Closterium, "Royal Blue" LED + animated GIF

Images made through a microscope. All subject types.

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Charles Krebs
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Closterium, "Royal Blue" LED + animated GIF

Post by Charles Krebs »

I put together two "Royal Blue" LED lights and got the appropriate filters. Wanted to try them out for auto-fluorescence, and the best subject I found quickly was closterium. These have been "done" here quite a bit over the past few years and I always found it fascinating to compare the auto-fluorescence image with the regular visible light image. I've started to work up a couple stereo versions and they are really fun to see. I'll post them separately when finished.
50X Olympus LMPLFLN objective. Canon T3i.

Image
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zzffnn
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Post by zzffnn »

Great results, Charles!
So you made the LEDs yourself? Which LED did you use?
Thank you for sharing!
Selling my Canon FD 200mm F/2.8 lens

GaryB
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Post by GaryB »

Ditto on that, it looks amazing and I'd love to know how that works.

Great job.

Charles Krebs
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Post by Charles Krebs »

zzffnn wrote:So you made the LEDs yourself? Which LED did you use?
I used the Cree XT-E. Definitely needed a "clean-up" filter to eliminate some green.

Jacek
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Post by Jacek »

Very nice, super results :D

Peter M. Macdonald
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Post by Peter M. Macdonald »

Charles,

I took the liberty of taking the GIF into Photoshop and making the two frames into layers. The red layer was placed on top, blending mode was set to Luminosity, opacity to 62% and this gives most of the colours of the green version, with much of the internal structure from the red version.

It was then necessary to brighten the image up with a levels adjustment so as to match the luminosity of the original.

Regards,

Peter

Pau
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Post by Pau »

Charles, again perfect pictures!

I find Royal blue clearly better than UV to excite chlorophyll, do you?

Just a caveat line: Usually the royal blue Cree XT-E that is excellent is promoted to work at 1500mA max current drive, likely this is why I burned mine after driving it at 1400mA for few non continuous hours. After taking a more careful look at Cree specifications sheets I found that 1500mA is for the white version while for the blue version it is 1000mA.

Because this difference is not easy to spot at first look, I think they are to blame at 50/50 Cree and the ebay sellers

I'm still waiting the replacement to arrive :(
Pau

Charles Krebs
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Post by Charles Krebs »

Thanks for the kind replies.

Pau... sorry to hear about your Royal Blue LEDs. I'm driving mine at 700mA and using a 10003 Carclo Lens (20mm Narrow Spot LED Optic) in front of it. I was a little worried about the polycabonate lens blocking some desirable light but from what I have seen they pass above 400nm freely. They certainly seem to be at least as effective with chlorophyll as the UV lights.

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Post by Johann »

These are amazing.

Peter, I was thinking exact same line as you, would love to see the image if Charles don't mind you sharing it?

Charles Krebs
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Post by Charles Krebs »

If Peter stops back here, it is fine with me to post the layered version he spoke about. I have been pondering other ways of doing such a comparison myself.

carlos.uruguay
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Post by carlos.uruguay »

Nice!

Peter M. Macdonald
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Post by Peter M. Macdonald »

Charles,

Happy to post. As I never save the results of playing with other members' forum images in Photoshop I have just redone the composite.

If doing this for yourself, it is worth playing with the opacity slider on the "red" layer, as relatively small adjustments can make quite a large difference to the overall look of the result. I liked the look at 62%, others may have chosen a different opacity.

Image

Best wishes,

Peter

curt0909
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Post by curt0909 »

Super comparison. Your DIY fluorescence approach is certainly yielding some excellent results!

Johann
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Post by Johann »

Nicely done Peter - I think that is a great final result - thanks for sharing :)

Peter M. Macdonald
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Post by Peter M. Macdonald »

My pleasure, Johann. I only did the easy bit. Charlie did the difficult bit.

Peter

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