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Closterium, auto-fluorescence, stereo (red overload!)

 
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Charles Krebs



Joined: 01 Aug 2006
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Location: Issaquah, WA USA

PostPosted: Mon Mar 20, 2017 9:30 pm    Post subject: Closterium, auto-fluorescence, stereo (red overload!) Reply with quote







These last three are not stereo. In the last image, lacy "tree-like" sections initially fluoresced much more than seen here, but they faded fast.






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Jacek



Joined: 02 Oct 2011
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Location: Poland

PostPosted: Tue Mar 21, 2017 12:44 am    Post subject: Reply with quote

Super !
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GaryB



Joined: 29 Jul 2016
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PostPosted: Tue Mar 21, 2017 2:05 pm    Post subject: Reply with quote

That is crazy good.

I have no experience of fluorescence. Do you use a top light, fluor cube/dichoic filter and epi lens etc?

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Charles Krebs



Joined: 01 Aug 2006
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Location: Issaquah, WA USA

PostPosted: Tue Mar 21, 2017 7:21 pm    Post subject: Reply with quote

GaryB wrote:
I have no experience of fluorescence. Do you use a top light, fluor cube/dichoic filter and epi lens etc? Question


Look through this thread to see how I set up:
http://www.photomacrography.net/forum/viewtopic.php?t=33123

In this case I am using external lights instead of an epi-illuminator. I use two different excitation wavelengths (~365nm UV and now ~455nm "Royal Blue") to illuminate the subject on the stage. The microscope has a "barrier filter" inserted that blocks the wavelengths emitted by the lights and only allows the longer wavelengths of the subject fluorescence to pass through. In basic principle, similar to the compound microscope fluorescence illuminators that use epi-illumination and various filter cubes. Chlorophyll auto-fluoresces as deep red under both of my light types.
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GaryB



Joined: 29 Jul 2016
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PostPosted: Tue Mar 21, 2017 10:21 pm    Post subject: Reply with quote

Thanks, Charles.

I love these DIY solutions, especially when the results are so good.
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pwnell



Joined: 18 Dec 2009
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Location: Tsawwassen, Canada

PostPosted: Wed Mar 22, 2017 12:07 pm    Post subject: Reply with quote

Magnificent.
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carlos.uruguay



Joined: 23 Feb 2012
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PostPosted: Wed Mar 22, 2017 1:22 pm    Post subject: Reply with quote

Very nice as usual
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fpelectronica



Joined: 19 Feb 2010
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Location: EspaƱa

PostPosted: Thu Mar 23, 2017 12:14 pm    Post subject: Reply with quote

Very nice
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curt0909



Joined: 26 Oct 2011
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PostPosted: Thu Mar 23, 2017 1:42 pm    Post subject: Reply with quote

Those stereo images are amazing! Great work
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WalterD



Joined: 06 Jul 2015
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Location: Rotterdam, the Netherlands

PostPosted: Thu Mar 23, 2017 10:25 pm    Post subject: Reply with quote

Nice catch!
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ChrisR
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Joined: 14 Mar 2009
Posts: 6843
Location: Near London, UK

PostPosted: Fri Mar 24, 2017 2:42 am    Post subject: Reply with quote

Charles, have you seen significant difference, between the results from the two different excitation frequencies?
There may be a shift in the emission frequency, but if it's still overwhelmingly "red" to a sensor, then perhaps it would be indistinguishable.

Edit - I notice this was largely answered in a different thread. Easier to filter out the UV than the blue??
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Gerd



Joined: 05 Aug 2006
Posts: 152
Location: Duesseldorf, Germany

PostPosted: Fri Mar 24, 2017 12:52 pm    Post subject: Reply with quote

Hi Charly,
woaah, the stereos are absolutely stunning results with your setup, overwhelming indeed. With my filter-cube epi illumination there is a terrible blur all over the details, your images look like confocal images ! With your setup, do you have a visible red blur ? And another question: I guess, these are stacks ? Your image-processing is really perfect !
Kind regards,
Gerd
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