|
 fyouseeku
Chemist
Registered: 05/25/06
Posts: 79
|
 Best way to make an LC 
#6594733 - 02/21/07 04:42 PM (5 years, 3 months ago) |
|
|
What is the best way to make an LC of gormet mushrooms?
I am use to using kero for other types of mushrooms but still can not find out about the gormet kind.
Thanks 
|
 RogerRabbit
Bans for Pleasure
Registered: 03/26/03
Posts: 36,974
Loc: USA Mountain Northwest
Last seen: 12 hours, 29 minutes
|
Re: Best way to make an LC [Re: fyouseeku]
#6595007 - 02/21/07 05:50 PM (5 years, 3 months ago) |
|
|
Obtain a pure culture on agar. Test with a microscope to make sure no bacterial rods are present in the dish. Drop the contents of one dish, agar and all into 1 cup of water in an eberbach and blend with three short blasts of no more than two seconds each. Pour the slurry into a flask with 1 liter filtered water, 20 grams malt extract, and 2 grams yeast. Stir 24/7 and use within 7 to 10 days to inoculate grains.
RR
--------------------
www.mushroomvideos.com
semper in excretia sumus solim profundum variat
|
Cryogenicz
what?
Registered: 07/01/04
Posts: 2,344
Loc: Oregon
Last seen: 1 day, 13 hours
|
Re: Best way to make an LC [Re: RogerRabbit]
#6595715 - 02/21/07 08:24 PM (5 years, 3 months ago) |
|
|
which type of microscope would you use to check the petri dish?
Link me!!!
--------------------
www.MycoPath.com
Mushroom Spawn, Cultures, Fungi Bags, Casings, Master Grain Jars, Bags for In-vitro, Laboratory supplies, and much more!
Mushroom Supplies. Fast Turnaround Times. Great Service.
orders@mycopath.com
enter code shroomery for 10% off product.
www.FungiForum.com
|
RogerRabbit
Bans for Pleasure
Registered: 03/26/03
Posts: 36,974
Loc: USA Mountain Northwest
Last seen: 12 hours, 29 minutes
|
Re: Best way to make an LC [Re: Cryogenicz]
#6595829 - 02/21/07 08:41 PM (5 years, 3 months ago) |
|
|
You'd want a light microscope. You can see bacterial rods very well at 400X. They look like little pencils. A halfway decent starter microscope would be this one http://www.microscopesusa.com/Observer_III.html
If you can afford a better one, check out the lab microscopes. Look for the largest diameter objectives you can afford, because the larger the objective, the more light it takes in. You don't need 1000X so don't worry about getting a starter microscope with the 100x objective.
RR
--------------------
www.mushroomvideos.com
semper in excretia sumus solim profundum variat
|
FreeSporePrints
Mycelium Jedi
Registered: 03/06/05
Posts: 1,130
Loc: Rome, ITALY
Last seen: 6 days, 9 hours
|
Re: Best way to make an LC [Re: RogerRabbit]
#6596305 - 02/22/07 12:06 AM (5 years, 3 months ago) |
|
|
I quote RR and remember to discard the edges near the uncolonized agar media AND the uncolonized agar media.
I've a NOVEX stereomicroscope, model P-20, i've never used it because i reach just the 80x and for now i haven't money to buy new powerfull lens.
These stereomicroscope are really usefull because you can put directly the petri on the "platform", beyond a flowhood. In this way you can provide a good sterility for your culture.
Fabio
--------------------
Edited by FreeSporePrints (02/22/07 12:17 AM)
|
RogerRabbit
Bans for Pleasure
Registered: 03/26/03
Posts: 36,974
Loc: USA Mountain Northwest
Last seen: 12 hours, 29 minutes
|
|
80X is about as powerful as you're going to get in that type without moving up to a seriously expensive scope. I use mine mostly in the 15X to 25X range because image is so much clearer. You'll need a light microscope to see bacterial rods. With a light microscope, the light is below the object to be viewed, and you see the light that shines through it. With a stereo or zoom microscope, the light is on the same side as the lense, so you see the light reflected back from the object. Here's the one I use. I like the boom stand because I can swing it over in front of the flow hood. I also have two more boom arms to extend it out farther, but usually just keep the shortest one attached because there's less movement and bouncing.
RR
--------------------
www.mushroomvideos.com
semper in excretia sumus solim profundum variat
|
Cryogenicz
what?
Registered: 07/01/04
Posts: 2,344
Loc: Oregon
Last seen: 1 day, 13 hours
|
Re: Best way to make an LC [Re: RogerRabbit]
#6598375 - 02/22/07 04:09 PM (5 years, 3 months ago) |
|
|
What would you consider an intermediate microscope, which could view bacteria?
-Graham
--------------------
www.MycoPath.com
Mushroom Spawn, Cultures, Fungi Bags, Casings, Master Grain Jars, Bags for In-vitro, Laboratory supplies, and much more!
Mushroom Supplies. Fast Turnaround Times. Great Service.
orders@mycopath.com
enter code shroomery for 10% off product.
www.FungiForum.com
|
jccc
just aotherhuman
Registered: 12/01/06
Posts: 1,162
Last seen: 3 months, 8 days
|
Re: Best way to make an LC [Re: Cryogenicz]
#6599062 - 02/22/07 06:12 PM (5 years, 3 months ago) |
|
|
Dont make one!!! Just knock up some grains and then inoculate more grains with colonized grains...because im going through hell with my lcs trying to seperate bacteria in the lc that i cant see from good mycelium..with lcs u only have one chance and if u screww up it messes everything up!
|
mattymonkey
Feel Like aStranger...
Registered: 11/07/04
Posts: 973
Last seen: 1 year, 1 month
|
Re: Best way to make an LC [Re: jccc]
#6599093 - 02/22/07 06:16 PM (5 years, 3 months ago) |
|
|
LC is great.. just be sterile, and test them on a *single* jar before you knock up a ton.
--------------------
"listening for the secret.. searching for the sound.."
|
RogerRabbit
Bans for Pleasure
Registered: 03/26/03
Posts: 36,974
Loc: USA Mountain Northwest
Last seen: 12 hours, 29 minutes
|
Re: Best way to make an LC [Re: Cryogenicz]
#6599605 - 02/22/07 08:15 PM (5 years, 3 months ago) |
|
|
Quote:
Cryogenicz said:
What would you consider an intermediate microscope, which could view bacteria?
-Graham
I wouldn't go with less than one of these http://www.microscopesusa.com/Observer_III.html If you can afford it, look into the lab microscopes on that page. I have a trinocular M2 lab scope that is pretty good. To get better, you'd really have to move up to an olympus or similar that would cost four times as much.
RR
--------------------
www.mushroomvideos.com
semper in excretia sumus solim profundum variat
|
FreeSporePrints
Mycelium Jedi
Registered: 03/06/05
Posts: 1,130
Loc: Rome, ITALY
Last seen: 6 days, 9 hours
|
Re: Best way to make an LC [Re: RogerRabbit]
#6600721 - 02/23/07 01:22 AM (5 years, 3 months ago) |
|
|
Hello Roger, my stereoscope has got the light near the lens but also under that cicular glass (see the pic).
Bacteria has got always a "pencil" shape as you said?
Do you put directly the petri under it or put a tissue of mycelium in solution?
Thanks!
Fabio
--------------------
|
fyouseeku
Chemist
Registered: 05/25/06
Posts: 79
|
Re: Best way to make an LC [Re: RogerRabbit]
#6600991 - 02/23/07 04:40 AM (5 years, 3 months ago) |
|
|
Wow and I thought it was going to be a cake walk... I will just stick to the basic agar to grain then g2g it for a while.....
I know in another thread (not gourmet) RR you mentioned that you use to let your rye grain colonize about 80 - 100% then shoot distilled water into the grain, shake then remove water and that would dislodge enough mycelium to make a syringe.
Would this work?
|
chrisscole
Mr
Registered: 10/08/06
Posts: 102
Loc: Oxon, England
Last seen: 4 years, 9 months
|
Re: Best way to make an LC [Re: fyouseeku]
#6601362 - 02/23/07 08:38 AM (5 years, 3 months ago) |
|
|
there are easier ways. check out the FAQ pages for some shortcuts or search for threads on Liquid Cultures.
I managed to use a syringe I bought from somewhere else to inoculate a jar of malt extract and made another syringe from that - works a treat, and there wasn't any contam in the grain, so I'm thinking it was reasonably clean. Havn't had success with honey water though.
Planning on making a homemade stirrer at the moment, just got to get a computer fan and i'll have everything i need.
Chris
|
RogerRabbit
Bans for Pleasure
Registered: 03/26/03
Posts: 36,974
Loc: USA Mountain Northwest
Last seen: 12 hours, 29 minutes
|
Re: Best way to make an LC [Re: fyouseeku]
#6601378 - 02/23/07 08:45 AM (5 years, 3 months ago) |
|
|
Let it colonize 100% before you do that though. That's the fastest way to get contaminant free liquid mycelium syringes that I've ever found.
Fabio, to test your mycelium for bacteria, take a clean microscope slide and put one drop of distilled water in the center of it. Using a sterile tool or syringe needle, scrape a tiny amount of mycelium and swirl it into the drop of water on the slide. Cover it with a coverslip, which flattens everything out so it's all in focus. Use a light microscope for viewing at 100X to 400X. The picture below is what you'll be looking for. You can see an individual strand of dikaryotic mycelium. You can see the cell walls between individual cells, and the two nuclei in each cell(a couple of the cells have three). The bacterial rods if present would be roughly the same length as the cells of mycelium, but they'd be individual cells, not linked together. I have a video of bacteria on a slide, but didn't find any still pictures yet. I'll look some more.
RR
--------------------
www.mushroomvideos.com
semper in excretia sumus solim profundum variat
|
chrisscole
Mr
Registered: 10/08/06
Posts: 102
Loc: Oxon, England
Last seen: 4 years, 9 months
|
Re: Best way to make an LC [Re: RogerRabbit]
#6601634 - 02/23/07 10:08 AM (5 years, 3 months ago) |
|
|
this thread is becoming quite interesting...
so how do you tell the difference between a monokaryotic and a dikaryotic strand of mycelium on the microscope?
what did you mean by the 100% thing? let the LC culture, or the syringe, colonize 100%? before you stir it?
Chris
|
FreeSporePrints
Mycelium Jedi
Registered: 03/06/05
Posts: 1,130
Loc: Rome, ITALY
Last seen: 6 days, 9 hours
|
Re: Best way to make an LC [Re: chrisscole]
#6601841 - 02/23/07 11:02 AM (5 years, 3 months ago) |
|
|
Hi Roger, really thank you for your tips, when you'll find if you want you'll upload here.
chris: i'm supposing Roger was answering to fyouseeku, about colonizing 100% the substrate then insert distilled water to make a liquid culture syringe.
A dykariotic mycelium means that a single cell has got 2 nuclei (as you see in the pic) and a full complement of chromosomes, monokaryotic one has got a single nucleum.
--------------------
|
|
Previous
Index
Next
Threaded
Edit 
Reply 
