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Arachnoidiscus ehrenbergii

 
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sushidelic



Joined: 10 Jun 2016
Posts: 124

PostPosted: Sat Jan 14, 2017 2:03 pm    Post subject: Arachnoidiscus ehrenbergii Reply with quote

Hello,

here's another diatom I like a lot. Extern lighting and this time the SPlan Apo 60x/1.4.
The most interesting thing was, when I examined the center closely, it was revealing some dots with defined lit and dark side, that each measured around 300nm in diameter. Should be close to the resolution limit of classic light microscopy?





Best regards,
Michael

PS: I hope with my recent post I didn't kill my whole quota of pics for january, and it's okay to post another one...
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zzffnn



Joined: 22 May 2014
Posts: 1400
Location: Texas USA

PostPosted: Sat Jan 14, 2017 3:20 pm    Post subject: Reply with quote

Very nice image!

But exactly how did you provide "external light" for an oil immersion 60x NA 1.4 objective? Isn't the working distance of than objective less than 0.3 mm?

I found a working distance of 2mm very challenging for external light. I cannot envision how external light can pass through WD of less than 0.3mm. Are you using fiber optic coupled to microscope slide? Even that, you would need to stop down objective to much less than NA 1.4 (to around NA 1.1).

Would you please kindly show a photo of your light/microscope arrangement?
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Selling my inverted phase contrast 20x objective for petri dish peeking
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Pau
Site Admin


Joined: 20 Jan 2010
Posts: 3894
Location: Valencia, Spain

PostPosted: Sat Jan 14, 2017 3:40 pm    Post subject: Reply with quote

Magnificent!

I'm also interested in your illumination technique, are you illuminating the immersion oil or the slide glass itself?

PS: don't worry, they are just two images (actually one) in a day
Wink
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Pau
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sushidelic



Joined: 10 Jun 2016
Posts: 124

PostPosted: Sun Jan 15, 2017 8:40 am    Post subject: Reply with quote

Just two quick pics from the setup (weekend is kids' time...).
I'll explain it a bit more later!




Best regards,
Michael
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Cactusdave



Joined: 09 Jun 2009
Posts: 1522
Location: Bromley, Kent, UK

PostPosted: Sun Jan 15, 2017 12:06 pm    Post subject: Reply with quote

Brilliantly simple. Very Happy Your images are brilliant too BTW. Very Happy I have done something similar with the diffusing cylinder and incident lights, I used a section cut from a breakfast health drink bottle ('Actimel' in the UK). But this was for a low power subject. I'm sure other forum members have done something similar. What I'm astounded by is that it gives enough light for high power objectives and even oil immersion. I have to try it. What sort of exposure times are you getting with the X63 objective for example? Also what camera are you using?

I'm also very impressed with how clean your stacks are. Whether it's another benefit of the lighting, I don't know, but I often find problems of halos and smearing (I call it 'melting' because that's what it looks like) of detail with stacking high power images of high contrast subjects like diatoms and radiolaria.

The example radiolaria and diatoms you have photographed are really nice examples as well. Are these mounts you have made yourself or carefully selected examples from commercial prepared mounts?
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Leitz Ortholux 1, Zeiss standard, Nikon Diaphot inverted, Canon photographic gear
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Pau
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Joined: 20 Jan 2010
Posts: 3894
Location: Valencia, Spain

PostPosted: Sun Jan 15, 2017 12:26 pm    Post subject: Reply with quote

I concur with all what Dave says. So simple and effective and nobody has published it before Shocked
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Pau
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sushidelic



Joined: 10 Jun 2016
Posts: 124

PostPosted: Sun Jan 15, 2017 1:05 pm    Post subject: Reply with quote

I actually have no clue, where the light passes in the end. Sure it goes through the slide, refracted, reflected on the condensor, oil, whereever... It's true, that even with only the 10x apo I'm unable to shed light on top of the eye of a daphnia, so it is basically a very oblique illumination, but due to the diffusor, it is still somewhat soft light. That helps revealing detail and for stacking Smile.

The diatoms are from a bunch of "stray" slides I bought on ebay, among those random diatoms you always find some that are well oriented and in good shape. The guy who made them embedded them in zrax, with literally no dirt at all.

Camera was a Nikon D800 with an NFK 1.67, and expoure times where roughly 5-10 seconds with ISO 100. Those long exposure times are annoying as shooting stacks takes ages, but at the same time, shutter shock is no issue at all.

Best regards,
Michael
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zzffnn



Joined: 22 May 2014
Posts: 1400
Location: Texas USA

PostPosted: Sun Jan 15, 2017 1:33 pm    Post subject: Reply with quote

Thank you. I have never tried 5-10 second exposure. That explained most of it.
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carlos.uruguay



Joined: 23 Feb 2012
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Location: Uruguay - Montevideo - America del Sur

PostPosted: Sun Jan 15, 2017 1:41 pm    Post subject: Reply with quote

SUPER!!!!
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Jacek



Joined: 02 Oct 2011
Posts: 4625
Location: Poland

PostPosted: Sun Jan 15, 2017 2:00 pm    Post subject: Reply with quote

Super !
What you have on a glass slide to obtain black background?
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Olympusman



Joined: 15 Jan 2012
Posts: 3330

PostPosted: Sun Jan 15, 2017 2:17 pm    Post subject: Diatom Reply with quote

Nice image interesting ligthing technique.

Mike
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sushidelic



Joined: 10 Jun 2016
Posts: 124

PostPosted: Mon Jan 16, 2017 2:27 am    Post subject: Reply with quote

Thanks a lot for you replies!

With an NA of 1.4, it was impossible to obtain a black background. Unfortunately, the 60x Apo doesn't have an diafragm, so i had to use the full resolution, and the source image looks kinda weird DIC style. Fortunately, most specimen have a clear border, so with careful masking the black BG is added in Photoshop. If you use apertures below 1, it can look similar to DF with a near black BG right on the source image.

Best regards,
Michael
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billben74



Joined: 09 Apr 2016
Posts: 130
Location: Cambridge, UK

PostPosted: Wed Jan 18, 2017 3:11 pm    Post subject: Reply with quote

Fantastic shot. The lighting works very well. I love it.
Really interesting technique. As I am sure many will, this is definately on my to do list.
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abpho



Joined: 17 Aug 2011
Posts: 1423
Location: Earth

PostPosted: Wed Feb 01, 2017 7:42 pm    Post subject: Reply with quote

Wow. Amazing. Looks like a maze. Very hypnotic. Congratulations on making the picture of the month too. Applause
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sushidelic



Joined: 10 Jun 2016
Posts: 124

PostPosted: Fri Feb 03, 2017 1:52 am    Post subject: Reply with quote

Thanks a lot! 1000x thanks a lot!

When i decided to get the Vanox back in october last year, I wasn't really sure wether this is just an expensive crazy idea out of a midlife mood.
But now i have found something that really adds to my life, and the results are encouraging me to keep experimenting and discussing, and shooting Smile.

Well, thanks again!

Best regards,
Michael
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