When I checked on the cultures on Monday, the two that were dying looked like they had a bit of hope. I saw a bit of yellow slime mould in each, but one of them we overrun by another kind of mould. We did our best to scrape out the other mould and changed the oats in both cultures.
The slime mould finished solving the maze last night at around 7:30 pm. I got a picture of it at 4 pm, then forgot about it until 7:25 pm, when I checked on it and found that it had solved the maze, so I’m not sure exactly when it finished, but it was sometime just before 7:30 – maybe 6:30 or 7:00.
Today we transferred the filter paper from our second sclerotia attempt to an empty petri dish, cutting out sections that had other mould growing on them. We put that culture back into the incubator (along with the others) to let the filter paper dry out. We also changed the oats in the other living cultures.
Here is the original infographic I based mine on:
Here is my new and improved version: Big Mac VS Salad Infographic
And here is a link to the post with instructions for this assignment:
Today at 9:24 am we put the slime mould into the maze! We took a small piece of it with the agar it was on (from one of our first sub-cultures) and placed it upside down at the start of the maze. We put several oat flakes at the end of the maze. After watching it and taking photographs periodically every half hour, it didn’t look like it was growing much so we gave it an extra oat flake at 12:50 pm, thinking maybe it didn’t have enough food to be able to get all the way to the finish. It has grown minimally since this morning (less than a centimeter in the last 5 hours). Considering the top growing speed we read about online (1-1.8 cm/hr.), we are wondering if a) it didn’t have enough food or b) we didn’t put enough slime mould in to start with. My partner is planning to take it home and keep photographing it as time progresses. Hopefully it won’t grow too much during the night. The maze is quite large in comparison to the organism, so it could take a while.
We have been keeping the rest of the cultures in the incubator as they seem to grow better in there, and have started the process of making sclerotia again with our original culture. We changed the oat flakes in what was left of the original culture and put it back in the incubator to keep warm.
Leaving the cultures for the weekend seems to have been a good idea. The two that had formed crusty balls in the corners of the petri dishes don’t look any better (worse, if anything – at least one of them is growing another type of mould), but the others have grown and are looking better. The sclerotia we prepared on Friday (that has been in the incubator all weekend) to transfer back to its growing state has turned out well, except for a bit of contamination from some black mould, which we tried to remove this morning. We added oat flakes to most of the cultures and put them in the incubator to warm up and hopefully grow better since the room they are in is very cold today.
After discovering that a couple of our cultures didn’t look like they were doing to well this morning, we decided to put off the maze test until Tuesday. 2 of the cultures appear to have retracted their veins and formed small crusty balls near the edges of the petri dishes. Perhaps the clean-up job we did yesterday was a bit too much for them, but in any case, we want to allow the cultures the weekend to grow and recuperate before putting them into the maze we created.
Today we started the process of reviving some of our sclerotia. We selected a petri dish prepared with non-nutrient agar, sprinkled a few oat flakes on it, added a piece of dried filter paper containing sclerotia, and applied a drop of water to the filter paper. We sealed and labelled the petri dish, then covered it in foil to keep it dark.
A lot has happened since my last entry on January 23rd.
Here’s the rundown:
The incubator we ordered finally arrived on January 25th, and we tested it out for about 20 minutes.
The petri dish we had left out in the science room with only non-nutrient agar as a test culture dried up and resembled a warped sheet of very thin plastic.
On January 28th, we began our sclerotia-making endeavour. The first step to converting growing plasmodia into its dormant stage (sclerotia) was to put it on nutrient agar and incubate it. We prepared 6 petri dishes with semi-defined (nutrient) agar, transferring small bits of growing plasmodia onto 4 of them, leaving one out, exposed to the air in the kitchen this time as a test (like we did with the non-nutrient agar), and saving one for future use (and as a test) in our supply cabinet at room temperature. We exposed the test plate to the kitchen air this time instead of the science room air because we thought the air in the science room may have contributed to drying it out since that room is heated more. However, the results were the same: the agar dried up and formed a thin sheet of brittle dehydrated agar.
After preparing the petri dishes with the nutrient agar, waiting for them to set-up, lining them with filter paper, transferring the growing plasmodia onto the filter paper, sealing them with electrical tape, labelling them, and covering them with foil (to keep the cultures dark) then re-labelling the foil, we warmed the incubator up and put the petri dishes in.
We left the incubator on all night at its lowest setting (which turned out to be around 26-28 degrees Celsius), then turned it off the next day, waiting to see if the filter paper would dry at room temperature as we read it was supposed to.
Sometime after 24 hours, we opened our incubated cultures and transferred the growing slime mould that was still on its filter paper into new petri dishes that didn’t have any agar in them. We then put these back into the incubator.
On January 30th, we changed the oats in all of our growing cultures (the ones we were trying to make into sclerotia did not have oats) and looked at the original culture under a microscope. It looked….yellow.
We also came up with a couple of drafts for a possible maze, and are planning to test our maze-making skills tomorrow when my partner brings in some lego.
On January 30th, our plan for making the maze was to build it on a base piece and stick it directly into a Tupperware container we had previously purchased at the dollar store. But little did we know that on January 31st we would find out that the container featured an obvious yet hidden divider, making it unsuitable for out plan. Dunn dunn dunn dunnnn….
As it tuns out, the packaging of the container had conveniently covered the part where your could see the divider, so, upon purchasing the container, we didn’t know it had one. Therefore, the lego base piece didn’t fit into the container and we were left with another problem to solve.
We continued checking on the filter paper the next day or two to see if it was dry, but ended up finishing the sclerotia-making process shortly after we discovered another type of mould growing in one of the incubated cultures. Though the filter paper was not completely dry, the slime mould had formed small, dry, crusty clusters that were slightly darker in colour to the growing mould. We presumed these to be sclerotia.
We cut the filter paper that had the clusters and placed them in two other (plain) petri dishes – one for the transferred pieces that came from the contaminated culture and a separate one for the rest of them (prejudice, I know). We proceeded to cover these in foil to keep them dark, and they have been sitting undisturbed in the supply cabinet since.
When we transferred the migrating plasmodia that was on the filter paper from nutrient agar to empty petri dishes, we kept the supposedly ‘mould-free’ plates of nutrient agar that we used. Looking at them now, they are infested with other types of mould, and are quite interesting. Is that what we’re breathing in the air all the time? I guess those plates display the difference between non-nutrient and nutrient agar: one is easy to keep unwanted things from growing on (the non-nutrient agar) and the other is not. Or so it would seem.
It is interesting to note that the petri dish of nutrient agar that we sealed immediately after preparing and have not re-opened since has not grown anything, So how did the other mould get onto the now slime mould-free plates? Some things might have to remain a mystery, but they didn’t just appear out of thin air …actually, on second thought, they might have.
On February 4th we attempted to sterilize a few things….first some used petri dishes, then the lego we would use for our maze, We soon discovered that a) petri dishes cannot withstand boiling water and b) lego doesn’t do much better. After first trying the petri dishes and ending up with warped tree ornaments, we thought we would be smarter with the lego and try a test piece. Unfortunately we discovered a little too late that the two-blocks could withstand boiling but the others could not. Our lego blocks and base piece now not able to be attached together again, not to mention being slightly discoloured, we had to think up another plan of action.
Our new bright idea (let’s hope this one turns out better than the last one) was to get a Tupperware container’s undivided attention (or, rather, just a Tupperware container without any division at all), that would fit the size of the maze, put a layer of non-nutrient agar on the bottom, and stick the lego pieces directly into the agar when it was semi-solidified. We then embarked on the now somewhat habitual task of melting and pouring the agar.
My partner took care of fixing the lego pieces into place with only minor complications. The only things that went wrong were a) the agar solidified a little too much before the lego places were placed in it, resulting in small cracks, and b) there was a slight change in formation as far as the shape of the maze (apparently my drawing skills could use some improvement).
With a few re-calculations today (due to the slight change in shape of the maze) on how long the mould would take to solve the maze if it grew at top speed, we came up with an estimated time of maze completion being about 10.2 hours. However, we are expecting the mould to take longer than that, as that is the top speed we read that slime mould grows (1.8 cm / hour), and another source said only 1 cm / hour.
One of us is planning to take the mould home while it is solving the maze so we know exactly how long it took to solve the maze.
Looking at our original culture we discovered a bit of contamination from some type of black mould. We removed the chunk that was contaminated and soon discovered that our other cultures were growing out of their petri dishes. Looks like the taping job we did didn’t quite suffice, and the mould must have been hungry because it was making it’s way down the sides of the petri dishes, probably looking for food. We washed the lids of the petri dishes, cleaned up the sides and bottoms of the containers, and re-taped them – better than last time. We also changed the oat flakes that the cultures were eating, giving them fresh ones and trying to put them in the centre this time to encourage the slime mould to grow into the middle of the petri dishes rather than toward the outside.
I think that about sums it up! Tomorrow we put some mould in the maze and see how it goes!
1. Open your Portfolio Audit Inventory sheet word file
(Make sure it’s already in your Dropbox folder)
2. Place the evidence document you want to link into your shared Dropbox folder
(Do this by going to Dropbox in your folders section rather than online at www.dropbox.com)
- You may chose to organize your documents into folders at this point (if you move them later the link will not work properly)
3. Go to Insert in your inventory document, then click the Hyperlink button
4. Choose the file in your Dropbox shared folder
- Click the Browse for File icon near the right side of the window (it has a picture of a folder opening on it)
Select the file you want and press OK
5. Test the link (Ctrl+Click) to make sure it works
1. Go to www.dropbox.com
2. Sign in
3. Click on Sharing button on left-hand side
4. Click on New shared folder (near right-hand side)
5. I’d like to create and share a new folder should be pre-selected
6. Name your folder with your first name and last initial.
7. Click next
8. Invite collaborators to this folder
Type in the email address(s) of the person/people you want to share the folder with.
9. Click Share folder
You should now see the new folder in your Dropbox under Sharing
10. Move your Portfolio Audit Inventory sheets into this folder as well as the documents you will use as evidence for your portfolio
If you already have your files in another folder, simply cut and paste them into your new shared folder, then you can delete the old one.
a) your evidence documents are in the folder
b) they are organized
before you link them in your inventory sheet. If you do either of these after, the links will not work properly.
On Monday we successfully sub-cultured 5 plates of physarum polycephalum.
We started by melting a bottle of 2% non-nutrient agar in a pot of water. We had to start of with the water and the bottle of solid agar at the same temperature, gradually warming them up so the glass agar bottle wouldn’t break. We also unscrewed the bottle cap most of the way so air could get in and out. Once it was fully liquefied, we took the bottle out and waited for it to cool enough to pour. We then poured the agar into 5 petri dishes, opening them only long enough to pour the agar in to avoid contamination. We kept the lids on for about 30 minutes while waiting for the agar to set, then prepared to sub-culture from our plate of growing mould.
We ended up having to sterilize the X-Acto knife blade three times (it got dropped, etc.) due to worrying we might contaminate the mould – kind of ironic if you ask me. Once the agar was set and we were ready to subculture, I carefully cut 5 small cubes from our growing culture, placing them upside-down into the petri dishes. We took 2 squares with the oat flakes they were already eating, 2 squares without that we placed directly on top of oat flakes, and 1 that we placed with no oat flakes directly touching it.
From the results so far, the ones we took with oats they were feeding on are growing the best, followed by the ones we put directly on top of an oat flake. The one we took by itself that was not originally touching an oat flake has not grown as much but is still alive and well, from the looks of it.
My partner also replaced some of the oat flakes our original culture was eating because they looked like they were either a bit mouldy (with a different type of mould) or might soon get mouldy, as we read that physarum polycephalum will leave old oat flakes for new ones, which can lead to older ones getting contaminated by other fungi.
Yesterday we prepared another 10 petri dishes with plain 2% agar for future use, putting 5 of them at room temperature and 4 in the fridge. The last one we left out, with the lid off in the science room so we could see if anything (that might land on it from the air) grows on it. So far nothing has, and considering how well the mould is doing it seems less finicky than we originally thought. The non-nutrient agar is working great, and the mould is growing just fine!
Today we put one of the 5 sub-cultured cultures into the fridge so we can see how much impact the cold has on its growth speed. By comparing it to the others, we hope to determine if we can leave it in the fridge over the weekend without it growing too much.