Homemade Micro-Aquariums and Slide Ringing Table

[KurtM]

A few months ago I posted a web page discussing my homemade micro-aquariums and slide ringing table, see:
http://sawdustfactory.nfshost.com/Micro%20Aquariums/

The little aquariums have been just the thing for observing wee pond critters in a natural setting using a stereo microscope head on a photo tripod via homemade adapter and macro photography focus rail. I can get lost for hours gazing into this little habitat...



Good for macro photography too, here using a Canon T3i, Canon 60mm macro lens, extension tubes, focus rail, and operating the camera remotely with Canon Utilities software on a laptop computer.




Which gets me pictures like this:


Look Ma, Daphnia thinks the Volvox are beachballs!



A Daphnia chorus line!



Hydra and friends (as long as they steer clear of those tentacles).


See more on my Photobucket album at:
http://s70.photobucket.com/user/Arky704 ... t=6&page=1

And Flickr:
https://www.flickr.com/photos/67904872@ ... 912223623/



On the same page I have posted details about my slide ringing table, read all about it at:
http://sawdustfactory.nfshost.com/Micro%20Aquariums/

[75RR]

Interesting project. Have you tried smaller Micro-Aquariums?
I was wondering if a thinner one, if it were sustainable, would provide for more photographic opportunities.

[KurtM]

First consideration was make 'em easy to maintain so as to have fewest excuses not to use 'em over the long haul. Therefore, they're large enough to get a hand in, for cleaning, because I just happen to know that cleaning a sample container after several months tends to require some serious scrubbing.

Second consideration was to provide just enough volume to get reasonable sustain (sounds like I'm describing an electric guitar).

Mostly I just used the TLAR System to design them. TLAR = That Looks About Right.

PS: If you haven't already, Google micro aquariums, then select Images. Lotsa intriguing eye candy (if you're a hopeless nerd like me I guess).

[zzffnn]

Interesting project. Have you tried smaller Micro-Aquariums?
I was wondering if a thinner one, if it were sustainable, would provide for more photographic opportunities.

I have. Please see my photo at the end of my post. My small and thin culture vessel can be maintained for about 3 months.

I have been using T75 tissue culture flasks (with 75 cm2 surface area) as micro-aquarium. Their polystyrene walls are around 1mm thick (designed for inverted scopes) and optically transparent.

Photographing and lighting them is easy due to their light weight and small size. I used a compound microscope's XY mechanical stage to hold the T75 flask on its slender side and provide XYZ movement (scope's head is chopped off and vertical movement is provided by scope's focusing mechanism) .

My stereo scope (a B&L Srtreozoom 7) is set up simialrly to Kurt's stereo scope, except that it only lays flat on some books or a lab jack - since all required XYZ movements are provided by the beheaded compound scope's stage that moves with the flask - my stereo scope does not need to move. I love how precise the beheaded scope stage can move the flask, as movement has to be precise for magnification of over 40x (my SZ7 goes to 70x).

My set-up is not possible with bigger glass tanks, since compound scope's slide clamp is not strong enough for anything bigger/heavier than T75 flask.

But because of their small volume, T75 flasks are only sustainable for about 3 months. After 3 months, algae die off, the cultures turn from aerobic to anaerobic, then protists die off. All they need is some sunshine and tiny amount of fish food every 3 weeks or so. You may be able to keep the culture longer than 3 months, if you are careful, I did not pay much attention to my cultures at all.

Also smaller sample volume likely means less biodiversity.

Yet another drawback is that such tissue culture flasks are difficult to clean and reuse. But they are cheap to buy from eBay in dozens. Smaller T25 (25cm2) flasks can work the same way too.

The following photo shows you a rough idea. Note that it is a smaller T25 flask without pond culture in the photo. It is too late at night now and I do not want to pull out my real cultures in T75 flasks. Flash or flexible arm holding LED headlight can be easily added for more light.

[rnabholz]

Hey Kurt,

You know that I am a fan of your set up. The shots you are getting are really fascinating. Maybe this winter I will get around to putting together something like it for myself.

Thanks for posting.

Rod

[Crater Eddie]

Great setup and gorgeous shots!
CE

[KurtM]

Thanks, Rod and CE!

And thank you Fan, your comments and photo are very interesting (except there's nothing in your little tank!). First, using the microscope stage to mount your micro-aquarium is brilliant, and guaranteed to work better than my arrangement (while keeping in mind they're two different sets of requirements due to container size/weight). For my rig, getting a tripod and head sufficiently robust to support stereo scope heads and cameras laden with extension tubes, focus rails, etc., is no small thing. It would greatly simplify everything to keep the scope/camera stationary and transfer the motions to the tank. All I need is more time ... (sound familiar?)

Secondly, I like the culture flasks, will order some soon. But I also need better open containers than the plastic and glass 50-60 mm Petri dishes I've gotten so far off eBay. Need better optical quality transparent surfaces for use on my inverted scope. Do you (or anyone else) know of better alternatives?

[zzffnn]

Kurt,

I have attached a photo showing my real pond culture, in a T75 tissue culture flask, laid on the beheaded microscope stand/stage.

I forgot to mention that I only put cap on the flask when I observe it. Otherwise, cap has to be removed to allow oxygenation.

I don't know better quality culture vessel other than petri dish or tissue culture flask. In research labs, those are all they use. There should be ones with better optical quality, but they would probably cost a lot more (many don't demand top optical quality from ordinary inverted scopes - it can be done but will cost a lot more and demands professional set-ups).

Can your inverted AO scope look into petri dish with 40x objective? I know some inverted 40x objectives do not have long enough working distance for petri dish (they can only view cells grown on cover slip mounted invertedly on slide by surface tension). To view petri dish, objectives have to be long working distance 40x corrected for 1-1.5 mm cover, but those are not cheap.

-------------------------------------------------------------------------

To others not familiar with vertical micro aquarium:

The point of this kind of vertical rig is to keep protists in their natural pond layer and let them behave naturally. For example, stentor and many ciliate live at the bottom and hunt like cats. You won't appreciate their way of swimming or other natural behaviors, if you put them on petri dish or slide and view UNDER a scope - you have to view the pond from its side, to see what it is like in nature.

[KurtM]

Fan, I get pretty decent views through the inverted using plastic Petri dishes, but notice some distortion. I purchased some glass Petri's, but so far, more distortion, the plastics are still best. If you run across any in plastic or glass that are made to look *through* I hope you'll alert me (I suspect you know how to shop for lab stuff better than me). Price shouldn't be too big a deal since I don't need many, and they're reusable.

My AO 1810 inverted microscope has three objectives: 6.5X NA 0.2, 10X NA 0.25, 20X NA 0.50, and #1244 LWD condenser on the phase turret. I get dark phase contrast for 10X and 20X only. I once swapped to a 4-place nosepiece and added a 40X dark phase objective and corresponding annulus to the turret but ran into problems and went back to original config. I failed to make notes of what the problems were, and can't recall now, dang it. I think it was along the lines of interference, where the 4-place simply didn't fit; otherwise I'd have most likely left it in...?

[zzffnn]

Kurt,

I heard that glass petri dish is usually a bit thicker than plastic dish. Thicker bottom causes more spherical distortion, especially at objective NA 0.4 and up. That is why inverted objective >= NA 0.4 usually have correction collar for dish bottom ranging from 0.9-1.5mm.

Other than that, culture vessel probably does not matter that much. I have no buying experience with specialized culture vessels.

You may be able to let through more light, by mincing vegetation of your pond culture (vegetation blocks light from condenser and can cause distortion).

I would actually suggest using LWD inverted objectives that are specially designed for using with tissue culture flasks. Unfortunately, your AO 1810 is an infinity scope and can only use infinity objectives (mostly only AO branded ones). You would have to ask Phil or Charles, as I know nothing about those.

I only know about 160TL inverted objectives (which won't work on your AO1810). Those are also not cheap - the cheapest 40x 160TL tissue culture LWD objective that I have seen costs $100 shipped. 20x 160TL versions cost around $50-$80 each. Any regular 10x or lower would work on an inverted scope, since their NA is low and WD is long enough.

[gekko]

I enjoyed looking at the pictures of your setups. I like the ingenuity, simplicity, and elegance. And the images you get are superb! I really had no idea you could get such excellent, detailed pictures (Hydra, Volvox, Daphnia, etc.) using a stereo microscope or a macro camera lens. Great work! And a very instructive post.
Is the large black object above the aquarium in the 2nd picture a flash?

[einman]

My set-ups are not quite as elaborate. Here are 3 "mini aquariums" I use to view live specimens. All three can be used with a stereoscope and two are compatible with a compound scope. The small petri dish is also used to view live insects and small amphibians. The volume of water is not sufficient to sustain larger invertebrates but they can sustain the much smaller species for short periods of time. I have used larger petri dishes and kept them going with ostracods etc. for a month.

[KurtM]

Fan, the distortion in the glass Petri dishes I'm talking about is plainly visible just with a glance at 'em in your hand. The glass was obviously never meant to be looked through at all. I'd like to find some dishes that have bottoms made like the sides of your culture flasks whether glass or plastic.

Gekko, thanks! The precise reason I like posting about this stuff is 'cuz I believe many amateur microscopy enthusiasts who have 'em don't think about using their stereo scopes on pond critters, and are missing out on some great fun. One of the most astounding things you get is a handle on the true sizes of these wee beasties. They're not really microscopic at all, and you even become able to observe them with just the naked eye (and a good pair of glasses ).

This evening I tried out some new toys for the first time: my brand new new tripod-n-head combo, new Canon 100 mm macro lens I got a month or two ago, and lab jack (gettin' fancy). It was just a quickie test drive to get the lay of the land, no flash (batteries were dead, they're charging now lol) or laptop computer remote camera control was used. And yes, the black thing hanging above the tank in the picture way above is a speedlight flash, in my case a Pixel Mago, which is a heck of a bargain as a quick Googling will soon show. It works wirelessly with the Canon T3i camera, very cool!



Which resulted in this snapshot of a trio of very pale hydras on some filthy glass. This micro-aquarium sample was put in late last March, studied through late May, Then totally neglected all summer and fall until just now; but there are still plenty of hydras and ostracods present. I need to more closely examine and see what else. Is this fun or what?



EDIT: It just occurred to me that the micro-aquarium culture in the picture above, and the one here, are actually the very same sample; nothing has been changed, subtracted, or added since last March. It lives on a shelf in a window that does not receive direct sunlight.

[Crater Eddie]

Holy cow! I have never even seen a hydra, and here you have them coming out of your ears.
CE

[zzffnn]

einman,

Please kindly elaborate on your photos #3 and #4, or provide purchase web oage. Photo #4 looks like a quality observation vessel, since walls look thin. I know one can DIY something similar, but I don't have good hands like Kurt.

[zzffnn]

Kurt,

You have a sweet rig there.

Edit: Many glass dishes have uneven bottom, which is another cause of distortion. Plastic ones are easier to make evenly thin. At least that is what I heard.

How do you like that lab jack? Is it stable, sturdy and durable? If so, I would like to buy one too. Please kindly provide purchase web page.

I currently support my SZ7 stereo scope with books, but I would like to use a lab jack instead. I know some budget lab jacks are not very stable, so I have not bought one.

[einman]

Wow I love those hydras!!! That alone is worth the effort!

Fan- I purchased them from Carolina Biological supply. The optics are excellent. The petri dish has nearly 0% distortion and is better than most glass petri dishes for optical clarity.

http://www.carolina.com/protist-viewing ... ?question=
http://www.carolina.com/protist-viewing ... ?question=

[zzffnn]

Thank you einman.
The compression chamber looks especially attractive.

Would you please kindly measure its single wall thickness and reservoir thickness for me? I know its total thickness is 4mm, so I guess single wall thickness should be close to 1mm and reservoir thickness is close to 1mm as well? That dimension is optically ideal, if the whole thing is not fragile.

Kurt, do you think you can DIY something of similar dimension? I don't know if store-bought glass blanks can be very thin, if so, DIY should not be difficult.

[KurtM]

Many glass dishes have uneven bottom, which is another cause of distortion. Plastic ones are easier to make evenly thin. At least that is what I heard.

Well, you've just heard it again, because it exactly describes my experience...

Got the lab jack off eBay. I watched them for a while and tried my best to snag a good one, but still ended up disappointed with cheepie quality. Don't recall what I paid, but it was neither the least nor most expensive - tried to hit somewhere in the middle, feel like I ended up on the low end regardless. But even if it doesn't work well, at least it works. Not sure what you mean by unstable, but I sure wouldn't put my tank on it if I thought it might fail.

CE and einman, two things about hydra: first, chances are you've had them before, and never knew it. Second, if you have one, you have a bunch ... or soon will (given a suitably generous sample volume, that is, say a pint or more). Keep sampling different bodies of water, you never know when you'll run across a rich yield.

[einman]

Thank you einman.
The compression chamber looks especially attractive.

Would you please kindly measure its single wall thickness and reservoir thickness for me? I know its total thickness is 4mm, so I guess single wall thickness should be close to 1mm and reservoir thickness is close to 1mm as well? That dimension is optically ideal, if the whole thing is not fragile.

Kurt, do you think you can DIY something of similar dimension? I don't know if store-bought glass blanks can be very thin, if so, DIY should not be difficult.

Interesting- Carolina's site says 4 mm thick. By my gage the one I have is 5 mm thick. The internal is 1.67 mm. I purchased mine about 2 years ago. It could be they have gotten thinner.

[zzffnn]

Kurt,

I wanted to ask if your lab jack would wobble a little with your tank on it. When viewed at 30x-40x or your highest magnification, do you see image shaking?

I think you tank with water is about the same weight as a stereo scope head. I plan to put my stereo scope on a lab jack. Thank you!

[zzffnn]

einman,

If total thickness is 5mm and your internal space is 1.67 mm thick, then a single wall would be about 1.67 mm thick too, I guess? That might be too thick, as my tissue culture flask is less than 1.5 mm for sure (more likely 1.2mm or less thick). I guess I have to ask Carolina to make sure.

[charlie g]

Hi Kurt and all...fantastic that your hydras have thrived so long with a balance your mini tank affords them...congrats,Kurt!

To this day ( off and on for many decades of freshwater microscopy) I in NY/NJ areas...have not once encountered wild Volvox protists.

It's nice your mini- tank offers that rather stable microcosm. I enjoy only one indoor microcosm which has been hosting duck weed, bryophytes, a tree bark chunk with moss, snails,water fleas,flat worms as it's mioflora/meiofauna for years.

Into it I plop some of the current organisms after veiws with the microscope...guess it's sort of a slop-bucket for organisms after their being observed.

As einman mentioned...your hydras ( and for me your Volvox sp!) alone make your micro aquarium sweet, Kurt!

Congrats on your indoor setup...love those hydras....sighhh, charlie guevara

[zzffnn]

^ If one wants a different size/dimension than that one, he/she may be able to DIY using polystyrene picture framing sheets available at home depot:

http://m.homedepot.com/p/Unbranded-11-i ... /100402512

Wall thickness of that sheet, per spec, is 0.05 inch or 1.27 mm. Optical quality should be decent to good, since it is made for picture framing.

[KurtM]

Kurt, do you think you can DIY something of similar dimension? I don't know if store-bought glass blanks can be very thin, if so, DIY should not be difficult.

What, exactly, are you asking about making again? In any case, hardware store glass runs about 2.25mm thick. Cutting to size and gluing it is no problem, very easy to do, I just watched a couple Youtube videos after searching "how to cut glass" and was on my way. Gluing is a simple matter of aquarium sealant from the hardware store, where you may also grab a glass cutter tool. So if 2.25mm thick window pane glass works for you, then everything you need is right down the street at the hardware store.

Since this is the first lab jack I've ever seen firsthand, I'm not sure what to expect to tell the truth. But there's no shaking, it's more or less rock solid as far as just sitting there goes. What I was disappointed in is smoothness of motion when turning the height adjustment knob (or more to the point, lack thereof). I need to take some time to disassemble it, and chase the threads and/or run some fine grade valve lapping compound through and put a polish on the threads, then it should be just fine.

Charlie G, I am also amazed that the hydras have lasted so long in the ~5 pint tanks. But volvox, not so much. Those rascals never live past four or five days at the most. If you (or anybody else) have suggestions to try for longer cycles, please post right up as I'm getting ready to re-establish a second micro-aquarium from a rich source of them.

[zzffnn]

Thank you, Kurt. You have answered all my questions very well.

2.25mm is too thick for me. I will probably DIY and use the cutter and sealant that you mentioned, together with the HomeDepot 1.27 mm polystyrene picture framing sheets. Or simply glass slide and transparency sheet for smaller vessels.

Your lab jack should work for my purpose. I won't expect a lab jack to move as smoothly as a microscope stage (in my rig, it does not have to move much at all). Pleas kindly let me know where you bought it and how much you paid. Thank you!

[KurtM]

I bought the lab jack too long ago for it to still be on the record, so can't tell exactly who I got it from except to to say eBay. About $40. Here's a close-up of what I got, which I repeat is nothing special. I think it'd help to apply dab of reel grease where the threaded rod enters the aluminum cross block, but not so sure I want a greasy threaded rod collecting dust and gunk. Top measures 5 1/4" x 6 1/2".

[gekko]

Secondly, I like the culture flasks, will order some soon. But I also need better open containers than the plastic and glass 50-60 mm Petri dishes I've gotten so far off eBay. Need better optical quality transparent surfaces for use on my inverted scope. Do you (or anyone else) know of better alternatives?

There are Petri dishes with flat glass bottoms designed for tissue culture for use on inverted scopes but they are expensive: see http://glass-bottom-dishes.com/pages/ The glass bottom is in the center of the Petri dish and is small (10 to 20 mm) and is labeled as having a thickness of No. 0 (presumably 0.08 to 0.13 mm thick), so very fragile, and thinner than the recommended 0.17 mm cover glass thickness for most objectives). Ted Pella sell another type with 0.17 mm glass bottom: https://www.tedpella.com/section_html/706dish.htm . But given your skill, you may want to try making your own by cutting out part of the bottom of a plastic Petri dish and gluing in a cover glass

[zzffnn]

I made a DIY version of the above, using 0.25 mm printer transparencies as walls with 0.6 mm thick internal space.

I tested its effect on image quality, using an aquatic insect and vorticella as test subjects. I think the "nano-aquarium" can produce high quality images with up to 10x NA 0.25 objective. Under 20x NA 0.4, image quality is not very good but still acceptable.

Thin wall and thin internal volume made lighting and photography easy. I could not get the same image quality with my smallest tissue culture flask.

So I am buying the above Carolina "micro aquarium". My DIY version is not durable enough to withstand routine handling, though its optical quality may be slightly better due to thinner walls and thinner internal volume. The Carolina one seems to have a good total volume, which I like for better sustainability.

If I only want best optical quality and have no cocern for sustainability, I would use sandwiched cover slips (vertically or horizontally).