Acridine orange adventures

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Wes
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Joined: Sat Mar 09, 2019 12:58 pm

Acridine orange adventures

#1 Post by Wes » Thu Dec 26, 2019 3:31 pm

Acridine orange is a fluorescent dye that can be excited with blue-cyan light and upon binding do DNA and RNA it fluoresces in green and red respectively (there are of course other things that make it glow green or red). Here I added the tiniest drop (< 100 nanoliters) of an acridine orange stock solution to a drop of pond water and captured a few images I decided to share. All images were acquired with the following fluorescent setup: 450-490 bandpass excitation filter, 510 dichroic mirror and 515 longpass filter.

Image
A paramecium I think. Plan 16/0.32

Image
Merismopedia. Neofluar 40/0.75

Image
Colonial algae. Neofluar 40/0.75

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75RR
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Re: Acridine orange adventures

#2 Post by 75RR » Thu Dec 26, 2019 4:49 pm

wonderful images ... well done.

I see the upgrade you mentioned is paying dividends!

viewtopic.php?f=24&p=73559#p73559
Zeiss Standard WL (somewhat fashion challenged) & Wild M8
Olympus E-P2 (Micro Four Thirds Camera)

Wes
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Re: Acridine orange adventures

#3 Post by Wes » Thu Dec 26, 2019 7:34 pm

Thanks 75RR. Its very refreshing to learn new techniques.

PeteM
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Re: Acridine orange adventures

#4 Post by PeteM » Thu Dec 26, 2019 9:39 pm

Very cool images.

And seems even micro-holiday spirited: deck the slides with drops of acridine, fa la la la la la . . .?

Wes
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Re: Acridine orange adventures

#5 Post by Wes » Thu Dec 26, 2019 10:40 pm

Thanks Pete. The green and red touch certainly adds to the Christmas spirit.

charlie g
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Re: Acridine orange adventures

#6 Post by charlie g » Fri Dec 27, 2019 8:54 pm

Thanks for these beautiful results, Wes. Was that first image a ciliate, and was that protozoan still active ( as in a gentle vital strain treatment with Janus Green)?

It would be good to know how long you exposed these pond organisms to your dilute stain ( approx. that is), before these stain effects manifested. Thanks for sharing this bench work, Wes.

Charlie guevara

Wes
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Re: Acridine orange adventures

#7 Post by Wes » Sat Dec 28, 2019 8:57 am

charlie g wrote:
Fri Dec 27, 2019 8:54 pm
Thanks Charlie

The majority of ciliates I spotted were moving around actively. The samples were exposed to AO from anywhere between 10 to 40 minutes. I think the intense illumination is what kills or at least impairs them after a period of observation. A rather dramatic end meets amoeba species stained with AO. They possess intensely fluorescing lysosomes (stained orange due to the low pH) and after about a minute or less of observation the lysosomes suddenly dissolve and the cell disintegrates in the matter of seconds. Below is an example before it disappeared.

Image

billbillt
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Re: Acridine orange adventures

#8 Post by billbillt » Sat Jan 04, 2020 6:32 pm

BEAUTIFUL IMAGES!...

THE BEST,
BILLT

tgss
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Re: Acridine orange adventures

#9 Post by tgss » Sat Jan 04, 2020 9:01 pm

Awesome shots Wes!
tgss

Wes
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Re: Acridine orange adventures

#10 Post by Wes » Thu Jan 09, 2020 5:27 pm

Thank you billt and tgss!

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