noob troubles with invitro

Sky Rayven · 09-22-06, 16:10

Hi Fellow Topiates. I made my jars last night. shot them up this morning.

I had major problems with the syringe. My motor controll isn't the greatest. I went through 10 CC for 3 jars. I shook the syringe to break up the spores. And then when I tried to get just a quarter of a CC into each of the four holes It was impossible and when I took my syringe back out of the holes there was spore solution leaking out of the syringe needle and I didn't really think that I was pushing on the plunger that hard. I was like WTF and God Dang it.

What advice could be given for this issue! I don't even know how much solution went into the jars and how much didn't.

Is this going to be a problem for the colonization process?

Please Help!!

Thanx Guys

max · 09-22-06, 16:26

Hopefully the extra solution is pooling in one place. If it is spread around nicely then you should be ok, just less jars than normal.

dial8 · 09-22-06, 16:37

I have to say, I love the "god dang it" part.

Well you should be ok...the worst that could happen is you have some jars that don't do well but I bet you have some that do.

Sky Rayven · 09-22-06, 16:47

Quote:

Originally Posted by captainmax

Hopefully the extra solution is pooling in one place. If it is spread around nicely then you should be ok, just less jars than normal.

Maxter...What do you supose was going on. Is the substrate getting stuck in the tip of the needle? There where no clumps in the syringe. Some times I would be pushing the plunger and It wouldn't move.

If the solution is pooling I don't see it. Should I kinda gently sortof rotate an tilt the jars a little to move the solution around?

Thanx Very much For your Help!!!!!

max · 09-22-06, 16:55

I've heard some people roll thier jars to spread it around a bit.

teknihil · 09-22-06, 17:01

Don't know if this will help you, but a similar thing happened to my FOAF... he thinks what happened was when he pushed the syringe in the hole, some of the substrate blocked the needle hole and prevented the liquid from coming out. So, he pushed the syringe all the way in, then pulled it out a centimeter or so. When he pushed the plunger, he let it sit there for a couple secs to make sure all the liquid came out of the needle. When he pulled it out, he made sure to not touch the plunger.

Outer · 09-22-06, 17:08

Sky the samething happened to me.After i used my syringe up i filled it up with water and practiced shooting quater cc at a time to get the feel of it.The stuck part happened too but i got lucky and it happened on the 7th jar but my spores shoot all over my glove box.Anyone know of a safe way to get your syringe unclogged?

Sky Rayven · 09-22-06, 17:09

Quote:

Originally Posted by dial8

I have to say, I love the "god dang it" part.

Well you should be ok...the worst that could happen is you have some jars that don't do well but I bet you have some that do.

hehe Glad you find it funny...

I have a sneaking suspicion that the substrate was pluging the needle a bit and then as I removed the needle it became un pluged releasing the solution throughout the middle parts of the sub. But hell I dunno. Some times the needle seemed a bit bent by about 10°.

Kay Serra, Whatever will be will be.

I just wish I knew a better way to get control on this.

Thanx!!

tbonus · 09-22-06, 17:18

Hopefully the jars will collonise REAL fast.Good luck

Sky Rayven · 09-22-06, 17:26

Quote:

Originally Posted by teknihil

Don't know if this will help you, but a similar thing happened to my FOAF... he thinks what happened was when he pushed the syringe in the hole, some of the substrate blocked the needle hole and prevented the liquid from coming out. So, he pushed the syringe all the way in, then pulled it out a centimeter or so. When he pushed the plunger, he let it sit there for a couple secs to make sure all the liquid came out of the needle. When he pulled it out, he made sure to not touch the plunger.

Makes tons of sense!! Will try this next time.

Sky Rayven · 09-22-06, 17:29

Quote:

Originally Posted by Outer

Sky the samething happened to me.After i used my syringe up i filled it up with water and practiced shooting quater cc at a time to get the feel of it.The stuck part happened too but i got lucky and it happened on the 7th jar but my spores shoot all over my glove box.Anyone know of a safe way to get your syringe unclogged?

What you did is probably the safest. Although wastful It did unclog and you didn't have to mess with the apparatus. But then what do I know!!

Pedestrian · 09-22-06, 18:10

One way to inject that works well for foaf is to insert the needle about 1/2 way into the jar, then pull it back about a 1/4 inch, then proceed to inject your desired ammount of solution. With this method foaf has not had any more clogging problems.

Cheers

-Ped

Lazlo · 09-22-06, 18:13

Yep, that's the best way to do it Pedestrian.

Your jars should be fine. Texas Bob says he shoots 5mills into a brf cake that contains 60mills of water and his fly without problems. I like to use 2mills per jar myself.

max · 09-22-06, 18:47

The best way is to insert the needle only a small bit, just past the verm layer. Then, don't squirt the solution so it shoots to the bottom but just let it dribble down so the spores stick to the substrate all the way down. Then give the jar a bit of a shake so the innoculation tunnels left by the needle can get plugged by some verm.

explorerowid · 09-22-06, 19:23

If there was spore solution leaking when you pulled the needle
out it could cause higher risk for contams. keep a close eye on them .
good luck
E.E.

Sky Rayven · 09-22-06, 19:57

Quote:

Originally Posted by explorerowid

If there was spore solution leaking when you pulled the needle
out it could cause higher risk for contams. keep a close eye on them .
good luck
E.E.

Point taken...But why would that be? Would you care to explain why that might cause higher risk for contaims. Please

Thanx explorerowid

Thanks to All

oibchip · 09-22-06, 20:01

Hey Sky, Are you getting any air in the Syringe, if so it needs to be expelled prior to the next injection. That will make it easier to shoot the correct amount.

Just my 3 cents

Sky Rayven · 09-22-06, 20:26

Quote:

Originally Posted by oibchip

Hey Sky, Are you getting any air in the Syringe, if so it needs to be expelled prior to the next injection. That will make it easier to shoot the correct amount.

Just my 3 cents

Yes...Air was in the syringe...as it should be for shaking. I did not expell that air b4 starting.

As Carlos Mencia would say..."DeeDeeDee"

Hippie3 · 09-22-06, 20:51

Quote:

Originally Posted by Pedestrian

One way to inject that works well for foaf is to insert the needle about 1/2 way into the jar, then pull it back about a 1/4 inch, then proceed to inject your desired ammount of solution. With this method foaf has not had any more clogging problems.

Cheers

-Ped

agreed
and of course
practice makes for perfection

oibchip · 09-22-06, 20:51

Expell the air B4 injecting and your injections will be much smoother

Hippie3 · 09-22-06, 20:51

Quote:

Originally Posted by Sky Rayven

Point taken...But why would that be? Would you care to explain why that might cause higher risk for contaims. Please

Thanx explorerowid

Thanks to All

because
if water can get out
air might be able to get in...

Elf Salvation · 09-22-06, 20:52

When i inocculate i stab the needle to a good depth and pull it back a bit, the solution should flow nicely.

Air in the syringe will cause pressure and expel water when ya take the needle out of jar, liquids dont pressurize that easily but gasses do.

Good luck

Sky Rayven · 09-22-06, 21:06

Quote:

Originally Posted by Hippie3

because
if water can get out
air might be able to get in...

and you too Elf Salvation

Much Appriciated

Would it be safe then to say a precautionary mesure for contams would be to rubberband a filter to the top of the jars using kyvek...I think that is how it is pronounced...the stuff you can get at the post office?

Elf Salvation · 09-22-06, 21:38

I use tyvek on every jar even the few pf jars i have done had tyvek and one of the twenty grew a white mycelia like cotton that was a black mold. Thoes pf inoculations where done in open air replacing tyvek ect.

Elf Salvation · 09-22-06, 21:40

Free postal tyvek envelope http://shop.usps.com/webapp/wcs/stor...1603&langId=-1

teknihil · 09-22-06, 21:43

May seem like simplistic stuff, but I learned a lot in this thread that I haven't seen in any teks. Probably out there somewhere I but never saw it. Thanks.

Sky Rayven · 09-22-06, 22:04

Quote:

Originally Posted by Elf Salvation

Free postal tyvek envelope http://shop.usps.com/webapp/wcs/stores/servlet/ProductDisplay?catalogId=10152&storeId=10001&categ oryId=13353&productId=11603&langId=-1

I just ordered me some Tyvek!! I hate going to the Post Office.

Free Delivery Free Product!!

Super Nice

Thanx Elf

BuckarooBanzai · 09-22-06, 22:12

When my FOAF inoculates, he goes in slowly at a steep angle, so the needle is actually touching the glass. Then he rotates the needle, until the hole is visible through the side of the glass (you can see if it is plugged). Then solution is added slowly, so it spreads against the glass and drips down the side. .3mL to .6mL is usually enough to drip all the way to the bottom of the jar. He stops when the liquid hits the bottom of the jar.

And yeah, practicing this with water and dummy jars is a very good idea. Especially if you have never handled needles before.

But don't worry, you'll get the hang of it!!!

Sky Rayven · 09-22-06, 23:12

I have handled needles a lot. Thatz the wierd thing.

Practice will make perfect.

I am not givin' up.

Sky Rayven · 09-25-06, 16:04

Ok 3 days later...this is too soon i suppose. but no contaims that i can see. Also I may have spotted a little bit of white fuz on the side of one of the jars really hard to see.

Now...my jars are incubating at about 82°. I can't get my heater to stay at 80° but the two choices seem to be 78.5° or 82.2° roughly. Is warmer better than cooler?

Outer · 09-25-06, 16:39

Quote:

Originally Posted by Pedestrian

One way to inject that works well for foaf is to insert the needle about 1/2 way into the jar, then pull it back about a 1/4 inch, then proceed to inject your desired ammount of solution. With this method foaf has not had any more clogging problems.

Cheers

-Ped

Quote:

Originally Posted by captainmax

Then give the jar a bit of a shake so the innoculation tunnels left by the needle can get plugged by some verm.

nice tips thanks

BuckarooBanzai · 09-25-06, 21:53

I would say 82.2 is better. The faster you can get the jars to grow, the better the chance your mycelia will take over before any contaminant can.

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09-22-06, 16:10	#1 (permalink)
Sky Rayven Mycotopiate Join Date: Aug 2006 Posts: 652	noob troubles with invitro Hi Fellow Topiates. I made my jars last night. shot them up this morning. I had major problems with the syringe. My motor controll isn't the greatest. I went through 10 CC for 3 jars. I shook the syringe to break up the spores. And then when I tried to get just a quarter of a CC into each of the four holes It was impossible and when I took my syringe back out of the holes there was spore solution leaking out of the syringe needle and I didn't really think that I was pushing on the plunger that hard. I was like WTF and God Dang it. What advice could be given for this issue! I don't even know how much solution went into the jars and how much didn't. Is this going to be a problem for the colonization process? Please Help!! Thanx Guys __________________ The Goddess is alive and she ate my mushrooms! ;)

09-22-06, 17:18	#9 (permalink)
tbonus Mycotopiate Join Date: May 2006 Posts: 509	Hopefully the jars will collonise REAL fast.Good luck __________________ TWO WRONGS DONT MAKE A RIGHT, BUT THREE RIGHTS MAKE A LEFT!

09-22-06, 18:10	#12 (permalink)
Pedestrian Dreamspace Transient Join Date: Feb 2006 Posts: 563	One way to inject that works well for foaf is to insert the needle about 1/2 way into the jar, then pull it back about a 1/4 inch, then proceed to inject your desired ammount of solution. With this method foaf has not had any more clogging problems. Cheers -Ped __________________ Intuition is superior to logic.

09-22-06, 18:13	#13 (permalink)
Lazlo old hand Join Date: Mar 1970 Posts: 7,059	Yep, that's the best way to do it Pedestrian. Your jars should be fine. Texas Bob says he shoots 5mills into a brf cake that contains 60mills of water and his fly without problems. I like to use 2mills per jar myself. __________________ How can you have any pudding, if you don't eat your MEAT?

09-22-06, 16:26	#2 (permalink)
max canadian Join Date: Apr 1971 Posts: 854	Hopefully the extra solution is pooling in one place. If it is spread around nicely then you should be ok, just less jars than normal.

09-22-06, 16:37	#3 (permalink)
dial8 Un Undead ***er8tor Join Date: Jun 1976 Posts: 3,185	I have to say, I love the "god dang it" part. Well you should be ok...the worst that could happen is you have some jars that don't do well but I bet you have some that do.

09-22-06, 16:55	#5 (permalink)
max canadian Join Date: Apr 1971 Posts: 854	I've heard some people roll thier jars to spread it around a bit.

09-22-06, 17:01	#6 (permalink)
teknihil Mycophiliac Join Date: Aug 2006 Posts: 9	Don't know if this will help you, but a similar thing happened to my FOAF... he thinks what happened was when he pushed the syringe in the hole, some of the substrate blocked the needle hole and prevented the liquid from coming out. So, he pushed the syringe all the way in, then pulled it out a centimeter or so. When he pushed the plunger, he let it sit there for a couple secs to make sure all the liquid came out of the needle. When he pulled it out, he made sure to not touch the plunger.

09-22-06, 17:08	#7 (permalink)
Outer Mycophage Join Date: Jul 2006 Posts: 177	Sky the samething happened to me.After i used my syringe up i filled it up with water and practiced shooting quater cc at a time to get the feel of it.The stuck part happened too but i got lucky and it happened on the 7th jar but my spores shoot all over my glove box.Anyone know of a safe way to get your syringe unclogged?

09-22-06, 18:47	#14 (permalink)
max canadian Join Date: Apr 1971 Posts: 854	The best way is to insert the needle only a small bit, just past the verm layer. Then, don't squirt the solution so it shoots to the bottom but just let it dribble down so the spores stick to the substrate all the way down. Then give the jar a bit of a shake so the innoculation tunnels left by the needle can get plugged by some verm.

09-22-06, 19:23	#15 (permalink)
explorerowid Mycophage Join Date: Jun 2006 Posts: 102	If there was spore solution leaking when you pulled the needle out it could cause higher risk for contams. keep a close eye on them . good luck E.E.

09-22-06, 20:01	#17 (permalink)
oibchip Space Lord Modulator Join Date: May 2006 Posts: 5,065	Hey Sky, Are you getting any air in the Syringe, if so it needs to be expelled prior to the next injection. That will make it easier to shoot the correct amount. Just my 3 cents

09-22-06, 20:51	#20 (permalink)
oibchip Space Lord Modulator Join Date: May 2006 Posts: 5,065	Expell the air B4 injecting and your injections will be much smoother

09-22-06, 20:52	#22 (permalink)
Elf Salvation Pann Man Join Date: Nov 2005 Posts: 590	When i inocculate i stab the needle to a good depth and pull it back a bit, the solution should flow nicely. Air in the syringe will cause pressure and expel water when ya take the needle out of jar, liquids dont pressurize that easily but gasses do. Good luck

09-22-06, 21:38	#24 (permalink)
Elf Salvation Pann Man Join Date: Nov 2005 Posts: 590	I use tyvek on every jar even the few pf jars i have done had tyvek and one of the twenty grew a white mycelia like cotton that was a black mold. Thoes pf inoculations where done in open air replacing tyvek ect.

09-22-06, 21:40	#25 (permalink)
Elf Salvation Pann Man Join Date: Nov 2005 Posts: 590	Free postal tyvek envelope http://shop.usps.com/webapp/wcs/stor...1603&langId=-1

09-22-06, 21:43	#26 (permalink)
teknihil Mycophiliac Join Date: Aug 2006 Posts: 9	May seem like simplistic stuff, but I learned a lot in this thread that I haven't seen in any teks. Probably out there somewhere I but never saw it. Thanks.

09-22-06, 22:12	#28 (permalink)
BuckarooBanzai Prone to ranting... Join Date: Oct 2005 Posts: 4,243	When my FOAF inoculates, he goes in slowly at a steep angle, so the needle is actually touching the glass. Then he rotates the needle, until the hole is visible through the side of the glass (you can see if it is plugged). Then solution is added slowly, so it spreads against the glass and drips down the side. .3mL to .6mL is usually enough to drip all the way to the bottom of the jar. He stops when the liquid hits the bottom of the jar. And yeah, practicing this with water and dummy jars is a very good idea. Especially if you have never handled needles before. But don't worry, you'll get the hang of it!!! __________________ Banzai Institute for Higher Education (a collection of growing Teks & threads)

09-22-06, 23:12	#29 (permalink)
Sky Rayven Mycotopiate Join Date: Aug 2006 Posts: 652	I have handled needles a lot. Thatz the wierd thing. Practice will make perfect. I am not givin' up. __________________ The Goddess is alive and she ate my mushrooms! ;)

09-25-06, 16:04	#30 (permalink)
Sky Rayven Mycotopiate Join Date: Aug 2006 Posts: 652	Incubator Temp Ok 3 days later...this is too soon i suppose. but no contaims that i can see. Also I may have spotted a little bit of white fuz on the side of one of the jars really hard to see. Now...my jars are incubating at about 82°. I can't get my heater to stay at 80° but the two choices seem to be 78.5° or 82.2° roughly. Is warmer better than cooler? __________________ The Goddess is alive and she ate my mushrooms! ;)

09-25-06, 21:53	#32 (permalink)
BuckarooBanzai Prone to ranting... Join Date: Oct 2005 Posts: 4,243	I would say 82.2 is better. The faster you can get the jars to grow, the better the chance your mycelia will take over before any contaminant can. __________________ Banzai Institute for Higher Education (a collection of growing Teks & threads)