Deep Space said:

Anomalocaris said:

Lets see who wins the pics contest

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I’ll play 🍻

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Anomalocaris said:

Its ok when its in a three way

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Twigg said:

Hello ladies and fellas,

I just put some rather thin and whispy pan. J
thats been in my fridge for around 6 years to some fresh agar.

If anything happens ill make a separate post for it. Wish me luck, ima need it 😆

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fahtster said:

There’s not much of an aerial leading edge on that.  A cabin is probably a good idea but once you start taking aerial pulls, I wouldn’t go back to a cabin.  The only thing I’d incorporate would be a trench.  Simply because it’s taking away plate media.  I’d stick with aerial transfers until the culture is clean or at least clean enough (for you) to put out a decent flush. 

These are bvi.. this is the state of a culture where I say “now we’re gettin somewhere”



I can tell it’s still dirty by how there’s an area around the transfer that’s more barren than the better looking outer/leading edge

Here’s another plate that is slightly cleaner than that one..



Notice how the barren area around the transfer is much smaller.  The idea is to make that area as small as possible (or nonexistent) until the culture looks like there’s little to no difference from the leading edge to the center of the transfer and all the myc is aerial like this




If you look at the first pic in each of those groupings above, you can see what the dirtiest, cleaner, cleanest looks like..

… …

Now, will those first plates flush out?  Probably.  Will the first couple flushes suffer a bit?  Probably. 

If a lot of ppl are gonna start doing AM transfers, I wanna say some things that have really helped move things along for me.  First, if you are using a hooked scalpel, make sure you don’t push the tip down too hard into a hard surface after flaming red hot.  It’s a very subtle amount of pressure.  If you push too hard, you’ll end up curving the tip back towards the handle and it’ll be very hard to get the tuft off the hook. 

I made a drawing to demonstrate the do and don’t.


Something that I think is really important is that you don’t want to keep jabbing the scalpel tip into the receiving media plate trying to get the tuft off the tip of the blade.  If there is bacteria on the tuft, all you’re doing is spreading that bacteria all over the area surrounding where the tuft eventually ends up.  This is doing you no favors.

What I started doing a while ago that’s really been a game changer for me in that regard is to use an extra firm (3.5%) water agar plate to initially dislodge the myc tuft onto the firm agar surface and then once it’s off on the WA, all you have to do is basically touch the scalpel tip to the tuft and it’ll barely stick to it.  Once it’s on the tip again, you can take it to the receiving plate and it very easily comes off the second time.  I can basically just touch the dangling tuft to the surface of the receiving plate and it sticks right there without the scalpel even touching the surface.



It becomes just like placing a wedge.  This really gives the myc (that outnumbers the small amount of bacteria that may be on it) a chance to break away in all directions and outrun the infection much quicker than if you planted bacteria all around it by jabbing the tip into the surface which really negates the whole point of the aerial transfer. 

Here’s the firm water agar plate after a transfer session.  For each transfer I find a new unused portion of the plate to dislodge each tuft



That’s a great vid by kloakk, btw.  The one note I have is that you don’t want to take the transfer from the barren section of the growth.  The furthest leading edge possible is where you want to grab from.  Grabbing myc from the inner portion more than likely isn’t going to move the culture forward towards clean (unless it already is clean, ofc)

Here’s a plate I did a couple AM transfers from.. there’s one by the number four that’s less obvious



A side note about that satellite bacteria colony… that type of bacteria stands no chance against pan myc.  The bacteria is fairly slow compared to pan mycelium.  It’s non-motile. It’s just chillin, living its best life and I don’t even worry about those kinds of bacteria fackin up my cultures.  The only time I’d be worried is if I had water rollin around my plates.

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fahtster said:

If a lot of ppl are gonna start doing AM transfers, I wanna say some things that have really helped move things along for me.

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Great stuff as always Faht, appreciate the effort of the work as well as sharing it with the rest of us

fahtster said:

Something that I think is really important is that you don’t want to keep jabbing the scalpel tip into the receiving media plate trying to get the tuft off the tip of the blade.  If there is bacteria on the tuft, all you’re doing is spreading that bacteria all over the area surrounding where the tuft eventually ends up.  This is doing you no favors.

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I am not in disagreement, but I would like to know what you do IF the transfer doesn't come off without dipping the scalpel in the agar

Do you:
(a) Call it a loss, sterilize the scalpel and get a new AM transfer.
(b) Dip the scalpel once and if it doesn't stick call it a loss, sterilize and get a new transfer and plate.
(c) Get the transfer off, but do more AM to new plates until you get one that went perfectly.

I ask because if we only go for perfection, I at least, will burn through a lot of plates in a short time, would you say that any AM transfer that isn't perfect is no better then a regular wedge transfer?

Quote:

fahtster said:That’s a great vid by kloakk, btw.  The one note I have is that you don’t want to take the transfer from the barren section of the growth.  The furthest leading edge possible is where you want to grab from.  Grabbing myc from the inner portion more than likely isn’t going to move the culture forward towards clean (unless it already is clean, ofc)

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Again, not in disagreement here either, and I try to get the AM from the leading edge, the video was made in open air with a fully colonized plate pulled out the trash pile

My question here is, if there is bacteria on a plate, would getting AM transfer be as dirty as getting a wedge transfer? I ask because sometimes the leading edge is flat on the agar while the barren section is supporting each other and doesn't flatten out in front of the FFU.

In other words, would AM transfer from the barren section be as dirty as a wedge transfer from the leading edge?

Quote:

I am not in disagreement, but I would like to know what you do IF the transfer doesn't come off without dipping the scalpel in the agar

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Well, I think if the scalpel dips in the agar and the transfer is left there, there’s not really any difference because the scalpel went through the same parts as the tuft.. they’re both as dirty as each other.

From your video, it didn’t look as tho you had a hard time getting the tuft off.  My hands aren’t steady at all so using a scalpel without a hook doesn’t work very well for me.  I rest the scalpel on the edge of the plate sides so I don’t dip into the surface when I’m taking the transfer which works great but the tuft likes to get a bit stuck onto the hook.  This creates a problem putting it on the receiving plate where I have to use the plate surface and end up jabbing multiple spots on the surface to get it off and sometimes it just disappears and I don’t even know if I got it off or not

For example.. here’s a plate where that happened and more times than not, the transfer isn’t successful


Chances are that it was a pretty tiny tuft to begin with and after trying to remove it on the surface, it compacted into something I couldn’t even see on the tip anymore.  And all this is because I can’t seem to get my gellan gum as firm as my agar.  That’s why I use a firm WA plate to dislodge the tufts initially.  I can use the WA to jab as much as I need to get it off but it usually doesn’t take much with a nice firm surface.

I noticed a while back when I was using agar that every once in a while, the tuft would come right off and those transfers were always the best transfers so, what I started doing was using part of the receiving plate off to the side as the area is dislodge the transfer and then move it over to the other side of the plate where there weren’t a bunch of stabs in the surface.  Then I just decided to make a bunch of firm WA plates that I use each transfer session so my receiving plates didn’t have a bunch of bacteria all around it. 

All that’s to say, I don’t want to waste a bunch of plates either   the only time I call a plate a loss is if I drop it.  Aerial transfers are always going to be better than wedge transfers (to me) even if you do end up jabbing the plate a bunch or don’t do a “perfect” AM transfer.  But really, the throw away firm WA plate makes every transfer “perfect” because all the bullshit that might happen is left on the WA plate. 

Quote:

“ Again, not in disagreement here either, and I try to get the AM from the leading edge, the video was made in open air with a fully colonized plate pulled out the trash pile

My question here is, if there is bacteria on a plate, would getting AM transfer be as dirty as getting a wedge transfer? I ask because sometimes the leading edge is flat on the agar while the barren section is supporting each other and doesn't flatten out in front of the FFU.

In other words, would AM transfer from the barren section be as dirty as a wedge transfer from the leading edge?”

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Kinda figured that and you were just showing what an aerial transfer was in a general sense.  The “note” wasn’t really for you, more the ppl watching the video that haven’t done it before… a further clarification.

I’m so anti-wedge transfer lol. At least for pans.  Any AMT is better than a wedge, imo/e.  Even if it’s from the more inner part of the culture… it’s more than likely not going to get it cleaner, but it’s probably not going to make it worse either. 

What I will do in that situation (and ik exactly what you’re talking about because it happens to me too) is stop if the leading edge get blown down because if the leading edge is healthy enough, you can wait a couple few days and that edge will pop back up and you can take another stab at it. 

This actually happened to me the other day.. I thought this culture would be a super easy AM grab and when I opened the plate, the myc was so delicate, it immediately started laying down and I wasn’t fast enough to get a tuft to grab into the hook.  I kept dipping into the surface and eventually said fuck it and took a leading edge wedge because I thought it looked clean enough that would be safe to do


Time will tell but anyway, you can see how the leading edge grew back in those few days even better than it was.. possibly from the influx of o2 from the plate being opened

I’m glad you asked that question because I forgot about that plate and will do an AMT from the leading edge tomorrow now that it’s grown back

I’m not saying I have all the answers but I’m doing a fuck ton of AM and just sharing what I’m finding along the way