I didn’t get a chance to go in to lab this weekend so I couldn’t get my notes, so I’m doing it now. I posted all my results from looking under the scope on Friday evening here. Side note: I looked at the sample again today and everything was still busting with life. No tardigrades, but I got some cool videos which I’m posting to BenchFly (I’ll post in my notebook when they’re up).
Lichen in petri dish with DI water added to add moisture
When we first gathered the samples, I added water to the dry samples. The lichen sample got about 10mL of DI water and was allowed to soak for a few days.
I filled 3 analyslides (see Experiments’ product list link above), each with 6mL of DI water.
I would use a pair of tweezers to grab a chunk of lichen from the original sample and place it in an analyslide. With the tweezers I would shake the lichen sample for about 1 minute. This would break the lichen into much smaller chunks which should release the water bears from their shelter/slumber.
lichen in analyslide suspended in di water.
In the lab we have an Olympus IX71 inverted microscope and I used a 20x objective to visualize the samples.
I ended up checking 2 of the 3 samples I prepared, and saw nothing but lichen chunks in there. Eventually I looked at the original petri dish sample and got all the cool results that I mentioned earlier.
Mr. (Dr?) Shaw left me a comment the other day directing me to some material he wrote regarding finding tardigrades. On his site he wrote an article that is similar to the one I linked in the this post. So on his post I left a comment and he responded. You can find the transcript in that link above, but if you are too lazy to click then I’ll just write it out here:
From me: How many can you typically find in a 10mm by 10mm clump of lichen or moss? Are they plentiful or kinda solitary? Thanks for commenting in my open scientific notebook!
From Mike: You are lucky if you find two or three in any sample. Typically, I would scrape lichen into a small paper coin envelope. I’d make a suspension in bottled or filtered (Aquafina or Poland Spring) water in a plastic petri dish and let it sit overnight. The next day, I could spend maybe 15 minutes going though the sample under the microscope, and if I found even one tardigrade I would consider myself lucky. Often, however I would find two or three, or some eggs. I have had very little luck with moss. It’s got a lot of sand particles to sift through, and moss itself is hard to look through. Like looking through a jungle for a hamster.
The expression is: Your first tardigrade is the hardest to find. Good hunting! Mike
Following some “protocols” I’ve read about finding tardigrades, I began by taking some lichen samples and shaking them vigorously in water. The hope being that you spring the little guys from their shelter into the free water. Honestly I had no expectations and no idea what to expect. While I don’t think I was successful today, I did have a ton of fun seeing the amazing amount of life in just that little bit of lichen.
I’ll post my notes about the extraction tomorrow. I took written notes because I was in a time crunch and I didn’t have time to transfer them here, and my lab notebook is in lab and I am home. So tomorrow then. But here is my quick protocol from what I remember:
Use tweezers to grab a piece of lichen.
Put 6mL DI water in analyslide and shake lichen in water.
Seal slide and do two more times.
I looked at one of the samples under a 10x objective and almost did a second one, but I got too excited and wanted to see what was going on in the original petri dish so I ditched the second sample and looked at the original. The first sample revealed nothing but a lot of lichen chunks. The petri dish however was full of life. Tons of little microbes running around in the sample, and they move FAST!!! Here are some awesome pics of whatever I could capture.
Amoeba thing again embedded in the blob. The lichen chunk seemed to contain something else that was alive cause it was pulsing, but maybe that was the amoeba poking it.
Tiny amoeba like thing. Moved quickly but liked to be near the lichen blobs.
Another view of the squirmy tardigrade like thing. For a while it was tethered to that blob, but broke free and didn’t go anywhere.
Is this a water bear? I don’t think so, but it was squirming around and I got really excited. It is as big as the paramecium thing.
This thing is huge! It was at least 10x as long as most of the other amoeba/paramecia type things in the sample.
I’ll look at the samples more tomorrow, since I’m not worried about the tardigrades dying, because they are the FUCKING TERMINATOR SENT FROM THE FUTURE TO DESTROY US ALL!!!! Seriously though, those things are impossible to kill, which is why I think putting them in D2O (heavy water) is a worthwhile venture.
Here are some observations from the banks of my memory:
There are a lot of microbes in this little tiny amount of lichen. That makes me fear for my life to just go outside. What the hell is growing in my home compost? I think I want to live in a bubble for the rest of my life now. It makes me even more scared that I bite my finger nails, those things are in my belly!
The size range of things that are alive in this sample is amazing. There were things that were super tiny (even smaller than the tiny amoeba thing) all the way up to that giant paramecium type thing.
Yes amoeba and paramecium are the only two microorganisms that I know about. And now tardigrades. Thank you high school biology.
The small things would move around really fast. And the large things would move a bit slower, but they could still move faster than I could. And at this size scale, brownian motion is of almost no affect. I didn’t witness anything diffuse via jiggling. It was all controlled motion, and things that were stationary were completely stationary with the exception of whenever the petri dish moved, that made everything in the sample swish around ever so slightly, sorry for the ridiculously long run on sentence I promise I won’t let it continue for much longer, and now I’m done.
This sentence will be short.
Lichen all by itself is pretty cool to look at. Next time I’m in the lab I’ll take way more pictures and show you.
I feel like DIC microscopy would be very helpful here, but I didn’t have time to set it up.
I will also take some movies of stuff to put here. Benchfly? Sounds good to me!!!
Even if I find no tardigrades, I’m super excited to get back out there and find more samples!!
I know there is a lot that I’m not remembering. I felt like I was in the movie “The Abyss”:
I’ll take much more detailed notes next time when I can dedicate hours to just staring into the world of the itty bitty. Plus I’ll get a couple of objectives of different magnifications so that I can get in closer to this stuff. I briefly tried our 60x objective, but the petri dish was too thick. Anyone have a 20x objective or a 30x or something more than a 10x but less than a 60x? Thank you.
In summary, hooray for me! This was super exciting and I can’t wait to explore some more. By the way this is the cool stuff about science that doesn’t get to go in a journal publication. But here in my notebook I get to say how awesome it is, how much fun I have, what I did, and I get to show emotion! Win for open notebooks if you ask me!!!
I received a comment from a one Mike Shaw who directed me to a document about tardigrade finding and observing. You can find the document here (be warned, the link is a .pdf and it may be a bit large). His article is first and has some interesting information.
Alex wrote up a nice post that I will not replicate here. She discussed our adventure and our plans for the cute little bundles and she even found a site that discusses how to extract the little guys for observation. Finally, she posted a picture of the samples that we took.
Great post and tomorrow we’ll work on extracting some tardigrades. I’m kinda optimistic about the lichen, but not so much the other stuff. We shall see…
As I said, yesterday Alex and I went for a walk to look for tardigrade habitats. If you are unfamiliar, tardigrades are microscopic animals that are super adaptable and can survive in nearly any environment on earth and are even capable of surviving in space. They are more commonly called water bears but they won’t swipe your picnic basket (or at least not that I know of). In my (very limited) research, they are found to rent duplexes in lichen and moss, but I’m sure they can be found in other exotic locals. Because of this I decided to focus my search on that.
I’m no ecologist so my knowledge of where to find lichen and moss is pretty limited. I know moss needs water and lichen is more rugged, but both like moistness and dampness (maybe those are the same things) so I figured near the river I would find an abundance of this. NOPE!
After walking for a little while, I decided to get a sample of the river and maybe we could find some other microscopic organisms. I also got a sample of some mud, because why not. We found a dead tree too that looked like it had moss on it, but it turned out to be some weird fungal growth, so we took some of that as well.
On the way back to Alex’s ride we found a tree that had a ton of lichen on it. I tried climbing it but it had all these yucky bugs on it. Alex insisted they were termites, but I assure you (and her) that they are not. Just because a billion of them lived in a tree does not mean they are termites. Anyways not only were the bugs a nuisance but I couldn’t get a decent grip. Luckily Alex found some nearby trees that had lichen growing on more accessible regions. So we took some samples of this as well.
I’m optimistic now.
Below you’ll find pictures of the places we took our samples from, and below that you’ll find a google map I made of our adventure.
We’ve had some gorgeous weather lately here in sunny NM and today it was supposed to get up to 72F (it never did though…) so I planned on going hunting for Tardigrades. I don’t know if I was successful, but I did get some promising samples. I’ll go into more detail later because I’m waiting for some pictures from Alex’s phone to be emailed, but it was a pretty fun adventure.