Naphrax vs Pleurax vs Cameras vs Me!

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LouiseScot
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Naphrax vs Pleurax vs Cameras vs Me!

#1 Post by LouiseScot » Sun Jan 10, 2021 5:10 pm

Had my first go at mounting some diatoms in Naphrax today :) . It's the same slide that I used in my 'Homegrown Diatoms' thread. As it's my first go it may be that I didn't do it so well... :( I've done a comparison with a slide using Pleurax which was very kindly donated to me. The Pleurax seems much clearer. I did manage to slightly overcook the Naphrax - might that cause a problem? My dslr images haven't generally been too good either so I also did a comparison between the Canon 1100d and Toupcam 5MP. The latter seemed slightly better? All on my Swift380T at 40x. I put the Toupcam in place of an eyepiece rather than using the tube. Again, not sure if the tube is major factor for image quality.

Pleurax slide, camera 1100d:
MedPleurax1100d40x_0002Crop50pc.jpg
MedPleurax1100d40x_0002Crop50pc.jpg (49.67 KiB) Viewed 8349 times

Pleurax slide, Toupcam:
5MP_Pleurax40x_00001_42pc.jpg
5MP_Pleurax40x_00001_42pc.jpg (58.5 KiB) Viewed 8349 times


Naphrax slide, 1100d:
Pond_Naphrax1100d40x_0004crop125pc.jpg
Pond_Naphrax1100d40x_0004crop125pc.jpg (39.62 KiB) Viewed 8349 times

Naphrax slide, Toupcam:
5MP_Naphrax40x_Crop_01.jpg
5MP_Naphrax40x_Crop_01.jpg (55.89 KiB) Viewed 8349 times
The Pleurax slide seems much better to me. My dslr pics always seem to come out not very clear - no sharp focus. I've been using a Chinese 'dslr adapter' for the trinocular attachment (which has an inbuilt lens) - not sure if the images through that are as good as they might be.
Any thoughts, suggestions?

Thanks
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: Naphrax vs Pleurax vs Cameras vs Me!

#2 Post by MicroBob » Sun Jan 10, 2021 7:37 pm

Hi Louise,
nice to see that you have started to try cleaning and mountig diatoms yourself! The results are actually not bad and there are still some points to check for possible improvement left. Small diatoms are at the border of what a light microscope can resolve in detail, so it is common to end up with a rough idea of the shape but not much more.

Here some hints that might lead to further improvement:

-Diatom slides should be made upside down. This leads to a slide where the diatoms are close to the cover slip. Place a cleaned round cover slip on a nut (like M6-nut), apply diatom material in a drop of water of ethanol (or dry when extremely clean), let dry fully.

-Naphrax is dissolved in toluene and a drop of toluene should be applied to the diatoms befor the naphrax is applied. Then the cover slip is put onto a hot plate at 100°C to let the toluene evaporate, only then the Naphrax reaches it's full refracrive index. Don't overheat as Naphrax has a tendency o decay which might be provoked by overheating.

-Thorougly clean a slide, they are hygroscopic, you don't want to trap water drops, warm the slide a bit

-Pick up the cover slip with diatoms in Naphrax and let it cool in upside down position. This can also be done with the cover slip on a nut.

- For great photos it is important to use a camera with electronic first shutter curtain function. This means that from live view the shutter doesn't move before a photo is taken as this shutter movement intoduces some vibration that leads to blurred images. Many Canons have this feature, but not all - you should check this and activate the feature if available. if your camera doesn't have EFSC you can try to dampen the shock by supporting the camera with a piece of wood or a tube against the wall on bottom or top as the shutter moves up-down.

-Diatoms are transparent so you can play around in post editing with the b/w conversion. I often separate the image in RGB channels with GIMP and use the green channel.

- Use a 100:1 oil immersion objective if you have one. It offers twice the resolution of a 40:1. For some objects this is not worth the the effort but with diatoms we need every bit of resolution we can get.

- Cleaning can be seen as a multi step affair. Next to the organic content in the diatom there is other organic matter, silt and calerous matter that could be removed to further improve the slide. Silt can be removed by sieving throught e.g. a 25µ stainless sieve (material affordable on ebay). Calcerous matter cna be removed with concentrated vinegar or citric acid or HCl..

- Cleaning by incineration is an attractive alternative: Place a 4mm stainless steel plate on an alcohol burner, cover slip on there, drop of diatom rich water sample on covre slip, heat from below. Not too hot, otherwise the cover slip and diatoms deform. Then the trick: Apply the flame of a blue flame-cigarette-lighter on the diatoms on the cover slip on the steel plate. When you find the right amound of heating you get a nice dirty slide with fairly clean diatoms - and colonies stay together! Attatched is a photo of a fragellaria colony making a roll.

I would be happy to read how you get ahead with this project!

Bob
Bob
Attachments
Fragelaria Kolonie macht eine Rolle IMG_20200701_154109-04 bearbeitet 1024.jpg
Fragelaria Kolonie macht eine Rolle IMG_20200701_154109-04 bearbeitet 1024.jpg (48.82 KiB) Viewed 8332 times

Hobbyst46
Posts: 4277
Joined: Mon Aug 21, 2017 9:02 pm

Re: Naphrax vs Pleurax vs Cameras vs Me!

#3 Post by Hobbyst46 » Sun Jan 10, 2021 9:09 pm

LouiseScot wrote:
Sun Jan 10, 2021 5:10 pm
The Pleurax slide seems much better to me. My dslr pics always seem to come out not very clear - no sharp focus. I've been using a Chinese 'dslr adapter' for the trinocular attachment (which has an inbuilt lens) - not sure if the images through that are as good as they might be.
Any thoughts, suggestions?
Compliments on the Naphrax diatom slide ! working with diatoms is challenging, but rewarding !
Although the photo only shows a fraction of the FOV, the slide appears fine: the diatom looks clean, there are no air bubbles and no debris. There is no air bubble within the diatom.
Naphrax should be at least as good as Pleurax - their RI's are similar when the solvent (toluene and isopropanol, respectively) is removed.

I am not sure that the Naphrax image is worse than the Pleurax image, resolution-wise. The dimensions of the diatoms are not shown.
It might be that the mountant layer is thicker than optimal, yet in my experience, this only shows up in diatoms whose stria spacing is extremely fine. So I believe that it is not an issue for these slides.
Does the Naphrax diatom appear sharper and better resolved in eye-view ?

LouiseScot
Posts: 1167
Joined: Tue Jul 07, 2020 1:51 pm
Location: Scotland

Re: Naphrax vs Pleurax vs Cameras vs Me!

#4 Post by LouiseScot » Sun Jan 10, 2021 10:07 pm

MicroBob wrote:
Sun Jan 10, 2021 7:37 pm
Hi Louise,
nice to see that you have started to try cleaning and mountig diatoms yourself! The results are actually not bad and there are still some points to check for possible improvement left. Small diatoms are at the border of what a light microscope can resolve in detail, so it is common to end up with a rough idea of the shape but not much more.

Here some hints that might lead to further improvement:

-Diatom slides should be made upside down. This leads to a slide where the diatoms are close to the cover slip. Place a cleaned round cover slip on a nut (like M6-nut), apply diatom material in a drop of water of ethanol (or dry when extremely clean), let dry fully.

-Naphrax is dissolved in toluene and a drop of toluene should be applied to the diatoms befor the naphrax is applied. Then the cover slip is put onto a hot plate at 100°C to let the toluene evaporate, only then the Naphrax reaches it's full refracrive index. Don't overheat as Naphrax has a tendency o decay which might be provoked by overheating.

-Thorougly clean a slide, they are hygroscopic, you don't want to trap water drops, warm the slide a bit

-Pick up the cover slip with diatoms in Naphrax and let it cool in upside down position. This can also be done with the cover slip on a nut.

- For great photos it is important to use a camera with electronic first shutter curtain function. This means that from live view the shutter doesn't move before a photo is taken as this shutter movement intoduces some vibration that leads to blurred images. Many Canons have this feature, but not all - you should check this and activate the feature if available. if your camera doesn't have EFSC you can try to dampen the shock by supporting the camera with a piece of wood or a tube against the wall on bottom or top as the shutter moves up-down.

-Diatoms are transparent so you can play around in post editing with the b/w conversion. I often separate the image in RGB channels with GIMP and use the green channel.

- Use a 100:1 oil immersion objective if you have one. It offers twice the resolution of a 40:1. For some objects this is not worth the the effort but with diatoms we need every bit of resolution we can get.

- Cleaning can be seen as a multi step affair. Next to the organic content in the diatom there is other organic matter, silt and calerous matter that could be removed to further improve the slide. Silt can be removed by sieving throught e.g. a 25µ stainless sieve (material affordable on ebay). Calcerous matter cna be removed with concentrated vinegar or citric acid or HCl..

- Cleaning by incineration is an attractive alternative: Place a 4mm stainless steel plate on an alcohol burner, cover slip on there, drop of diatom rich water sample on covre slip, heat from below. Not too hot, otherwise the cover slip and diatoms deform. Then the trick: Apply the flame of a blue flame-cigarette-lighter on the diatoms on the cover slip on the steel plate. When you find the right amound of heating you get a nice dirty slide with fairly clean diatoms - and colonies stay together! Attatched is a photo of a fragellaria colony making a roll.

I would be happy to read how you get ahead with this project!

Bob
Bob
Thanks for all the advice, Bob. Did you see my previous post on 'homegrown diatoms' from the other day? I explained how I prepared the diatoms in situ - i.e. straight from jar to slide. Yeah, I know I should really do the prep on the cover slip but it's too difficult for me really. I don't have the dexterity or the eyesight, I'm afraid :( I will have a go next time but I'm not optimistic... All my photomicrography has been done in Canon live view (or with the 5MP Toupcam). The above dslr pics are taken with a 1100d. I also tried a 550d but that didn't seem so good - don't know why. (In general, I may be photographically over-enlarging what are small diatoms.) However, I recently purchased a Fuji X-T30 mirrorless but I've yet to try it on a microscope. I'll certainly give a 100x oil immersion a go. When I did the Naphrax I put a drop of Limonene on the slide first as I didn't have any toluene but will get some. Unfortunately, in my domestic situation, I don't have lab facilities but I accept such limitations and do what I can! I have just got some Trichlor which I thought may work better than 'Domestos' for cleaning the diatoms. I'll try it next time. I have a sample in a vial which I'll try treating. Oh, a while ago, I previously tried imaging with a mono camera but I couldn't see any difference in resolution or clarity.

Anyway, thanks again

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
Posts: 4277
Joined: Mon Aug 21, 2017 9:02 pm

Re: Naphrax vs Pleurax vs Cameras vs Me!

#5 Post by Hobbyst46 » Sun Jan 10, 2021 10:26 pm

Nice choice, limonene as substitute for toluene. Likely, it will dissolve Naphrax well (my guess). Its disadvantage for Naphrax mounting is its high boiling point of 176C. Hence the slide must be heated long enough to remove the limonene, otherwise the RI will not reach 1.7. Of course heating the slide to 170-180C will cause rapid evaporation but perhaps Naphrax is not stable at such high temp's. So, may I suggest the following:
1. The slide that bears the diatom suspension drop (taken from the jar) should be left on the shelf to dry out completely, say for 48 hours.
2. After limonene and Naphrax are added and the coverslip is in place, the slide should be heated to about 100C ( or as near to it as possible) and left at this temperature for several hours.
These are suggested based on intuition, not personal experience with Naphrax.

LouiseScot
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Location: Scotland

Re: Naphrax vs Pleurax vs Cameras vs Me!

#6 Post by LouiseScot » Sun Jan 10, 2021 10:30 pm

Hobbyst46 wrote:
Sun Jan 10, 2021 9:09 pm
LouiseScot wrote:
Sun Jan 10, 2021 5:10 pm
The Pleurax slide seems much better to me. My dslr pics always seem to come out not very clear - no sharp focus. I've been using a Chinese 'dslr adapter' for the trinocular attachment (which has an inbuilt lens) - not sure if the images through that are as good as they might be.
Any thoughts, suggestions?
Compliments on the Naphrax diatom slide ! working with diatoms is challenging, but rewarding !
Although the photo only shows a fraction of the FOV, the slide appears fine: the diatom looks clean, there are no air bubbles and no debris. There is no air bubble within the diatom.
Naphrax should be at least as good as Pleurax - their RI's are similar when the solvent (toluene and isopropanol, respectively) is removed.

I am not sure that the Naphrax image is worse than the Pleurax image, resolution-wise. The dimensions of the diatoms are not shown.
It might be that the mountant layer is thicker than optimal, yet in my experience, this only shows up in diatoms whose stria spacing is extremely fine. So I believe that it is not an issue for these slides.
Does the Naphrax diatom appear sharper and better resolved in eye-view ?
The Naphrax doesn't seem so clear to me. I can't seem to get the striae resolution and clarity that the Pleurax has.. Yes, the diatoms do appear sharper and clearer via the oculars - but a lot smaller, of course. I did try taking an image with a smartphone over an eyepiece but wasn't very successful. Do you think it's the photo enlargement that's the issue? Other people seem to be able to get much better images than I ever get though it's all a work in progress for me. I have tried the 60x plan achro but it's still a bit blurry. I'll give a 100x a go tomorrow. The Naphrax seemed quite thin and I pressed down on the cover slip but I've no experience so it's hard to judge. Hopefully, things will improve with practice! When the weather warms up I'll go on the hunt for some larger species.
Here's what I got with the phone and the Pleurax slide. (I'm so glad I have the Pleurax slides as a reference! )
PleuraxPhone50pc.jpg
PleuraxPhone50pc.jpg (48 KiB) Viewed 8302 times
Cheers

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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Location: Scotland

Re: Naphrax vs Pleurax vs Cameras vs Me!

#7 Post by LouiseScot » Sun Jan 10, 2021 10:38 pm

Hobbyst46 wrote:
Sun Jan 10, 2021 10:26 pm
Nice choice, limonene as substitute for toluene. Likely, it will dissolve Naphrax well (my guess). Its disadvantage for Naphrax mounting is its high boiling point of 176C. Hence the slide must be heated long enough to remove the limonene, otherwise the RI will not reach 1.7. Of course heating the slide to 170-180C will cause rapid evaporation but perhaps Naphrax is not stable at such high temp's. So, may I suggest the following:
1. The slide that bears the diatom suspension drop (taken from the jar) should be left on the shelf to dry out completely, say for 48 hours.
2. After limonene and Naphrax are added and the coverslip is in place, the slide should be heated to about 100C ( or as near to it as possible) and left at this temperature for several hours.
These are suggested based on intuition, not personal experience with Naphrax.
Thanks. I'll order some Toluene tomorrow. The slide was very dry as I'd made it several days before. As I mentioned, I did wonder if I'd overcooked the Naphrax - it had turned slightly brown... (the cooker hotplate got hotter than expected!) I had done a test with the Limonene and it evaporated quickly on the hot plate at ~100 deg. I'd also wicked off most of it on the Naphrax slide so shouldn't have been an issue but I'm not sure.
Thanks

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
Posts: 1167
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Location: Scotland

Re: Naphrax vs Pleurax vs Cameras vs Me!

#8 Post by LouiseScot » Mon Jan 11, 2021 12:12 pm

I stayed up late last night and had a go with the Swift 100x. I continue to be impressed with the optics of this scope which only cost £216 from Amazon!
Anyway, here are some taken with the Canon 1100d. I'll post Toupcam pics separately.

Naphrax:
Naphrax1100d_100x_2crop.jpg
Naphrax1100d_100x_2crop.jpg (70.02 KiB) Viewed 8254 times
Naphrax1100d_100x_5_crop.jpg
Naphrax1100d_100x_5_crop.jpg (68.97 KiB) Viewed 8254 times
Naphrax1100d_100x_6crop.jpg
Naphrax1100d_100x_6crop.jpg (56.86 KiB) Viewed 8254 times

Pleurax:
Pleurax1100d_100x_1_crop_36pc.jpg
Pleurax1100d_100x_1_crop_36pc.jpg (88.18 KiB) Viewed 8254 times
Pleurax1100d_100x_3crop41pc.jpg
Pleurax1100d_100x_3crop41pc.jpg (66.32 KiB) Viewed 8254 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

LouiseScot
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#9 Post by LouiseScot » Mon Jan 11, 2021 12:15 pm

5MP Toupcam pics, 100x
5MP_Naphrax_100x_00001crop.jpg
5MP_Naphrax_100x_00001crop.jpg (59.04 KiB) Viewed 8253 times
5MP_Naphrax_100x_00015_crop.jpg
5MP_Naphrax_100x_00015_crop.jpg (69.06 KiB) Viewed 8253 times
5MP_Pleurax_100x_1_00001.jpg
5MP_Pleurax_100x_1_00001.jpg (70.34 KiB) Viewed 8253 times
5MP_Pleurax_100x_1_00002Crop.jpg
5MP_Pleurax_100x_1_00002Crop.jpg (58.57 KiB) Viewed 8253 times

I think the Toupcam images are very good but very small fov at 100x!

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: Naphrax vs Pleurax vs Cameras vs Me!

#10 Post by Hobbyst46 » Mon Jan 11, 2021 1:38 pm

Certainly even better images !
improved sharpness, higher resolution and less CA, supporting the hypothesis that the Naphrax slide is OK, and the images can be improved via the photographic gear.
I think that the Canon 1100D altogether yields slightly better pictures than the Toupcam.
Are you using a 0.5X or other reducing lens with the Toupcam ?

LouiseScot
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#11 Post by LouiseScot » Mon Jan 11, 2021 1:54 pm

Hobbyst46 wrote:
Mon Jan 11, 2021 1:38 pm
Certainly even better images !
improved sharpness, higher resolution and less CA, supporting the hypothesis that the Naphrax slide is OK, and the images can be improved via the photographic gear.
I think that the Canon 1100D altogether yields slightly better pictures than the Toupcam.
Are you using a 0.5X or other reducing lens with the Toupcam ?
Hiya

Thanks! I still think the Pleurax is better... The Toupcam is just put in place of an eyepiece. I'm a bit suspect of the Chinese dslr adapter I've been using so I'm just in the process of making an alternative adapter based on an astronomy 2" eyepiece :) I have to 3d print a part for that.

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: Naphrax vs Pleurax vs Cameras vs Me!

#12 Post by MicroBob » Mon Jan 11, 2021 3:36 pm

Hi Louise,
using the 100:1 has brought things forward a lot! I think the setup is fine like this and there is only a narrow margin for further improvements in the raw image. I have taken the blue channel, worked a bit on the histogram and did a bit of unsharp masking and the striae are visible very well.
At this point of image quality the illumination becomes more important, correct setup of the condenser and setup of oblique or circular oblique lighting. Perhaps there is something to gain.
If you have a condenser with numeric aperture over 1.0 you might try to use dem. water or immersion oil between top lens and slide.

As far as I can see the 1100D ahould have EFSC from live view though it is not really documented. So the camera itself is no limit. The typical camera adaptation for a finity microscope would be to place the APS-Ccamera with a flatly built camera lens/objective of about 40mm focal length on to of a high eyepoint eyepiece that fits the correction needs of the microscope objectives. Usually the higher power microscope objectives have som residual color error that is corrected by the eyepiece in mor expensive microscopes. For your microscope it will be a question of trial and error to find the best eyepiece for the camera adaptation. If it doesn't fit well colour fringes will appear towards the border of the image.

Bob
Attachments
Louisediatom.jpg
Louisediatom.jpg (80.37 KiB) Viewed 8231 times

LouiseScot
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#13 Post by LouiseScot » Mon Jan 11, 2021 4:02 pm

MicroBob wrote:
Mon Jan 11, 2021 3:36 pm
Hi Louise,
using the 100:1 has brought things forward a lot! I think the setup is fine like this and there is only a narrow margin for further improvements in the raw image. I have taken the blue channel, worked a bit on the histogram and did a bit of unsharp masking and the striae are visible very well.
At this point of image quality the illumination becomes more important, correct setup of the condenser and setup of oblique or circular oblique lighting. Perhaps there is something to gain.
If you have a condenser with numeric aperture over 1.0 you might try to use dem. water or immersion oil between top lens and slide.

As far as I can see the 1100D ahould have EFSC from live view though it is not really documented. So the camera itself is no limit. The typical camera adaptation for a finity microscope would be to place the APS-Ccamera with a flatly built camera lens/objective of about 40mm focal length on to of a high eyepoint eyepiece that fits the correction needs of the microscope objectives. Usually the higher power microscope objectives have som residual color error that is corrected by the eyepiece in mor expensive microscopes. For your microscope it will be a question of trial and error to find the best eyepiece for the camera adaptation. If it doesn't fit well colour fringes will appear towards the border of the image.

Bob
Hi again, Bob

Thanks :) The one you've enhanced was taken with the Toupcam. If one wanted to dismiss osc colour it would be better to use a mono camera with filter(s) since only 1 in 4 pixels in a colour camera is blue. I believe my illumination is set up correctly but it's always worth double checking! Sometimes I use an iris diaphragm over the LED light source to either simulate Kohler or to provide oblique lighting. The 100x has a standard NA of 1.25 so I try and match with that. I don't think the Swift380T has any correction requirements - it's pretty basic. It has a lens in the trinocular port and I've set things up to make the dslr parfocal with the eyepieces. As mentioned above I'm just making a dslr adapter to connect a 28mm 2" telescope eyepiece (I just happen to have one which I don't really use) in place of the current Chinese dslr adapter. Also, in the next few days I'll try out the Fuji X-T30 in place of the Canon dslrs. It's really destined for my focus rail microscope but I like experimenting :)
Thanks for all your feedback!

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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Location: Scotland

Re: Naphrax vs Pleurax vs Cameras vs Me!

#14 Post by LouiseScot » Tue Jan 12, 2021 12:30 am

Just did a quick test of my DIY dslr adapter at 40x on the Canon 1100d. Seems to work ok though I haven't done a close comparison. I'll try it at 100x tomorrow. Oh, I think I found the problem I had with the Canon 550d. It seems to remember the F stop setting from whatever lens was on it previously. It keeps it until you put another auto lens on, apparently. I haven't confirmed it but it would explain some strange behaviour!


Pleurax1100d_40x_EP2crop70pc - Copy.jpg
Pleurax1100d_40x_EP2crop70pc - Copy.jpg (92.85 KiB) Viewed 8199 times
Pleurax1100d_40x_EP3crop31pc - Copy.jpg
Pleurax1100d_40x_EP3crop31pc - Copy.jpg (118.03 KiB) Viewed 8199 times

I'll put some details of the adapter in the camera systems and imaging forum :)

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

KitKestrel
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#15 Post by KitKestrel » Tue Feb 02, 2021 4:49 am

Great thread! Very inspiring to this one who is new to diatom arts :) Does anyone happen to know if there is a supplier anywhere for Pleurax? I hope to try hobby46's recipe for homemade mountant, but as a backup I thought I might try Pleurax as well, as it seems to be the least toxic to use of the commercial brands, and I don't have access to a lab.

Thanks!
Kit.

MicroBob
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#16 Post by MicroBob » Tue Feb 02, 2021 6:38 am

Hi Kit,
Michel sells Pleurax but is taking a break at the moment: https://www.diatoms.nl/index.php

Bob

photomicro
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#17 Post by photomicro » Tue Feb 02, 2021 3:23 pm

KitKestrel wrote:
Tue Feb 02, 2021 4:49 am
Great thread! Very inspiring to this one who is new to diatom arts :) Does anyone happen to know if there is a supplier anywhere for Pleurax? I hope to try hobby46's recipe for homemade mountant, but as a backup I thought I might try Pleurax as well, as it seems to be the least toxic to use of the commercial brands, and I don't have access to a lab.

Thanks!
Kit.
If you only want a small amount to try, and in Europe, I could supply some.

Send a PM.

Mike

KitKestrel
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#18 Post by KitKestrel » Tue Feb 02, 2021 7:08 pm

Hi Mike,

Thanks so much for the offer! I do only need a little bit, but I am unfortunately in the US, which appears to be a desert of mountant... I have to believe that there are amateur diatom enthusiasts here who don't have access to a lab. I wonder what they all do...

In any case, your offer is appreciated :-)

Kit.

MicroBob
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#19 Post by MicroBob » Tue Feb 02, 2021 8:39 pm

Hi Kit,
if you can't coax Mike into sending you some Pleurax, risking a holiday in Guantamo bay :D , you might try these simple mountants:

- air
- water

For good visiblility a difference of the refractive index of 0,1 is recommended, diatoms are around 1,45. So these mediums will do both quite well.

Two limitations:
- with air no more than n.a. 1,0 is available due to total reflection between coverslip and mounting air
- with both no perfectly fixed frustules, they might move or collect in one corner when holding the slide vertical

Enclosed a diatom in dried out salt water. Cover slip held on with epoxy from the holiday tool kit. I was about to throw awa the slide but then decided to have a loser look.

Bob
Attachments
Diatomee Centrales Salzwasser 1024 DSC_5402.jpg
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LouiseScot
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#20 Post by LouiseScot » Tue Feb 02, 2021 9:16 pm

Yes, air works

viewtopic.php?f=6&t=11535

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: Naphrax vs Pleurax vs Cameras vs Me!

#21 Post by Hobbyst46 » Tue Feb 02, 2021 9:18 pm

Bob, the diatom is great, and the overall view is of a solar-roof alien spaceship landing in the Alps.

KitKestrel
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#22 Post by KitKestrel » Wed Feb 03, 2021 4:18 pm

:D

I've used water in flat well slides and it works reasonably well but wasn't trying to make a permanent slide. I'll try both air and water. I don't mind if things shift around - they'll be for my use, not students or anyone else so the risk of damage is slight.

My first challenge is working out how to prepare the samples without noxious chemicals - no lab, no hood. Am experimenting with diatomite, so more clay, less organic matter. Planning to concentrate vinegar via freezing and use that for the clay, and if necessary use H2O2 (hair developer) for organic matter. The big question is - in what fraction will the diatoms appear!?

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Re: Naphrax vs Pleurax vs Cameras vs Me!

#23 Post by MicroBob » Wed Feb 03, 2021 4:54 pm

These well slides have a stong impact on image quality - you have a water lens in the light path. I can't remember how I made the slide. It was mediterranean sea water with about 4% salt. The cover slip is held on with 5-minute expoxy, but I don't know wheteher I gave the water time to evaporate before sealing the slide.

Here is a presentation on diatom cleaning methods for the kitchen lab: http://www.mikrohamburg.de/Programm/Pro ... 170917.pdf
In most cases a multi step method will provide the best results.
For work on diatomite clay you might use a peltier element to get quick freeze-thaw-cycles.
Silt might be removed by washing in a very fine (e.g. 25µ) sieve, loosing most of the smaller diatoms. Calcerous matter with concentrated vinegar. Letting settle the diatoms in champagne glasses also helps to divide dirt from frustules, but at the risk to discard the wrong part. And bid frustules sink as fast as small sand grains...
Perhaps your frustules are already clean enough then.

In case you try to process fresh diatoms on day:
For cleaning the frustules heat is an option. Too hot an too many frustules will deform. In that presentation I wrote 550 and 900 °C. Today I would stay below 450 °C.
Advantage of the incineration method is that colonies stay together.
Other options are cold sodiumhypochloride (bleach) or hot ammoniumpersulfate (used for circuit board etchning)

Bob

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Re: Naphrax vs Pleurax vs Cameras vs Me!

#24 Post by KitKestrel » Wed Feb 03, 2021 6:22 pm

Thanks much - I experiment with those suggestions.

Kit.

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Re: Naphrax vs Pleurax vs Cameras vs Me!

#25 Post by KitKestrel » Fri Feb 05, 2021 4:35 pm

@microbob Your note about the peltier element - I wasn't familiar with these, so looked them up. I am not sure how I would apply the examples I found to this application. Could you explain a little more how to set it up? Thanks!

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Re: Naphrax vs Pleurax vs Cameras vs Me!

#26 Post by MicroBob » Fri Feb 05, 2021 5:00 pm

Hi Kit,
the idea is to repeatedly freeze and thaw the soaked block of diatom material. This can be done with a normal household freezer, but it is time consuming. With a peltier element and a time switch the process can be rund automatically and in faster cycle times. The peltier element is a semi condutor that moves heat from one side to the other. So you transport heat away from your sample, sitting in water in a sheet metal container and freeze it.
Two things are important:
- You have to get rid of the heat on the warm side, otherwise the peltier element is destroyed quickly. A CPU cooler can do this nicely
- No condensation may reach into the peltier element as the ice build up could bust the element. So a silicone seal is necessary

Here a picture of my peltier element.

Bob
Attachments
Peltier-Kühler.jpg
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KitKestrel
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#27 Post by KitKestrel » Fri Feb 05, 2021 6:30 pm

MicroBob wrote:
Fri Feb 05, 2021 5:00 pm
Hi Kit,
the idea is to repeatedly freeze and thaw the soaked block of diatom material. This can be done with a normal household freezer, but it is time consuming. With a peltier element and a time switch the process can be rund automatically and in faster cycle times. The peltier element is a semi condutor that moves heat from one side to the other. So you transport heat away from your sample, sitting in water in a sheet metal container and freeze it.
Two things are important:
- You have to get rid of the heat on the warm side, otherwise the peltier element is destroyed quickly. A CPU cooler can do this nicely
- No condensation may reach into the peltier element as the ice build up could bust the element. So a silicone seal is necessary
Cool! So what are the wires plugged into out of the picture on the right hand side? i.e., what is the power source?
Do the water and diatom blocks go in the metal basin you show, or in a container in the metal basin?
How does the thawing happen - just wait for the room air to warm it?
Is the silicone seal painted over the whole peltier element? I'm assuming the comment about condensation means water coming off the outside of the metal basin.
Thanks! Kit.

MicroBob
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#28 Post by MicroBob » Fri Feb 05, 2021 6:45 pm

So what are the wires plugged into out of the picture on the right hand side? i.e., what is the power source?
Peltiers are usually powered with low voltages, here up to 12V and 3 A or so. The CPU cooler runs on the same voltage. I use a small lab power supply for my LEDs and it is used here too.

Do the water and diatom blocks go in the metal basin you show, or in a container in the metal basin?
That is a paraffin embedding mold for standard histology frames. You can use anything that has good heat transport and doesn't break when the water freezes.

How does the thawing happen - just wait for the room air to warm it?
This would be most simple and a bit slow. You could increase the heating up with a halogen desk lamp, placed over the metal basin. But as soon as this really increases thawing speed, it will stop the peltier from freezing the block. So you would have to use two time switches that switch at opposite times.
Is the silicone seal painted over the whole peltier element? I'm assuming the comment about condensation means water coming off the outside of the metal basin.
A Peltier element has closed top and bottom and is somewhat open from the sides. This stops air humidity from thawing and freezing there.

The top and bottom of the Peltier should be attatched to cooler and top plate with thermal conductive compound used for CPUs etc..

By stacking two Peltiers one can increase the maximum difference between room temperature and top plate temperature.

KitKestrel
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#29 Post by KitKestrel » Fri Feb 05, 2021 8:02 pm

:D :D :D

rkhichi
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Re: Naphrax vs Pleurax vs Cameras vs Me!

#30 Post by rkhichi » Fri Jan 21, 2022 10:54 am

Please provide a cleaning procedure and slide preparation for diatoms in marine sediment.

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