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Josex
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: BrownBear] 2
#27053746 - 11/23/20 10:52 AM (3 years, 2 months ago) |
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60ml syringes wrapped in foil and a pp5 test tube full of 14G needles wrapped in foil, all sterilized inside a gallon jar.
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D3_Myc
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Josex]
#27053765 - 11/23/20 11:04 AM (3 years, 2 months ago) |
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This is all good to know, currently I’m working with just pouring in front of the flowhood but planned on making a few lids with injection ports and getting some syringes. Is there a favorite brand / size of syringe? (I’ll utilize the search button as well)
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mushhead
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: mushhead]
#27054499 - 11/23/20 07:23 PM (3 years, 2 months ago) |
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Quote:
mushhead said:
   Left is B+ Right is APE. LME heat dissolved into distilled water with a 45min PC cycle. Some sediment but no other microbes save mycelia in here.
 B+ (Left) APE (Right)
  A couple other LC testing nute levels.
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Mateja


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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: mushhead]
#27054525 - 11/23/20 07:41 PM (3 years, 2 months ago) |
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From my limited knowledge about those broths (only knowing these pics and what you've described so far) I'd say B+ looks turbid APE looks clear. But it's worth remembering that discernment comes from having the collective data on everything from time of inoculation, temp, changes in appearance no matter how slight, timing of the changes, many many small factors that I myself know very little about I'm totally a baby just discovering as I go.
I'm just pointing out that the one factor we have that appears to be a very rigid measuring device (the turbidity phenomena itself) nevertheless eyes can also be tricked, I've already been punked by some broths a few times, so observing them regularly in the hours/days after inoculation tends to provide the most valuable data ime.
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Pastywhyte
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27054535 - 11/23/20 07:47 PM (3 years, 2 months ago) |
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I like these, they look ready to rip into some grain.

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mushhead
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Pastywhyte]
#27054542 - 11/23/20 07:52 PM (3 years, 2 months ago) |
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Quote:
mushhead said: I've been working on my LC nute levels. Trying to get them as clear as everyone's here. My LCs are clean, but have sediment because I've been experimenting with nute levels. My first batch is 5g LME to 500ml distilled water. LOTS of sediment. Second is 2.5g LME to 500ml distilled water. Medium-minor sediment. Third is 1g LME to 500ml distilled water. Mostly none. Small amount visible on bottom of jar. Fourth is .5g LME to 500ml distilled water. Pretty clear. Pics after I finish a bunch of errands today.
I PC for 45min each batch of two jars I did. The fourth batch is just the one jar. They're in succession from first batch to fourth which I'm sure you can tell which has the highest nutes. You see I believe that the B+ jar isn't contaminated at all and the turbidity comes from the amount of nutes used which is like 5~6g because it was my first one after trying with Gulf coast which worked amazingly well for me. EDIT: These jars were kept at 22c consistently without deviation.
Edited by mushhead (11/23/20 08:15 PM)
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mushhead
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Pastywhyte]
#27054558 - 11/23/20 08:02 PM (3 years, 2 months ago) |
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Quote:
Pastywhyte said: I like these, they look ready to rip into some grain.


Pasty I aspire to be upon your level immensely.
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Lemgrub


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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: mushhead]
#27054565 - 11/23/20 08:06 PM (3 years, 2 months ago) |
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Got a APE syringe in today, first time working with it. But it looks like they germinated...
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Josex
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Lemgrub]
#27054581 - 11/23/20 08:16 PM (3 years, 2 months ago) |
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The pic isn't good but Ape spores kinda look like that. If they take time to show growth on agar (and they usually will) you can be sure it's spores.
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Mateja


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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Lemgrub]
#27054582 - 11/23/20 08:17 PM (3 years, 2 months ago) |
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Quote:
Lemgrub said: it looks like they germinated...

I'm afraid "they" could mean 'several types of organisms' germinating. Both fungus myc and bacteria is what the pic says to me. I wonder if it'd be possible to salvage a part of the myc in there tho, maybe to dilute a small part heavily in sterile water and then to extract a samle to put to agar? What do y'all think about this guys? I'm sure someone has tried something similar before, to cleaning out myc from a bacterial broth lol
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Lemgrub


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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27054625 - 11/23/20 08:44 PM (3 years, 2 months ago) |
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About as good a pic as I can get. looking closer I can tell there are spores, but it looks a bit more viscous than usual. I nocced some agar and brf cakes and a glob came out in one of the cakes. I've looked at plenty of spores under the scope and I've seen a large mass of spores sort of suspended in a matrix of hyphal cells, I don't know if they're just dead in some way or if they were sterile monokaryotic mutant lineages or something like that.

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Pastywhyte
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Lemgrub]
#27054658 - 11/23/20 09:28 PM (3 years, 2 months ago) |
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Looks like APE spores to me.
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Lemgrub



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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Pastywhyte]
#27054730 - 11/23/20 10:49 PM (3 years, 2 months ago) |
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Tight
Here's a tissue needle biopsy LC I started 11/13 from Colombian Rust. The liquid has retained the same clarity throughout. Believe it's growing somewhat slowly due to higher sugar content, 4%
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Josex
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Lemgrub]
#27054744 - 11/23/20 10:58 PM (3 years, 2 months ago) |
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Are you swirling it? That will break up that glob and speed up colonization. It's being terribly slow regardless, I would not trust it but you can always test it.
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Mateja


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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Lemgrub]
#27054790 - 11/23/20 11:49 PM (3 years, 2 months ago) |
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Quote:
Lemgrub said: Tight
Here's a tissue needle biopsy LC I started 11/13 from Colombian Rust. The liquid has retained the same clarity throughout. Believe it's growing somewhat slowly due to higher sugar content, 4%

I've done side by sides with clones before and a varience in nutrient density didn't seem to affect growtht speed much if at all. At least not with the cultures I tested. The myc colony in your LC is far to small for a 10day old LC, it should really be close to being fully colonized at this point. This raises some concerns. Also (and I could be wrong) but from the pic it doesn't look like the light source is very close to the jar so it's hard to determine how clear/turbid that broth is. The brighter the light is shining on a turbid broth the more turbid it will appear and the less light that is being diffused inside the broth the less turbid it will appear so the amount of light shining through will hugely affect how the broth is perceived ime. And lastly I have to say that personally I can not come up with a reason to use stronger broths than 0.1%.
Adding a massive amount of particles to a LC like 2-4% malt will make it that much harder to inspect since you don't have a 'default' like a crystal clear broth for example in which case you're able to notice the slightest increase in particles (since the broth goes from 'clear' to 'not clear') but with a broth with already tons of particles in it then it's simply not reasonable to assume you'd even be able to notice a change in amount of particles (relying on noticing the change from 'turbidity' to 'more turbidity'. Hope that helps. Gl
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Edited by Mateja (11/23/20 11:53 PM)
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sonoramo
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Pastywhyte]
#27055222 - 11/24/20 09:53 AM (3 years, 2 months ago) |
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Quote:
Pastywhyte said: I like these, they look ready to rip into some grain.


Is the brownish stuff on the bottom sediment? If it is, it seems like a lot of sediment. Like, if you swirled the jar it would make everything look cloudy. Was it there from when you prepared it, or did it precipitate out while the culture grew?
Finally, what recipe did you use for the broth?
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Lemgrub



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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Mateja]
#27055238 - 11/24/20 10:03 AM (3 years, 2 months ago) |
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I gotcha. The (very bright)lamp was probably an inch from the cup. I've had a few LCs contaminate since making this and the difference is stark. I don't believe this one is contaminated to this point.
And as far as the solution I used, I quickly looked at a tek that recommended 4%, so I went ahead and did it since the PC was almost ready to go. Seemed high and further reading supports that. And honey isn't too bad particle wise.
Josex, yes I've been swirling a couple times a day. Finally a few days ago some myc broke off from the biopsy and there's separate colonies now. I think the colonization will speed up now.
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Pastywhyte
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: sonoramo]
#27055256 - 11/24/20 10:08 AM (3 years, 2 months ago) |
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Quote:
sonoramo said:
Quote:
Pastywhyte said: I like these, they look ready to rip into some grain.


Is the brownish stuff on the bottom sediment? If it is, it seems like a lot of sediment. Like, if you swirled the jar it would make everything look cloudy. Was it there from when you prepared it, or did it precipitate out while the culture grew?
Finally, what recipe did you use for the broth?
The brownish stuff on the the bottom is a giant puck of solid agar. The “broth” is simply sterile water added to the vessel once the colony on the agar has grown out and looks healthy. Use whatever recipe you like, this was just undiluted grain soak water and 2% agar for the puck.
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coversall
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: Pastywhyte]
#27055286 - 11/24/20 10:23 AM (3 years, 2 months ago) |
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Chucked an agar wedgelet into some LME broth today... It floated... I-is that ok?
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D3_Myc
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Re: LC Training Camp: Interpreting Visual Cues To Predict The Quality Of A Broth [Re: coversall]
#27055318 - 11/24/20 10:47 AM (3 years, 2 months ago) |
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Started some enigma on LC last night. T1 from a hot pour (it wasn’t needed just wanted to try it) This is one in a series of side by sides I’m going to try with grainwater and lme. My hypothesis is that the enigma will be more aggressive in grainwater based on its growth patterns on agar vs when it goes to grain. I’m looking forward to seeing the results
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