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testing 123... (fluorescence and stepper)
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Charles Krebs



Joined: 01 Aug 2006
Posts: 5761
Location: Issaquah, WA USA

PostPosted: Sat Jan 28, 2017 1:53 am    Post subject: testing 123... (fluorescence and stepper) Reply with quote

I've been playing around with a few "new" things. I motorized the MM-11 for stacking and also finally tried a little fluorescence with it as well. These are my first "test" images with the new additions.

The fluorescence is done with a Nichia 365nm LED and a Thorlabs 420nm long-pass filter (#FGL420, $25 for 1" circular). I used the Nichia LED directly and did not add anything to trim any potential minor "spill" above 410nm. I need to test a little more to see if this occurs. If so I will add a sharp bandpass filter to it... but it appears at first look to be doing a decent job in this regard.

The stepper arrangement uses a 0.9 degree stepper motor (400 steps per revolution from WeMacro) and the WeMacro controller and Android control program. It drives the Nikon fine focus knob (200 microns per revolution) directly and thus provides 1 micron movements for each full motor step. (I can go much smaller with micro-stepping but 1 micron will be fine for the objectives I generally use with this microscope).

This first image is a bright green moss


This was a leaf collected at the edge of a marsh. It was bright "kelly" green all over, and I have no idea what I'm seeing with the fluorescence! Smile 10X


Same plant, different leaf with 20X


On my workbench I found a jewel beetle I had previously photographed. The eye fluoresced a very bright "baby" blue. But everything else was black so I added a little diffused "white" light which created an odd looking beetle indeed!

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Last edited by Charles Krebs on Sat Jan 28, 2017 11:23 am; edited 1 time in total
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Jacek



Joined: 02 Oct 2011
Posts: 4689
Location: Poland

PostPosted: Sat Jan 28, 2017 2:48 am    Post subject: Reply with quote

You're the best, you are the champion Very Happy

You did not use excitation filter?
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harisA



Joined: 03 Jul 2011
Posts: 433
Location: Greece

PostPosted: Sat Jan 28, 2017 3:05 am    Post subject: Reply with quote

Excellent images Charles.Will you show us an image of your setup?Is this epi illumination or you illuminate the sample using an external led light source and a diffuser?
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Wim van Egmond



Joined: 15 Aug 2006
Posts: 825
Location: Berkel en Rodenrijs, the Netherlands

PostPosted: Sat Jan 28, 2017 4:39 am    Post subject: Reply with quote

Super results, Charlie. This will be a productive year, I am sure!
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ChrisR
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Joined: 14 Mar 2009
Posts: 7183
Location: Near London, UK

PostPosted: Sat Jan 28, 2017 7:32 am    Post subject: Reply with quote

Well done Charles - the start of another little journey Very Happy
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Cactusdave



Joined: 09 Jun 2009
Posts: 1523
Location: Bromley, Kent, UK

PostPosted: Sat Jan 28, 2017 8:18 am    Post subject: Reply with quote

Very nice indeed. Inspiring. I think the red fluorescing cells on the back of the leaf are likely a unicellular terrestrial alga, a bit like Pleurococcus. A regular brightfield image would confirm.
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fpelectronica



Joined: 19 Feb 2010
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Location: España

PostPosted: Sat Jan 28, 2017 8:41 am    Post subject: Reply with quote


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JH



Joined: 09 Mar 2013
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Location: Vallentuna, Stockholm, Sweden

PostPosted: Sat Jan 28, 2017 9:23 am    Post subject: Reply with quote

Shocked OK so this is testing...
Regards Jörgen Hellberg
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Beatsy



Joined: 05 Jul 2013
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Location: Malvern, UK

PostPosted: Sat Jan 28, 2017 11:24 am    Post subject: Reply with quote

Smashing images - as always.

Could you briefly describe how you attached the wemacro motor to the microscope's fine focus control?

Thanks
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Charles Krebs



Joined: 01 Aug 2006
Posts: 5761
Location: Issaquah, WA USA

PostPosted: Sat Jan 28, 2017 12:09 pm    Post subject: Reply with quote

Thanks all for the kind remarks...

(Note: I corrected the Thorlabs part number for the longpass filter in the initial post. It is an FGL420. (See https://www.thorlabs.com/newgrouppage9.cfm?objectgroup_id=999&pn )

Jacek
Quote:

You did not use excitation filter?


No, not yet. I do have a filter from Ebay that cuts very hard anything above 390nm but I have not tried it yet. I also have a 435nm long-pass filter that could be used instead of the 420nm, but once again I have not tried it yet.

The spectral curve for the Nichia LED looks pretty good, but I might be cutting things a bit too close with the 420nm long=pass and the "naked" LED. (Opinions welcome... this is all new to me. What would I expect to see if the tail of the LED spectral curve edged slightly into the long-pass filter range?)



Beatsy
Quote:
Could you briefly describe how you attached the wemacro motor to the microscope's fine focus control?
Better done with a photo or two...
I'll take a few shots and put together a post over in the "Equipment" or "Techniques" section.

harisA
Quote:
Charles.Will you show us an image of your setup?Is this epi illumination or you illuminate the sample using an external led light source and a diffuser?
I'll cover this in a more detailed post. When I put it up I'll link to it from here.
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Pau
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Joined: 20 Jan 2010
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Location: Valencia, Spain

PostPosted: Sat Jan 28, 2017 12:28 pm    Post subject: Reply with quote

Excellent tests results...and pictures of course.

Could you also post your fluorescence illumination setup?

About the LED, the graph suggests a good "monochromatic" character. Do you see any color (violet...) in its emission? (of course don't look directly at it) . Please link the Nichia model you're using.

Omega, one of the main makers, has a surplus ebay store with good interference filters at much reduced price http://www.ebay.com/sch/m.html?item=310793801058&_ssn=bjomejag&_sop=10
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Charles Krebs



Joined: 01 Aug 2006
Posts: 5761
Location: Issaquah, WA USA

PostPosted: Sat Jan 28, 2017 9:21 pm    Post subject: Reply with quote

I posted two topics to answer a few questions.....

Fluorescence arrangement:
http://www.photomacrography.net/forum/viewtopic.php?p=204217#204217

Stepper motor on MM-11:
http://www.photomacrography.net/forum/viewtopic.php?t=33121
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WalterD



Joined: 06 Jul 2015
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PostPosted: Sun Jan 29, 2017 4:49 am    Post subject: Reply with quote

Lovely set, a pleasure to watch.
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Sumguy01



Joined: 28 Jan 2013
Posts: 883
Location: Ketchikan Alaska USA

PostPosted: Sun Jan 29, 2017 4:06 pm    Post subject: Reply with quote

Very Happy Very nice.
Thanks for sharing.
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René



Joined: 08 Oct 2006
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PostPosted: Mon Jan 30, 2017 2:34 am    Post subject: Reply with quote

Wow! Looking forward to further postings this year!

René
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