diatom fluorescence comparison

Images made through a microscope. All subject types.

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mtuell
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diatom fluorescence comparison

Post by mtuell »

Long time no see...

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

Image

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

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

leonardturner
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Post by leonardturner »

Striking images, Mike. Thanks for posting!
Leonard

mtuell
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Post by mtuell »

Thanks!

Here is a larger Hoffman image.

Image

Mike

Pau
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Post by Pau »

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.
Pau

mtuell
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Post by mtuell »

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

Pau
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Post by Pau »

Thanks, Mike!
Pau

GaryB
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Post by GaryB »

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 :D

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