testing 123... (fluorescence and stepper)

Images made through a microscope. All subject types.

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

Post by Charles Krebs »

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
Image

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! :) 10X
Image

Same plant, different leaf with 20X
Image

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!
Image
Last edited by Charles Krebs on Sat Jan 28, 2017 11:23 am, edited 1 time in total.

Jacek
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Post by Jacek »

You're the best, you are the champion :D

You did not use excitation filter?

harisA
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Post by harisA »

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?

Wim van Egmond
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Post by Wim van Egmond »

Super results, Charlie. This will be a productive year, I am sure!

ChrisR
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Post by ChrisR »

Well done Charles - the start of another little journey :D
Chris R

Cactusdave
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Post by Cactusdave »

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.
Leitz Ortholux 1, Zeiss standard, Nikon Diaphot inverted, Canon photographic gear


JH
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Post by JH »

:shock: OK so this is testing...
Regards Jörgen Hellberg
Jörgen Hellberg, my webbsite www.hellberg.photo

Beatsy
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Post by Beatsy »

Smashing images - as always.

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

Thanks

Charles Krebs
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Post by Charles Krebs »

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. ... _id=999&pn )

Jacek
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?)

Image

Beatsy
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
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.

Pau
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Post by Pau »

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=310 ... ag&_sop=10
Pau

Charles Krebs
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Post by Charles Krebs »

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

Fluorescence arrangement:
http://www.photomacrography.net/forum/v ... 217#204217

Stepper motor on MM-11:
http://www.photomacrography.net/forum/v ... hp?t=33121

WalterD
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Post by WalterD »

Lovely set, a pleasure to watch.

Sumguy01
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Post by Sumguy01 »

:D Very nice.
Thanks for sharing.

René
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Post by René »

Wow! Looking forward to further postings this year!

René

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