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OfflineEmperor Reishi
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Working with spore prints and inoculation rings...
    #9369034 - 12/04/08 11:56 PM (12 years, 2 months ago)

So, i haven't done much work with spore prints yet. I fashioned a home-made inoculation ring, but when I tried to scrape my spores of the prints (my pearl oysters just powdered up on the print paper and didn't really transfer to my ring and my golden oysters seemed to go on the ring a bit, but i could hardly see them)  Now i'll see in the next few days how the transfer went, but does anybody have any suggestions or input?


--------------------
EmperoR ReishI

Looking for:    Live Cultures: Maitake[sclerotia forming], Wood Ear[Auricularia Polytricha],            Prints: **Psilocybe mexicana,                                Reishi [other than red], Bioluminessent                                                                          Cubes & Pan Cyans

Available for trade:    Live cultures: Reishi[ganoderma lucidum], Shiitake[lentinula edodes], Chicken of the woods[Laetiporus                                            sulphureus], Enoki [Flammulina velutipes], Oyster [Pleurotus ostreatus], Lion's Mane [Hericium                                                            erinaceus]


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Offlinesolumvita
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Re: Working with spore prints and inoculation rings... [Re: Emperor Reishi]
    #9369268 - 12/05/08 12:40 AM (12 years, 2 months ago)

A good way to get the loop sticky is to flame the loop till it is red hot then cool it in the agar plate/tube you are going to transfer the spores into, that will coat the loop and cool it simultaneously. 

Then you take a scraping of the spore print and transfer to your tube.


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Offlinedenger
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Re: Working with spore prints and inoculation rings... [Re: solumvita]
    #9369633 - 12/05/08 01:37 AM (12 years, 2 months ago)

What did you make your loop out of?


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OfflineEmperor Reishi
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Re: Working with spore prints and inoculation rings... [Re: denger]
    #9370609 - 12/05/08 07:27 AM (12 years, 2 months ago)

solumvita:
Thanks for the tip. I'll try that on my next batch.

denger:
:wink: Well, it may sound silly, but it seemed like a good idea at the time, and I think it will work with solumvita's tip. I do a lot of remodeling around my house and electric work. I used about an 18" piece of 14 guage electrical wire. I striped the insulating plastic off about 1/4"-3/8" in the center. I wrapped the center exposed metal around a chopstip to fashion the loop, and then twisted the ends together to hold the loop in place. and then used a bit of electrical tape to hold the handle together. I can take some pictures and form an in depth TEk if there is enough interest :tongue: heheheh, no but seriously if you wanna see a picture I can upload one just for shits and giggles.


--------------------
EmperoR ReishI

Looking for:    Live Cultures: Maitake[sclerotia forming], Wood Ear[Auricularia Polytricha],            Prints: **Psilocybe mexicana,                                Reishi [other than red], Bioluminessent                                                                          Cubes & Pan Cyans

Available for trade:    Live cultures: Reishi[ganoderma lucidum], Shiitake[lentinula edodes], Chicken of the woods[Laetiporus                                            sulphureus], Enoki [Flammulina velutipes], Oyster [Pleurotus ostreatus], Lion's Mane [Hericium                                                            erinaceus]


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Offlinedenger
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Re: Working with spore prints and inoculation rings... [Re: Emperor Reishi]
    #9370798 - 12/05/08 09:28 AM (12 years, 2 months ago)

Quote:

Emperor Reishi said:
I used about an 18" piece of 14 guage electrical wire.




This is a pretty thick wire. It is probably copper too. This means you will have a hard time heating it to the recommended cherry-red colour. I would recommend using thinner wire. You can get thin stainless steel wire at a hardware store.


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OfflineEmperor Reishi
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Re: Working with spore prints and inoculation rings... [Re: denger]
    #9370872 - 12/05/08 09:59 AM (12 years, 2 months ago)

Yeah, I knew copper wasn't the best choice (I was thinking because it oxidizes so easily), but i thought I'd give it a shot to use for now... 14 guage is pretty thin.... probably about 1/16" in diameter. Should it be much thinner (i'm really not sure)

Copper conducts heat really well though, that's why really nice cookware has the copper plate in the bottom... but i've been having trouble heating my tools cherry. My alcohol lamp keeps snuffing out on me, I need to get a better one, or experiment to get this one to work better. I've been mostly relying on heating pretty hot and also following up cleaning with alcohol.

I've also been unsing a stupid x-acto blade. I'm going to buy a nice scalpel off e-bay soon. I've seen a variety on ebay that stamets recommends and it's only like $7 with 100 blades i think as opposed to like $15 on his site with only several blades i think.


--------------------
EmperoR ReishI

Looking for:    Live Cultures: Maitake[sclerotia forming], Wood Ear[Auricularia Polytricha],            Prints: **Psilocybe mexicana,                                Reishi [other than red], Bioluminessent                                                                          Cubes & Pan Cyans

Available for trade:    Live cultures: Reishi[ganoderma lucidum], Shiitake[lentinula edodes], Chicken of the woods[Laetiporus                                            sulphureus], Enoki [Flammulina velutipes], Oyster [Pleurotus ostreatus], Lion's Mane [Hericium                                                            erinaceus]


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OfflineParesthesia
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Re: Working with spore prints and inoculation rings... [Re: Emperor Reishi]
    #9371109 - 12/05/08 11:18 AM (12 years, 2 months ago)

Making loops is a lot faster if you use mechanical assistance.  I've made tons of twisted wire for a friend who makes jewelry using a nail and an electric drill.  This will also let you use a stiffer wire.  I also wouldn't secure it using tape, as it can be difficult to keep clean.  Good idea, though.  It could save a few bucks!

I also don't see why an x-acto blade wouldn't work.  You should autoclave the knife when you PC things, though.  Stuff could get into the blade receptacle.  Maybe.  I don't know. :smile:


--------------------
"We shall not cease from exploration, and the end of all our exploring will be to arrive where we started and know the place for the first time."

- T. S. Eliot


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OfflineEmperor Reishi
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Re: Working with spore prints and inoculation rings... [Re: Paresthesia]
    #9371858 - 12/05/08 01:40 PM (12 years, 2 months ago)

Quote:

Paresthesia said:
I also don't see why an x-acto blade wouldn't work.  You should autoclave the knife when you PC things, though.  Stuff could get into the blade receptacle.  Maybe.  I don't know. :smile:




I dunno, I was hoping the scalpel would be sharper because FME the x-actos don't seem to cut through certain things very well... they seem dull. For instance when i was trasfering a piece of very well colonized media it was like trying to cut a piece of rubber with a butter knife...


--------------------
EmperoR ReishI

Looking for:    Live Cultures: Maitake[sclerotia forming], Wood Ear[Auricularia Polytricha],            Prints: **Psilocybe mexicana,                                Reishi [other than red], Bioluminessent                                                                          Cubes & Pan Cyans

Available for trade:    Live cultures: Reishi[ganoderma lucidum], Shiitake[lentinula edodes], Chicken of the woods[Laetiporus                                            sulphureus], Enoki [Flammulina velutipes], Oyster [Pleurotus ostreatus], Lion's Mane [Hericium                                                            erinaceus]


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OfflineParesthesia
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Re: Working with spore prints and inoculation rings... [Re: Emperor Reishi]
    #9371926 - 12/05/08 01:51 PM (12 years, 2 months ago)

It's like that with a scalpel, too.  It takes a fair amount of pressure to break through a well colonized piece of king oyster mycelium, for instance.

The sharpest thing to use would be an obsidian glass scalpel blade!  Maybe a ceramic blade comes close.  You'd have to be careful not to drop it, and you'd still need to use a lot of pressure to cut through tough mycelium.


--------------------
"We shall not cease from exploration, and the end of all our exploring will be to arrive where we started and know the place for the first time."

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Offlinespazn420
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Re: Working with spore prints and inoculation rings... [Re: Paresthesia]
    #9372095 - 12/05/08 02:20 PM (12 years, 2 months ago)

here's my inoculation loop. It is a safety pin and a pencil. Work perfect.



--------------------


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OfflineEmperor Reishi
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Re: Working with spore prints and inoculation rings... [Re: Paresthesia]
    #9372125 - 12/05/08 02:26 PM (12 years, 2 months ago)

Gotcha, thanks for the tip. I guess I'll just stick with my xacto for now then :smile:, I do want to find a good deal on a magnetic stir plate though :blush:

Also though, do you guys have the issue of your scalpel and/or loops turning black when flame sterilizing, how do you avoid this. Am I just holding too close into the flame?


--------------------
EmperoR ReishI

Looking for:    Live Cultures: Maitake[sclerotia forming], Wood Ear[Auricularia Polytricha],            Prints: **Psilocybe mexicana,                                Reishi [other than red], Bioluminessent                                                                          Cubes & Pan Cyans

Available for trade:    Live cultures: Reishi[ganoderma lucidum], Shiitake[lentinula edodes], Chicken of the woods[Laetiporus                                            sulphureus], Enoki [Flammulina velutipes], Oyster [Pleurotus ostreatus], Lion's Mane [Hericium                                                            erinaceus]


Edited by Emperor Reishi (12/05/08 02:39 PM)


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Offlinedenger
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Re: Working with spore prints and inoculation rings... [Re: Emperor Reishi]
    #9372288 - 12/05/08 02:53 PM (12 years, 2 months ago)

It is normal for metal tools to oxidise and turn black after proper flaming. The blade on my scalpel degrades gradually, and I have to replace it every 15-20 sessions. My loop is made from nickel wire, and it does not seem to degrade significantly, I've been using it for several years now.
I looked around and the loop wire seems to be about the same thickness as the conductor of the telephone/network wire (the solid conductor type used for permanent wiring in walls). You can get a foot of solid conductor network cable at home depot for a buck give or take. It will still be copper, but more flexible and easier to flame sterilize. Same place should have even better choice - stainless steel wire in thin gauge (20 or so).


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OfflineEmperor Reishi
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Re: Working with spore prints and inoculation rings... [Re: denger]
    #9377402 - 12/06/08 07:29 AM (12 years, 2 months ago)

yeah, with an xacto blade it turns black instaltly pretty much. The very first flame sterilization and my blade gets all black, and the black (carbon?) rubs off on my work, be it spore prints of agar... ect... so I end up flaming and then alcohol swabbing.... I was thinking the scalpel would have higher quality metal blades being there function calls for that, and was hoping they would degrade much slower...

Thanks for the tip on the 'noc loop, I'll probably make a new one this weekend. I need to go to Lowes/Home Depot anyway. I need some tubing to make air ports on my LCs.

Later guys


--------------------
EmperoR ReishI

Looking for:    Live Cultures: Maitake[sclerotia forming], Wood Ear[Auricularia Polytricha],            Prints: **Psilocybe mexicana,                                Reishi [other than red], Bioluminessent                                                                          Cubes & Pan Cyans

Available for trade:    Live cultures: Reishi[ganoderma lucidum], Shiitake[lentinula edodes], Chicken of the woods[Laetiporus                                            sulphureus], Enoki [Flammulina velutipes], Oyster [Pleurotus ostreatus], Lion's Mane [Hericium                                                            erinaceus]


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Offlinedenger
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Re: Working with spore prints and inoculation rings... [Re: Emperor Reishi]
    #9377548 - 12/06/08 09:08 AM (12 years, 2 months ago)

If you get carbon deposit on your blade (soot) it means you are flaming it too low in the flame. The hottest part of the flame is right above the tip of the low-light zone, and you should aim your tools above this part:
http://www.buzzlites.ca/flame.html


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Re: Working with spore prints and inoculation rings... [Re: denger]
    #9379182 - 12/06/08 04:02 PM (12 years, 2 months ago)

Another way to get the black off the blade is once it is really red, i dip into alcohol and it "boils" all the black off.


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Offlinedenger
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Re: Working with spore prints and inoculation rings... [Re: Emperor Reishi]
    #9379337 - 12/06/08 04:24 PM (12 years, 2 months ago)

Quote:

Emperor Reishi said:
I do want to find a good deal on a magnetic stir plate though




You can make your own. I made one for under $15:
http://www.shroomery.org/forums/showflat.php/Cat/0/Number/9337138/an/0/page/0


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Re: Working with spore prints and inoculation rings... [Re: solumvita]
    #9381046 - 12/06/08 08:59 PM (12 years, 2 months ago)

Quote:

solumvita said:
Another way to get the black off the blade is once it is really red, i dip into alcohol and it "boils" all the black off.




This sounds a little dangerous. Apparently the metal is not hot enough to ignite the alcohol (assuming you still have your eyebrows :wink:), but having open pools of alcohol and a flame in the same proximity is asking for trouble.

-DD


--------------------
" learning how to cook will save you money, usually make you healthier, and occasionally get you laid. "
-Shags420

"Most of the people who ask the question 'Do any psilocybin mushrooms grow around here?' would rather change their way of looking at reality rather than face the difficult and discouraging task of transforming reality itself"
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Re: Working with spore prints and inoculation rings... [Re: didjin_d]
    #9383636 - 12/07/08 07:19 AM (12 years, 2 months ago)

Well so far i have been doing it for years and i still have all my eyebrows, of course people that sue companies for hot coffee burning their lips probably will lose their eyebrows.  So I will have to add a disclaimer, use the previous suggestion at your own risk.  If you manage to lose your eyebrows that's your problem.  :hehehe:


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OfflineEmperor Reishi
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Re: Working with spore prints and inoculation rings... [Re: solumvita]
    #9390298 - 12/08/08 07:41 AM (12 years, 2 months ago)

Thankx for the tips guys.

Denger: Yeah, I wasn't sure if I was holding it too close, but now that you say that it comes bake to me & i'm going to check out that thread on building the stir plate, I think it would really help. I have three species brewing right now, and it takes me about a week at least to get to a usable point...

solumvita: I'll probably keep a little dish with alcohol in my glovebox and try that, but I missed your last post with the disclaimer... <cough> I'm gonna sue your a$s <ehem>...


--------------------
EmperoR ReishI

Looking for:    Live Cultures: Maitake[sclerotia forming], Wood Ear[Auricularia Polytricha],            Prints: **Psilocybe mexicana,                                Reishi [other than red], Bioluminessent                                                                          Cubes & Pan Cyans

Available for trade:    Live cultures: Reishi[ganoderma lucidum], Shiitake[lentinula edodes], Chicken of the woods[Laetiporus                                            sulphureus], Enoki [Flammulina velutipes], Oyster [Pleurotus ostreatus], Lion's Mane [Hericium                                                            erinaceus]


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