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Lichen in a different light

 
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WalterD



Joined: 06 Jul 2015
Posts: 361
Location: Rotterdam, the Netherlands

PostPosted: Sat Mar 03, 2018 11:07 pm    Post subject: Lichen in a different light Reply with quote

"Lichens comprise a fungus living in a symbiotic relationship with an alga or cyanobacterium (or both)"

The other day, while doing a little walk I found a loose branch with moldy bark in the middle of the street. The bark was completely covered with algae, lichens and moss. Some of the lichens present can be seen in below photographs. The first "session" was done with a low magnification objective (Leitz 4x/0.12) with help of 2 of the well-known Ikea reading lamps. A third -red- led was added to the setup in order to get a more 3-dimensional result/ a better look at different details. This explains the pink-lilac shadows in the first 3 pictures. In photo 2 a part of the inside of the lichen can be seen on 2 spots (e.g. lower right), as it was slightly damaged. This inside is also showing a bit of the fungus-like structure. Inside at higher magnification also chlorophyl can be found.


The next three photographs were made with the fluorescent setup and 9x objective. The russian emission filter has not got a narrow bandwith (also called a longpass filter), therefore various wavelengths of radiation are visible. See also my post in equipment discussions.
All in all the first 3 photos are very different from the last 3.

Determination: probably Xanthoria polycarpa (2 & 4) and Physcia tenella (3) and Xanthoria parietina (6). (1) looks somewhat different compared with (2), with deeper and more narrow beakers.
This set can be found @ 1376 pixel width via this link: http://waltermachielsen.com/lichen/







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micro_pix



Joined: 11 May 2012
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PostPosted: Sun Mar 04, 2018 9:20 am    Post subject: Reply with quote

Excellent! Very interesting ways to light/view lichen.
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Lou Jost



Joined: 04 Sep 2015
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Location: Ecuador

PostPosted: Sun Mar 04, 2018 10:06 am    Post subject: Reply with quote

Very beautiful. What wavelength did you use for excitation?
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WalterD



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Location: Rotterdam, the Netherlands

PostPosted: Sun Mar 04, 2018 11:18 am    Post subject: Reply with quote

Glad you like them!
Lou, this is the blue filter block. I translated from the manual (by google) :
Quote
On one plate, a coating is deposited reflecting mainly light in the region from 300 to 380 nm and transmitting light in the region from 420 to 500 nm. This plate is used in the study of blue-blue luminescence, excited by the long-wave region of the ultraviolet spectrum.
Unquote
So 420-500nm, but combined with the ???6-3 exciter filter, which operates in the ultraviolet spectre, see below graph. AFAIK the barrier filter is longpass only blocking UV. Does that make sense? Smile
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ralfwagner



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PostPosted: Sun Mar 04, 2018 11:21 am    Post subject: Reply with quote

Very nice. Really a different and more artistic view.
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Pau
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Joined: 20 Jan 2010
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PostPosted: Sun Mar 04, 2018 11:44 am    Post subject: Reply with quote

Excellent set, I most like the first fluorescence image.

I wouldn't qualify your filter cube as blue, it's a fairy typical UV set.
Quote:
So 420-500nm...

I see red fluorescence so I think this is not the emission filter you're actually using, maybe it's a classical long pass filter.
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Last edited by Pau on Tue Mar 06, 2018 10:50 am; edited 1 time in total
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Lou Jost



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PostPosted: Sun Mar 04, 2018 11:44 am    Post subject: Reply with quote

So excitation was mostly in the UV with the filter blocking visible light, and all the colors we see are fluorescence colors?
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Sumguy01



Joined: 28 Jan 2013
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PostPosted: Sun Mar 04, 2018 12:00 pm    Post subject: Reply with quote

Excellent set.
Thanks for sharing.
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WalterD



Joined: 06 Jul 2015
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Location: Rotterdam, the Netherlands

PostPosted: Sun Mar 04, 2018 12:36 pm    Post subject: Reply with quote

Thanks everybody!
The block I've been using is what the russians call "blue". That word is -in cyrillic- engraved in the block. I just received the missing Ultraviolet block from Ukraine and have compared both utilizing the same barrier filter. Result was I found the Ultraviolet block for this subject the "weaker version" of the blue.
Now the exciter filter (lollipop type) I've been using with the blue block is the YOC6-3. In my previous answer to Lou this was scrambled into question marks as I used cyrillic letters. This YOC6-3 filter has the characteristic as shown in the graph above, which means excitation in the ultraviolet bandwith.
The colorless barrier filter (16mm diameter on top of the block with the dichroic mirror) is expected to be longpass.
So UV exciter filter, blue dichroic mirror and -presumably-longpass barrier filter showing bigger part of the visible spectre, that's what has been used. I must say the choice of lollipop exciter filters is pure trial and error, I'm also not really confident with the translated manual. Rolling Eyes

One more remark about the new UV block: It originally came with a "black" barrier filter blocking all visible light for the major part and only letting through the UV. That one is actually missing, can find it on ebay of course but...does it sound familiar or ring a bell?

Best regards,

Walter
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Pau
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Joined: 20 Jan 2010
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PostPosted: Sun Mar 04, 2018 1:14 pm    Post subject: Reply with quote

Quote:
One more remark about the new UV block: It originally came with a "black" barrier filter blocking all visible light for the major part and only letting through the UV. That one is actually missing, can find it on ebay of course but...does it sound familiar or ring a bell?


The more typical black UV excitation filters are Schott UG1 and UG5 mounted in most old fluorescence microscopes (Zeiss, Leitz, Nikon...)
You can see their spectra at the Schott site https://www.pgo-online.com/intl/schott-filter-calculation/schott-color-glass-filter-calculator.html
Their spectra seem to match Lomo YOC8 and YOC5 aproximately.

I have both UG filters and both work well. I don't use the UG5 because it is fungus affected. The UG5 also works with violet excitation so it could be preferable with fluorophores with excitation spectrum around 400nm

Your excitation filter seems to work very well, so if you can combine it with both beamsplitters I don't see the need of another one.
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WalterD



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PostPosted: Tue Mar 06, 2018 9:40 am    Post subject: Reply with quote

Thank you for the very interesting reply, Pau. Also really appreciated the reference links provided in the "retrofit" post. Below is an overview from Lomo regarding the excitation and transmission bandwiths for each block. As you can see the bandwidths themselves are rather wide, but can be narrowed by specific exciter filters. The 4th column is showing the filters that had to be applied on the eyepiece lens (blocking solely UV) as well, they start with a russian "X". The fifth column indicates the 16mm barrier filter for each block, The fifth block is mirrorless, for epi darkfield/non fluorescence. Now I own all the five blocks!
I discovered another combination of exciter filters with the green filter block, giving below results.
I'm using a full frame DSLR now as well. Furthermore I relocated the collector lens a bit and used a lower magnificatyion epi objective (about 7x magnification).




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Pau
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PostPosted: Tue Mar 06, 2018 10:12 am    Post subject: Reply with quote

Very nice!

Now I understand the labeling: they are named with the emission filter, not with the excitation filter that seems more standard.

So, seeing both your results and the tables you have as I said the standard near UV, blue and and green excitation sets and you don't want to use the short UV one to burn your eyes (at least if you don't have a modified camera for UV that would be interesting)
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Jacek



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PostPosted: Thu Mar 08, 2018 10:15 am    Post subject: Reply with quote

Super Very Happy
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