out a mono camera

#1 Post by LouiseScot » Wed Mar 31, 2021 11:20 pm

I made an adapter that makes it simple to connect a small mono astro camera to an ocular port on the Swift 380T. That way I can leave the dslr/trinocular setup alone. It's 6MP and uses a Sony 178M sensor. The sensor itself is very small - 7.37 x 4.92mm, as are the pixels - 2.4um. Focus is critical and it's hard to move the slide as even the smallest turn of the xy adjuster can send the target whooshing off!
The adapter allows easy attachment/removal. I may have to fiddle a bit to get it exactly parfocal with the ocular in the other port but it's very close as is. Being mono, and with an AR glass window, it's sensitive to a wide spectral range: 350nm - 1050nm so there is the potential option to image in near UV (I'm not sure how the optics between the objective and the camera will behave in UV) as well as IR. However, I really want to try using it with a monochromatic visible light source to maximise resolution. I've ordered some blue and violet LEDs and they should give it a run for its money!
Meanwhile, here's a couple I took with the Swift white LED and slightly oblique light, using the Nikon 40x/0.85 fluor. Slightly different exposures and contrast enhancements - which is the better image? Answers on a postcard, please :)

Med_Nikon_40x_178M_Crop_00001.jpg
Med_Nikon_40x_178M_Crop_00001.jpg (45.1 KiB) Viewed 6829 times


Med_Nikon_40x_178M_Crop_00002.jpg
Med_Nikon_40x_178M_Crop_00002.jpg (41.31 KiB) Viewed 6829 times

I'll have to sort out a scale for it.

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

Javier
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Re: Trying out a mono camera

#2 Post by Javier » Thu Apr 01, 2021 12:37 pm

Nice!

Looking forward to seeing that diatom in LRGB. :lol:

LouiseScot
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Re: Trying out a mono camera

#3 Post by LouiseScot » Thu Apr 01, 2021 12:40 pm

Javier wrote:
Thu Apr 01, 2021 12:37 pm
Nice!

Looking forward to seeing that diatom in LRGB. :lol:
Thanks! And that's not going to happen! Anyway, it's white to start off with :)

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

MicroBob
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Re: Trying out a mono camera

#4 Post by MicroBob » Thu Apr 01, 2021 1:35 pm

Hi Louise,
for diatoms these monochrome cameras are very good. Microscope optics transmit down to about 360nm. Some mountants (e.g. Pleurax) don't transmit any light below 420nm or so. If you are more interested in UV use on diatoms you might google for the work of Peter Höbel, very useful.
Apart from a blue or near UV LED you could use an interference short pass filter.
What also helps a little is to use crossed polarizers. The image will become very dim when applying all these methods leading to a noisy image. To reduce the noise a couple of similar images can be calculated together, leveling out the noise.
Here a lot of high resolution diatom photography tricks are discussed: https://www.mikroskopie-forum.de/index. ... c=40111.45

I edited your image, levels and unsharp mask, how do you like it?

Bob
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Hobbyst46
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Re: Trying out a mono camera

#5 Post by Hobbyst46 » Thu Apr 01, 2021 2:01 pm

MicroBob wrote:
Thu Apr 01, 2021 1:35 pm
Here a lot of high resolution diatom photography tricks are discussed: https://www.mikroskopie-forum.de/index. ... c=40111.45
Bob, in that link I see a new (?) high RI resin named Speedax. Is it available of-the-shelf ?

LouiseScot
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Re: Trying out a mono camera

#6 Post by LouiseScot » Thu Apr 01, 2021 2:10 pm

Hobbyst46 wrote:
Thu Apr 01, 2021 2:01 pm
MicroBob wrote:
Thu Apr 01, 2021 1:35 pm
Here a lot of high resolution diatom photography tricks are discussed: https://www.mikroskopie-forum.de/index. ... c=40111.45
Bob, in that link I see a new (?) high RI resin named Speedax. Is it available of-the-shelf ?
It says 'available exclusively from me' - Georg Abele : i.e. mba Abele GmbH Nägeleshofstr. 80 73434 Aalen E-mail: thin-section@t-online.de
I didn't see a price....

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 a mono camera

#7 Post by LouiseScot » Thu Apr 01, 2021 8:35 pm

I did one this afternoon using the 486nm filter. The trouble with it is that it needs a very long exposure and there's a good chance of slight vibration ruining it. I think it was actually ok in the end. Using a 3W LED at a shorter wavelength will be a much better method, I'm sure. I measured up and added a scale bar. Then decided I should do it properly and used Fiji - so two scale bars for the price of one, as it were :) However, it returned 0.01mm rather than 10um - I'll have to look up how to change that. I suppose it's in the initial scale setup bit... I think I put the measurement in mm rather than um, um.

Med_Nikon_Fluor_40x_486nm_cropaScale100pc.jpg
Med_Nikon_Fluor_40x_486nm_cropaScale100pc.jpg (52.51 KiB) Viewed 6747 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 a mono camera

#8 Post by Hobbyst46 » Thu Apr 01, 2021 9:20 pm

The 486nm light has improved the image. Certainly a powerful LED will be better than filtration of white light. I cannot stand the blue light in my eyes though.
Is it easy to switch the LEDs on the Swift microscope, from white to blue? On my microscope it would need fabrication of a new LED illuminator.

LouiseScot
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Re: Trying out a mono camera

#9 Post by LouiseScot » Thu Apr 01, 2021 9:39 pm

Hobbyst46 wrote:
Thu Apr 01, 2021 9:20 pm
The 486nm light has improved the image. Certainly a powerful LED will be better than filtration of white light. I cannot stand the blue light in my eyes though.
Is it easy to switch the LEDs on the Swift microscope, from white to blue? On my microscope it would need fabrication of a new LED illuminator.
Thanks - I wasn't 100% sure about it. It's easier to leave the 1W Swift LED in place and just put another above it. I've already made separate 3W and 10W LEDs (mainly for DF). I just need to put the violet/blue LEDs on heatsinks and solder a short lead with a standard 5x2.1mm power socket. I've got power supplies set up which enable LED dimming.
By the way, did you look into the Speedax mountant?

ps I like your Pleurosigma in the other thread.

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 a mono camera

#10 Post by Hobbyst46 » Thu Apr 01, 2021 10:11 pm

LouiseScot wrote:
Thu Apr 01, 2021 9:39 pm
Hobbyst46 wrote:
Thu Apr 01, 2021 9:20 pm
The 486nm light has improved the image. Certainly a powerful LED will be better than filtration of white light. I cannot stand the blue light in my eyes though.
Is it easy to switch the LEDs on the Swift microscope, from white to blue? On my microscope it would need fabrication of a new LED illuminator.
Thanks - I wasn't 100% sure about it. It's easier to leave the 1W Swift LED in place and just put another above it. I've already made separate 3W and 10W LEDs (mainly for DF). I just need to put the violet/blue LEDs on heatsinks and solder a short lead with a standard 5x2.1mm power socket. I've got power supplies set up which enable LED dimming.
By the way, did you look into the Speedax mountant?

ps I like your Pleurosigma in the other thread. Thanks !

Louise
I surveyed the Georg Abele website and found equipment for making thin layers, but not any reference to Speedax or any mounting medium. Searched to and fro and found nothing.

LouiseScot
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Re: Trying out a mono camera

#11 Post by LouiseScot » Thu Apr 01, 2021 10:16 pm

Hobbyst46 wrote:
Thu Apr 01, 2021 10:11 pm
LouiseScot wrote:
Thu Apr 01, 2021 9:39 pm
Hobbyst46 wrote:
Thu Apr 01, 2021 9:20 pm
The 486nm light has improved the image. Certainly a powerful LED will be better than filtration of white light. I cannot stand the blue light in my eyes though.
Is it easy to switch the LEDs on the Swift microscope, from white to blue? On my microscope it would need fabrication of a new LED illuminator.
Thanks - I wasn't 100% sure about it. It's easier to leave the 1W Swift LED in place and just put another above it. I've already made separate 3W and 10W LEDs (mainly for DF). I just need to put the violet/blue LEDs on heatsinks and solder a short lead with a standard 5x2.1mm power socket. I've got power supplies set up which enable LED dimming.
By the way, did you look into the Speedax mountant?

ps I like your Pleurosigma in the other thread. Thanks !

Louise
I surveyed the Georg Abele website and found equipment for making thin layers, but not any reference to Speedax or any mounting medium. Searched to and fro and found nothing.
Oh ok. I thought you might have emailed him as that's how he said to contact?

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 a mono camera

#12 Post by LouiseScot » Thu Apr 01, 2021 11:23 pm

Hobbyst46 wrote:
Thu Apr 01, 2021 10:11 pm
LouiseScot wrote:
Thu Apr 01, 2021 9:39 pm
Hobbyst46 wrote:
Thu Apr 01, 2021 9:20 pm
The 486nm light has improved the image. Certainly a powerful LED will be better than filtration of white light. I cannot stand the blue light in my eyes though.
Is it easy to switch the LEDs on the Swift microscope, from white to blue? On my microscope it would need fabrication of a new LED illuminator.
Thanks - I wasn't 100% sure about it. It's easier to leave the 1W Swift LED in place and just put another above it. I've already made separate 3W and 10W LEDs (mainly for DF). I just need to put the violet/blue LEDs on heatsinks and solder a short lead with a standard 5x2.1mm power socket. I've got power supplies set up which enable LED dimming.
By the way, did you look into the Speedax mountant?

ps I like your Pleurosigma in the other thread. Thanks !

Louise
I surveyed the Georg Abele website and found equipment for making thin layers, but not any reference to Speedax or any mounting medium. Searched to and fro and found nothing.
It might not actually be available yet. Have a look here: https://www.tmg-tuebingen.de/neues-hoch ... l-speedax/

I'm not sure if there's any connection, but I've previously come across a few high refractive index resins made by doping with nanoparticles. Will be interesting to find out more.

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 a mono camera

#13 Post by LouiseScot » Fri Apr 02, 2021 4:23 pm

Got the LEDs today. Did a quick one with the violet emitter - says 410-420nm on the packing. It comes out very contrasty - perhaps because the refractive index of the Pleurax is highest at shorter wavelengths?

Also, it's a quickie one done without a proper illuminator or anything. Still, it's looking good I think.


MedNikon40xViolet_Crop00001.jpg
MedNikon40xViolet_Crop00001.jpg (133.79 KiB) Viewed 6681 times

I reused the same scale as before. I'll do some more images when I can. I'll try and find the same diatom I imaged before, else a very similar one.


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 a mono camera

#14 Post by Hobbyst46 » Fri Apr 02, 2021 8:09 pm

The UV image is indeed very good. For example, here is another example of that diatom (Petroneis humerosa) imaged under phase contrast, 100X1.3 oil objective, and blue light (~450nm). The UV image is better resolved, although there are other variables.
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LouiseScot
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Re: Trying out a mono camera

#15 Post by LouiseScot » Fri Apr 02, 2021 9:03 pm

Hobbyst46 wrote:
Fri Apr 02, 2021 8:09 pm
The UV image is indeed very good. For example, here is another example of that diatom (Petroneis humerosa) imaged under phase contrast, 100X1.3 oil objective, and blue light (~450nm). The UV image is better resolved, although there are other variables.
Hiya

Thanks! It's not actually UV, it's visible violet-blue - only a bit shorter wavelength than yours - ~415nm vs your 450nm. Your image is very good but it's clearly taken with a colour camera which won't quite reflect the true resolution of the objective. Only 1 of 4 pixels is strictly blue and although it's possible to strip out just the blue channel I still don't think you'd achieve the absolute resolution of a proper monochrome camera, but should come close. If you did strip out just the blue channel (free software called IRIS will do it : http://www.astrosurf.com/buil/iris-software.html then you'd end up with an image which is 1/4 the size of the original - if you fancy having a go. You may also be able to do it in Photoshop - I don't know). I think phase contrast in itself introduces artefacts? Though I've no direct experience of the technique.
I'm thinking the high contrast from just the short wavelength combined with the high NA, and the smaller pixels of the mono camera, are together really good for showing details of diatoms :) I've really just been investigating the capabilities of the newly acquired Nikon Fluor 40/.85 and I'm very pleased with it. I would expect to see even more detail with a 100/1.30 plan achromat - also with no CA in monochromatic light. I'll give it a go tomorrow :)

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 a mono camera

#16 Post by Hobbyst46 » Fri Apr 02, 2021 9:41 pm

LouiseScot wrote:
Thu Apr 01, 2021 11:23 pm
{ Speedax} It might not actually be available yet. Have a look here: https://www.tmg-tuebingen.de/neues-hoch ... l-speedax/
I'm not sure if there's any connection, but I've previously come across a few high refractive index resins made by doping with nanoparticles. Will be interesting to find out more.
I too believe this is the idea; Norland high RI adhesives are based on nano particles as well.

LouiseScot
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Re: Trying out a mono camera

#17 Post by LouiseScot » Fri Apr 02, 2021 10:42 pm

I just adjusted the illumination and added a lens/diffuser so it's similar to the native Swift one. Also moved the below-condenser blue filter though not sure if doing that had any effect. I centred the LED better also. So I think (I hope) this is a better image of the same target:

Med_Nikon_40x_Violet_Crop_100pc_00001.jpg
Med_Nikon_40x_Violet_Crop_100pc_00001.jpg (109.21 KiB) Viewed 6644 times


I'll definitely have a go with the 100x tomorrow. I also need to fabricate something that will give me finer control over the focusing. I'm thinking some sort of extension lever affixed to the fine focus knob. I also need to think about getting another monitor! At the moment I'm hopping between the microscope on a table and the monitor on a desk to check focus. I really need another monitor next to the microscope. I've been wanting to get a 4k monitor for several years so one of those could go in place of the desk one :)

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 a mono camera

#18 Post by MichaelG. » Sat Apr 03, 2021 5:50 am

That’s looking very good

MichaelG.
Too many 'projects'

LouiseScot
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Re: Trying out a mono camera

#19 Post by LouiseScot » Sat Apr 03, 2021 8:29 am

MichaelG. wrote:
Sat Apr 03, 2021 5:50 am
That’s looking very good

MichaelG.
Thanks! More to do!

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 a mono camera

#20 Post by MichaelG. » Sat Apr 03, 2021 9:00 am

You are already getting close to the best image here:
https://www.algaebase.org/search/specie ... s_id=34118

Keep up the great work

MichaelG.
Too many 'projects'

LouiseScot
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Re: Trying out a mono camera

#21 Post by LouiseScot » Sat Apr 03, 2021 9:10 am

MichaelG. wrote:
Sat Apr 03, 2021 9:00 am
You are already getting close to the best image here:
https://www.algaebase.org/search/specie ... s_id=34118

Keep up the great work

MichaelG.
I like to go by SEM images see: Life history and systematics of Petroneis
(Bacillariophyta), with special reference to British
waters

edited cos link didn't work :( Try this one or google the title

https://www.tandfonline.com/doi/abs/10. ... 0400024675


:)

Louise
Last edited by LouiseScot on Sat Apr 03, 2021 9:14 am, edited 1 time in total.
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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Re: Trying out a mono camera

#22 Post by MichaelG. » Sat Apr 03, 2021 9:13 am

^^^
Broken link there, Louise
Too many 'projects'

LouiseScot
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Re: Trying out a mono camera

#23 Post by LouiseScot » Sat Apr 03, 2021 9:15 am

MichaelG. wrote:
Sat Apr 03, 2021 9:13 am
^^^
Broken link there, Louise
just edited :)
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 a mono camera

#24 Post by MichaelG. » Sat Apr 03, 2021 9:21 am

Thanks

... That certainly gives you a ‘stretch target’

MichaelG.
Too many 'projects'

Hobbyst46
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Re: Trying out a mono camera

#25 Post by Hobbyst46 » Sat Apr 03, 2021 10:13 am

The linked article nicely demonstrates the structural difference between the inner and outer surfaces of the valves (ribs vs dots, say).
Can we see this difference with (home) light microscopy, rather than SEM ?

These diatoms on the slide were purposely mildly cleaned, without resort to harsh chemicals. Maybe they are complete frustules, from which the protein and lipids and pigments were removed ? In that case, one can observe only the overlaid structural patterns of both surfaces.

Alternatively, they are separated valves, that just lie there, maybe always oriented with the outer (or inner) side up...could be a preferred orientation during mounting.

I have a slide of isolated, individually selected P. humerosas, to look for surface variability.
When manually selecting diatoms for isolation, at 60X magnification under the stereoscope, differentiation between dorsal and ventral diatoms is super-hard !

LouiseScot
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Re: Trying out a mono camera

#26 Post by LouiseScot » Sat Apr 03, 2021 10:42 am

Hobbyst46 wrote:
Sat Apr 03, 2021 10:13 am
The linked article nicely demonstrates the structural difference between the inner and outer surfaces of the valves (ribs vs dots, say).
Can we see this difference with (home) light microscopy, rather than SEM ?

These diatoms on the slide were purposely mildly cleaned, without resort to harsh chemicals. Maybe they are complete frustules, from which the protein and lipids and pigments were removed ? In that case, one can observe only the overlaid structural patterns of both surfaces.

Alternatively, they are separated valves, that just lie there, maybe always oriented with the outer (or inner) side up...could be a preferred orientation during mounting.

I have a slide of isolated, individually selected P. humerosas, to look for surface variability.
When manually selecting diatoms for isolation, at 60X magnification under the stereoscope, differentiation between dorsal and ventral diatoms is super-hard !
I think SEM pics with scale bars give a better guide to the sizes of small details like pores and spacing between them - that's the only reason I make use of them. Yeah, the samples in the article have been specially prepared - either for light microscopy or SEM. Your Mediterranean slide is great for me! I wouldn't have the patience to manipulate single diatoms! My judgement and eyesight aren't so good either, so it's always a struggle to get the right focus. I just moved a monitor to be next to the Swift so that should help. I just have to figure out how to extend the display to what is effectively a third monitor on the Windows 10 system... The second one is a projector. I have a toolbar on the 'microscope' monitor but can't seem to extend the main display to it. There must be a way! Maybe I'll have to unplug the projector.

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 a mono camera

#27 Post by LouiseScot » Sat Apr 03, 2021 11:54 am

Just for fun I thought I'd try imaging some of the different layers of the Petroneis (sounds like a Harry Potter charm, doesn't it? :D ). With hindsight, I should have been a bit more methodical and done it better. Still, it's quite interesting to see some of the structural detail.

PetroneisLayerscomposite.jpg
PetroneisLayerscomposite.jpg (134.19 KiB) Viewed 6551 times


Oh, I got my extra monitor going by duplicating rather than extending. Good enough, I suppose.

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 a mono camera

#28 Post by LouiseScot » Sat Apr 03, 2021 4:18 pm

Here's a few I did at 100x. Hard to get a sharp focus. I've probably reached the limit for that 100x achro objective, at least.

Petronies_100x_Crop_415nm_014.jpg
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Petroneis_100xCrop415nm_23.jpg
Petroneis_100xCrop415nm_23.jpg (72.21 KiB) Viewed 6522 times

Petroneis_100x_crop_415nm_17.jpg
Petroneis_100x_crop_415nm_17.jpg (58.63 KiB) Viewed 6522 times

Petroneis_100x_Crop_415nm_15.jpg
Petroneis_100x_Crop_415nm_15.jpg (67.46 KiB) Viewed 6522 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 a mono camera

#29 Post by Hobbyst46 » Tue Apr 06, 2021 6:43 pm

Images 014, 023 and 017 nicely show the different structural details of the diatom. Like, ridges on the inner layer and round holes on the outer (according to the SEM images in the recently linked article).

LouiseScot
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Re: Trying out a mono camera

#30 Post by LouiseScot » Tue Apr 06, 2021 6:51 pm

Hobbyst46 wrote:
Tue Apr 06, 2021 6:43 pm
Images 014, 023 and 017 nicely show the different structural details of the diatom. Like, ridges on the inner layer and round holes on the outer (according to the SEM images in the recently linked article).
Thanks, though I couldn't seem to get sharp focus with the 100x achromat. Perhaps that could have been something to do with the short wavelength interacting with the oil? Or just the limits of the lens - I'm not sure. Maybe I need another Nikon!

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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