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Autofluorescence in mosses.

 
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WalterD



Joined: 06 Jul 2015
Posts: 412
Location: Rotterdam, the Netherlands

PostPosted: Wed Nov 21, 2018 11:47 am    Post subject: Autofluorescence in mosses. Reply with quote

After the sporangia I wanted to jump back to mosses to observe autofluorescence. (Determination for follow up!)
The first 2 pictures are showing a moss with leafs that cannot be seen with the bare eye; looks like green velvet. Picture 1 shows a side view, 2 a top view which is more difficult to stack. The sample was found in an industrial environment.

Picture 3 shows the spore capsules of a moss with very small hair-like leafs. Noted mosses can have spore capsules althrough the year in Europe.

When taking picture 4 the photobleaching was (as always) an issue. I observed the bleaching started in the tips of the leafs and propagated to the inside. Meantime the leafs started to curl up. Within the first 20 seconds of exposure the object lost about 75% of its radiation. The same method was used as in the previous posts last 2 pictures: obscuring the mercury vapour lamp any time the camera sensor was not exposed.

Picture 5 is a better illustration of the "glow in the dark" effect of autofluorescence on a moss leaf, generated by the chlorophyl under UV excitation. The tip of that leaf is asking for a higher magnification...
1.


2.


3.


4.


5.


All done with UV excitation, YOC-3 filter and 9x epi-objective (3,4,5), 7x (1,2).
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Jacek



Joined: 02 Oct 2011
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Location: Poland

PostPosted: Wed Nov 21, 2018 12:31 pm    Post subject: Reply with quote

Very nice
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Beatsy



Joined: 05 Jul 2013
Posts: 1431
Location: Malvern, UK

PostPosted: Wed Nov 21, 2018 12:43 pm    Post subject: Reply with quote

Interesting set. Is there such a thing as UV flash so as to reduce the photo-bleaching issue?
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Sumguy01



Joined: 28 Jan 2013
Posts: 1196
Location: Ketchikan Alaska USA

PostPosted: Wed Nov 21, 2018 4:52 pm    Post subject: Reply with quote

Very nice.
Thanks for sharing.
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Marek Mis



Joined: 10 Jul 2010
Posts: 2522
Location: Suwalki, Poland

PostPosted: Thu Nov 22, 2018 12:45 am    Post subject: Reply with quote

Very nice images Walter, especially the first one !

Marek
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WalterD



Joined: 06 Jul 2015
Posts: 412
Location: Rotterdam, the Netherlands

PostPosted: Thu Nov 22, 2018 1:12 pm    Post subject: Reply with quote

My pleasure




Quote:
(Beatsy) Interesting set. Is there such a thing as UV flash so as to reduce the photo-bleaching issue?

I wish there was. The mercury arc vapour lamp needs to burn at least 2 hours each session, otherwise it will not last the 200 burning hours. There are diaphragm shutters with controllers, (https://www.thorlabs.com/newgrouppage9.cfm?objectgroup_id=6619) such could be integrated in the instrument between the lamp and the optics. However, besides introducing another source of vibration, it should be talking with Canon EOS Utility, which is impossible as far as I know.
Another option would be utilizing "smart glass", this is transparant material that darkens when a current is sent through it. Here the same issue: how to trigger anything that's not incorporated in the Canon software? If anybody has ideas please share it here...

Regarding the picture: sorry no ID yet, 7x/0.17 epi objective.
Zerene Dmap substacks.
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rjlittlefield
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Joined: 01 Aug 2006
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Location: Richland, Washington State, USA

PostPosted: Thu Nov 22, 2018 1:19 pm    Post subject: Reply with quote

WalterD wrote:
Canon EOS Utility
...
Here the same issue: how to trigger anything that's not incorporated in the Canon software? If anybody has ideas please share it here...

Consider using a keyboard/mouse "robot" to drive both Canon EOS Utility and the controller(s) for other software. Something like http://www.robot-soft.com/ .

I have worked with several people who use this approach with camera control software to acquire focus stacks.

--Rik
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carlos.uruguay



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

PostPosted: Thu Nov 22, 2018 4:18 pm    Post subject: Reply with quote

Super!!
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WalterD



Joined: 06 Jul 2015
Posts: 412
Location: Rotterdam, the Netherlands

PostPosted: Fri Nov 23, 2018 11:15 am    Post subject: Reply with quote

Thanks Carlos!

Quote:
Consider using a keyboard/mouse "robot" to drive both Canon EOS Utility and the controller(s) for other software. Something like http://www.robot-soft.com/ .

I have worked with several people who use this approach with camera control software to acquire focus stacks.


That's brilliant Rik! What you mentioned is an application mimicking clicking a mouse on a pre-defined location on the screen. This will enable me to use the mouse for the shutter controller simultaneously. With this knowledge it will be a matter of time before it's incorporated within my setup. Anyhow, there's room enough to install it Very Happy
Hopefully I can beat the price barrier, what I spotted sells for about $1000...

This whole issue would be worth a separate post in the equipment discussions section, maybe I'll do that this weekend. Meantime I've created a 6-seconds animation showing the effect of photobleaching and UV light on moss as shown in picture 4 (now under postprocessing/ being retouched) https://www.youtube.com/watch?v=qrFOkjVc5zg&feature=youtu.be
Be warned the compression resulted in poor image quality, the clip is also too dark.
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