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diatom fluorescence comparison

 
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mtuell



Joined: 11 May 2016
Posts: 268
Location: Tucson, AZ

PostPosted: Sat Feb 17, 2018 3:40 pm    Post subject: diatom fluorescence comparison Reply with quote

Long time no see...

Here we have a 40x comparison of Phase Contrast (Ph3 fluor) with UV and blue excitation.



And next 40x Hoffman Modulation Contrast with blue and green excitation.


Nikon Diaphot TMD with Nikon D3300 at ISO 200, single (long) cropped exposures. I let Photoshop do some "Auto-Levels", so the colors aren't as seen necessarily.

Mike
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leonardturner



Joined: 14 Mar 2013
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Location: Atlanta, GA, USA

PostPosted: Sun Feb 18, 2018 2:28 pm    Post subject: Reply with quote

Striking images, Mike. Thanks for posting!
Leonard
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mtuell



Joined: 11 May 2016
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Location: Tucson, AZ

PostPosted: Sun Feb 18, 2018 11:13 pm    Post subject: Reply with quote

Thanks!

Here is a larger Hoffman image.



Mike
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Pau
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Joined: 20 Jan 2010
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Location: Valencia, Spain

PostPosted: Mon Feb 19, 2018 2:04 am    Post subject: Reply with quote

Mike, I have not clear what are you doing here, could you clarify?
- are they live or fresh diatoms or stained in a prepared slide? With live diatoms I would expect all cells providing the same kind of fluorescence, mainly red emission of chlorophylles
- How do you provide the fluorescence excitation light? Epi with the typical filter cube, external incident, transmitted....

Anyway interesting images.
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mtuell



Joined: 11 May 2016
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Location: Tucson, AZ

PostPosted: Mon Feb 19, 2018 10:30 am    Post subject: Reply with quote

Pau,

Yes, sorry. They are just a drop of water in a well slide - there are both live and dead diatoms. The live ones fluoresce, as you say.

This is a Diaphot with EF attachment with commercial cubes and a mercury lamp. Nothing odd or home-made.

The top left is phase contrast through the UV cube, so a clear emission filter. The bottom left is PC through the blue filter, with a yellow emission filter.
The right side is the same thing, but with the mercury lamp beam unblocked.

The bottom set is with Hoffman - blue excitation on top and green on the bottom.

Thanks for taking the time to ask for clarification. I hope this helps.

Mike
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Pau
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PostPosted: Mon Feb 19, 2018 10:53 am    Post subject: Reply with quote

Thanks, Mike!
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GaryB



Joined: 29 Jul 2016
Posts: 519

PostPosted: Mon Feb 19, 2018 5:23 pm    Post subject: Reply with quote

Hey Mike, Nice fluorescing!

I can see I'll have to get one of those UV LED's and see if I can't blind myself, erm, see some nice colored pictures Very Happy
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