Fluorescent images of Protists

Images made through a microscope. All subject types.

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Robert Berdan
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Fluorescent images of Protists

Post by Robert Berdan »

I am using a Nikon Optiphot refurbished microscope for the pictures below. It is a Wide field Fluorescent microscope with UV and green excitation and I have a Nikon DSLR attached. I recently purchased a 10X and 20X Fluor objective and I am hoping to buy a 40X Fluor objective with numerical aperture > 0.85 in the future. While waiting for the ice to clear off the ponds in Calgary I have been staining microorganisms with acridine orange. The specimens are from my freshwater aquarium - images shown below.

I am working to try and improve images clarity and sharpness - I may have to try some deconvolution software. I am using Photoshop RAW filter to remove haze, glow etc. Acridine orange stains DNA-nuclei green or yellow, RNA and acidic compartments like lysosomes red-orange. Many of the organisms survive for more then 6 hours in Acridine orange in low concentration (1 microgram\ml).

Specimens taken with 10 & 20X Nikon Fluor objectives:
1) Amoeba 2) Peritrich with yellow macronucleus and no stalk - Vorticella sp? 3) Bdelloid rotifer 4) Gastrotrich 4) Amoeba 5) Halteria grandinella - fast moving ciliate. Exposures are between 1-5 seconds ISO 400 - fast moving organisms are often blurred under UV light due to movement and realtively long exposures .

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RB

micro_pix
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Post by micro_pix »

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Very impressive results!

Charles Krebs
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Post by Charles Krebs »

Nice, and an abituous project! The only live "critter" I've illuminated with UV was a Hydra viridis that was on a slide I was working. It fluoresced bright red but completely recoiled at the UV light.

Robert Berdan
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Joined: Wed Oct 18, 2017 8:58 pm
Location: Calgary
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Thank you for your comments

Post by Robert Berdan »

If you are interested - I just posted the full article and compare different Deconvoluton procedures I tested to increase the image clarity.

https://www.canadiannaturephotographer. ... scopy.html

Thank you for you kind comments
RB

micro_pix
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Joined: Fri May 11, 2012 12:05 pm
Location: Southampton, Hampshire, UK

Re: Thank you for your comments

Post by micro_pix »

Robert Berdan wrote:If you are interested - I just posted the full article and compare different Deconvoluton procedures I tested to increase the image clarity.

https://www.canadiannaturephotographer. ... scopy.html

Thank you for you kind comments
RB
Thanks for the link Robert - that's a great article, I'm going to have a go at some fluorescence microscopy and you've inspired me. I did have a try at autoflourescence under UV but the subjects are a little limited so I will try and get hold of some Acridine.

David

Smokedaddy
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Post by Smokedaddy »

Excellent

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