Algae galore

Images made through a microscope. All subject types.

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ModelZ
Posts: 38
Joined: Sun Nov 01, 2020 4:34 am
Location: Northern Europe

Algae galore

Post by ModelZ »

Trying to capture some green algae pics has been both good fun & challenging. Some species are essentially flat and yield e.g. to DIC pretty well, but some are strikingly 3-d and need more to render in a pleasing way.
Case in point is Oedogonium & Spirogyra in the first pic, both fresh water green algae. The former doesn't have a distinct internal structure and DIC works ok. On the other hand for Spirogyra (tube diameter 30-40 micrometers) such pics tend to appear flattish even when stacked so deep that spatial relations become logically clear. A couple more of those pics below. Surface detail turn out well in the DIC relief but the chloroplast helices less so. Polarized light pic, while colorful (with 1 Red) isn't much more illuminating structurewise (4.).
So I turned on UV & filled in with visible. Some fine tuning required since the Spirogyra shell does not fluoresce. So plenty experimenting and pic shuffling in post. But in my mind this approach serves Spirogyra type algae reasonably well.
Pics with PZO-DIC & Zetopan Epi with Zeiss Apos and Lomo Lumams.
No chemistry, just light & filters. All medium stacks.

Cheers,
-Karl
Attachments
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d1.jpg
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micro_pix
Posts: 455
Joined: Fri May 11, 2012 12:05 pm
Location: Southampton, Hampshire, UK

Re: Algae galore

Post by micro_pix »

Very nice! I especially like the last two. Adding a little Calcofluor White fluorescent stain will light up the spirogyra cell wall under UV but I don’t think the result would have looked as good as your images.

Dave

Sumguy01
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Joined: Mon Jan 28, 2013 11:05 pm
Location: Ketchikan Alaska USA

Re: Algae galore

Post by Sumguy01 »

=D> Very nice.
Thanks for sharing.

SW_Perspective
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Joined: Tue Dec 02, 2014 10:29 am
Location: Albuquerque, NM, USA

Re: Algae galore

Post by SW_Perspective »

The third shot with the vorticella is a nice capture. Nice work.
David Moerman
35N 106W
Olympus BH-2, BHTU
NFK relay lens, Splan PL objectives
Nikon D7000, D7200

ModelZ
Posts: 38
Joined: Sun Nov 01, 2020 4:34 am
Location: Northern Europe

Re: Algae galore

Post by ModelZ »

Thanks micro-pix, Sumguy01 and SW_Perspective. Appreciate your comments.

I haven't yet played with fluorochromes at all. One can do wonderful things with them that's for sure.
Here the fluorescing chloroplast is encased in a tube which I wanted to show, but only in silhouette since otherwise the emission from the whole shell would mix up the chlorophyl emission, too. I think here treating the shell with a fluorochrome would not result in as clean image as illuminating the "equator" of the cross section only.

Cheers, Kari

Fortesmentum
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Joined: Fri Feb 26, 2021 11:33 am
Location: Lancashire, England

Re: Algae galore

Post by Fortesmentum »

This is a fascinating thread with good quality photographs ModelZ

I am interested in how it is you utilize UV light in your last two photographs. If possible, would you be able to share a little bit about your set up please?

Thanks

ModelZ
Posts: 38
Joined: Sun Nov 01, 2020 4:34 am
Location: Northern Europe

Re: Algae galore

Post by ModelZ »

Thanks Fortesmentum for comments & interest.

The UV used here comes through Zetopan Epi-fluorescence head. I had to work a bit on it & the light source to use LEDs (Nichia 365 here) and the filter revolver to extend the palette (vanilla ex/em UG1/LP430 here). This brings about the chlorophyl fluorescence, the helix. Spirogyra shell does not fluoresce under this excitation, not in a way that it could compete with the strong red anyway. So the shell is lighted with DF green (from another illuminator). Actually one wants just the shell cross section "equator" lit - this is a bit tricky but with some practice doable. Shooting stacks is essential here. 40x Lumin in both.
If several Spirogyra are in view, ideally you'd like them lying in the same plane i.e. come simultaneously to focus & to right DF light. Of course this never happens.... but you can get close sometimes. Other problems are the helices contracting or moving about during the stack and eventually fading under the UV. You want to do all your prep. under weak green etc. and only turn UV on when all is set.

Fortesmentum
Posts: 2
Joined: Fri Feb 26, 2021 11:33 am
Location: Lancashire, England

Re: Algae galore

Post by Fortesmentum »

Thanks ModelZ for your comprehensive reply.

It sounds like quite a complex set up, but the results are well worth it. The last two photographs of the spirogyra are remarkably good - quite sculptural in appearance.

Smokedaddy
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Joined: Sat Oct 07, 2006 10:16 am
Location: Phoenix, Arizona
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Re: Algae galore

Post by Smokedaddy »

ModelZ,

Mind posting some pictures of your Zetopan setup here, or send me a few pictures?

-JW:

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