Fluorescent setae of oligochaeta and dead water fleas

Images made through a microscope. All subject types.

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Starshade
Posts: 133
Joined: Wed May 04, 2011 1:47 am

Fluorescent setae of oligochaeta and dead water fleas

Post by Starshade »

Image
Image
Image

Decided to dredge up some my old pictures and found some shots of fluorescent garbage.

The line visible in the shots of the oligochaeta represent the edge of the coverslip

You can check out my post to see some more pics
http://notes-from-dreamworlds.blogspot. ... -junk.html

dariuskersulis
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Location: Boise, ID

Post by dariuskersulis »

Where do you buy your fluorescent dyes from? I'm totally new to this but want to try. Where would you recommend to start? How do you do it?

Starshade
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Post by Starshade »

I work in a lab where everything is bought by lab managers... but you can get these dyes anyway from various companies. Just notice most of them are toxic, carcinogens, or can burn your lungs if you inhale lots of vapor. Also you will need pipettes for dilution, gloves, draft hood, and some basic understanding of what they do and possibly loads of money for the fanciest dyes. Acridine orange is a one of the least poisonous and cheap.

dariuskersulis
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Location: Boise, ID

Post by dariuskersulis »

Okay, sounds like I shouldn't do it. I've read some more of your posts and seems like it's ain't easy either with lot's of trial and error. Your images are beautiful. And the video you've made is just spectacular. Too bad it's more effort and time than you're willing to give to do another one. Your video and photographs from several other guys, to name a few, Charles Krebs and Wim was one of the great inspirations that led to buying a microscope. I'm totally hooked now. Please find some time to create more of your photomicrograps, they have quite unique style.

Starshade
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Post by Starshade »

Thank you. I am absolutely fine with amount of time I spend on pics and video... I love tricky things! The problem is - I need to get this time from somewhere. A good solution would be buying my own scope, cool recording gear and cameras, move to a remote Pacific island, get a boat and stuff, and keep doing all this microscopy as much as I want. I love that idea! Just I am missing some tiny sum of money to accomplish it=) Working on it... I don't give up easily.

Mitch640
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Post by Mitch640 »

What's #3? Looks like a nuclear frog. :)

dariuskersulis
Posts: 54
Joined: Thu Feb 23, 2012 6:38 pm
Location: Boise, ID

Post by dariuskersulis »

haha, good for you! Time is finite resource, I'm also juggling between work, family and the hobby, besides stuff is expensive. The lucky thing for me is that I already live on the island so I'm looking forward to lots of personal discoveries. I wish you to get your money together sooner than later, keep it up!

By the way very cool view rotation effect in the video. Did you simply rotate the phototube while recording? I'm quite comfortable with video editing software and photoshop. Hit me up, perhaps I could take some of the workload in creating one of your video projects. I believe I have huge diversity where I live and equipment up for the job. With some practice perhaps you could even use some of my footage. Anyways, as I said I thought your video was very inspiring, and I'm sure a lot of people also thought the same, and I could use some of my resources to help you get another production. let me know.

Starshade
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Joined: Wed May 04, 2011 1:47 am

Post by Starshade »

Mitch: #3 is a baby of polyphemus pediculus. It's was a rotting embryo that I found in a muddy pond, it had fungi on top and ciliates inside. It looked so disgusting that I had to use some dyes to fix it=)

Darius: Thanks a lot. Biodiversity is a good thing, because where I made the movie it does not exist... you know that if you read the story about making the movie. Marine life is what I miss so much.

In the same post I had a picture of the scope, if you look closely on it you will see that it has a rotating table as it's a scope designed for polarized light microscopy. I hated this table because it is so uncomfortable and does not have control screws, but it was awesome when it came to rotating effects. But you are right - it can be easier to rotate the camera unless the table has some angle... just sometimes it requires extra pair of hands.

At this point I am trying to attract attention of "big guys" by means of participating in movie festivals, because a proper movie needs budget that is beyond capabilities of a mortal researcher. It's gear that's limiting... to make it perfect I would need many special sets to move the field of view smoothly for example. Not to mention a real camera.

But it's good to know that there are motivated people.

dariuskersulis
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Location: Boise, ID

Post by dariuskersulis »

I did read the story, that's why I kind of brought it up. It's funny you mentioned bigger production on microworld. I also though "Planet Earth" is missing an episode.

This is also what I was trying to convey to Zeiss sales rep. (was going to buy Zeiss Primostar before I came upon used Nikon with DIC setup) the difference between astrophotography and photomicrography. At the core, telescopes and microscopes are just the optics, the subject arguably is equally fascinating (while microscope has way bigger selection of subjects), the first one has so many TV shows, documentaries and magazines. Not saying solely because of that, but certainly one of the reasons it being way more popular. While I was barely able to find one video podcast on microbes. The "cool" factor is just not there for general public to get into it.

There is definitely a market for that, the question is if it's large enough to justify the cost so at the end it financially pays off. Perhaps that's why we haven't seen big production on microworld. I'm most likely missing something but I thought your 10 minute video had the best in production value on the subject out there.

curt0909
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Post by curt0909 »

Beautiful photos.

Starshade
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Post by Starshade »

Well, Darius,

just a piece of good news: my movie got a nomination in a wildlife film festival and it will be shown in front of top bbc and AnPl filmmakers... So, there's a chance.

And another thing that came up recently is invitation as a keynote speaker to a large conference on science and art (to talk about photomicrography), can't figure why me , but good possibility of promoting amateur microscopy again.

discomorphella
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Post by discomorphella »

I would be very wary of using most of the fluorescent dyes in a home setting. None of the acridine orange family is really safe to use without gloves and proper precautions. You don't want to track mutagenic chemicals all over your house. And anyone who works with these dyes in a lab will tell you, that if you try measuring out say Fluorescein, you'll find it ALL over the lab in little spots later, unless you double-glove, and immediately discard the outer pair of gloves. The fluorescein dyes and their relatives used in bright-field (e.g. eosin Y, phloxine B and other fluorones) are fairly safe to use protein stains. So is Calcofluor, it specifically stains cellulose and chitin. If you are lucky enough to get a sample, or want to purchase them, you can find fluorescently labeled antibodies to various cellular components too (they will not be cheap, and you will have to know the correct protocols to employ them). You should preferably confine your use to an absorbent-towel-covered workbench so you don't track them all over and of course wear gloves and safety glasses. I can't stress that enough.

Here are some examples of epiflorescence microscopy using Eosin Y and Ploxine B in just regular histological sections.

http://photomacrography.net/forum/viewt ... highlight=

http://photomacrography.net/forum/viewt ... highlight=

Although there are very useful fluorochromes out there for nucleic acid staining and 100's of other histochemical fluorescent methods, its not a good idea to use them outside of a laboratory setting unless you have the training and experience to use them safely.
That being said, you can get plenty of good pictures using natural autofluorescence and simple fluorone dyes.


The image of the rotting leaf in the blog is superb. What dyes did you use?
David

dariuskersulis
Posts: 54
Joined: Thu Feb 23, 2012 6:38 pm
Location: Boise, ID

Post by dariuskersulis »

Haha, not surprised at all. Good for you! Keep us posted on what's happening. I think I liked your Facebook page but rarely anything comes through, keep it up ;)

Thanks David for the scare on flourescent dyes, I have two kids so definately don't wan't to keep anything more hazardous than cleaning supplies.

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