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InvisibleStropharis
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Stro's Spore Salvage with Forced Hydration * 7
    #15383615 - 11/17/11 01:02 PM (9 years, 7 months ago)

Stro's Spore Salvage with Forced Hydration


This post was edited for an easier read as a TEK.;

Having problems with old spores?

A simple rubber band wrapped around a syringe will offer the needed PSI to force hydrate a spore by forcing moisture through the cell walls, also know as hyper hydration.

The following thread includes my initial idea and other links of posts and threads that gave me this idea. I hope it helps, it did for me.



ORIGINAL THREAD BELOW

I have spores about 7 years old that I am having trouble with. I remembered seeing this post a long time ago and it took me forever to find again. Then I remembered it was a reply in reference to the pressure chamber RR built during the Redboy experiments. Unfortunately during the thread there was no respons to the idea. From the looks of it, it is a very simple and clever idea. Can anyone punch holes in this idea? Anyone ever use this technique? This one might just need to be posted as a TEK somewhere.

Quote:

Auxin said:
I was pondering upon this when an idea struck me. Why not use syringes for the pressure? The difficulty might be capping them off. Pre-filled medical syringes come with a good looking cap on them that may be autoclavable so I did an experiment. A standard 10CC syringe has a 1.5 cm diameter plunger, thats a surface area of .273 in² (the rubber plunger tip has a slight taper, physics 101 was too long ago and I dont remember how thatd effect pressure, but assuming .273 works in the calculation...) I placed 2.1 pounds of steel on the plunger of a sealed syringe while it was supported vertically to produce a pressure of 7.7 PSI on the plunger top, I left it there for 5 minutes and not a single microdrop of water leaked through the syringe end cap. It should be just like pressurizing a jar with air and no worries about filtering air from a pump, something to think about :smile:






Edited by Stropharis (06/17/13 09:30 AM)


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InvisibleJavadog
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Re: Pressure Syringe Spore Salvage [Re: Stropharis]
    #15383719 - 11/17/11 01:32 PM (9 years, 7 months ago)

An excellent post Stropharis.

I am too new to have spores old enough to need special treatment,
but look forward to how this plays out.

Good luck,

JD


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InvisibleStropharis
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Re: Pressure Syringe Spore Salvage [Re: Javadog]
    #15383795 - 11/17/11 01:52 PM (9 years, 7 months ago)

Just found a calculator online.



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InvisibleJavadog
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Re: Pressure Syringe Spore Salvage [Re: Stropharis]
    #15383857 - 11/17/11 02:05 PM (9 years, 7 months ago)

Great link.  Thanks!


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InvisibleStropharis
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Re: Pressure Syringe Spore Salvage [Re: Javadog]
    #15384050 - 11/17/11 02:52 PM (9 years, 7 months ago)

The following is by no means accurate but as long as I am pressurizing it I am getting the job done. While trying to figure out how to prop 2lbs on top of the syring I came up with another idea. Rubberbands.


Like I said this isn't accurate but it gets the job done. I am holding just enough pressure to keep the band off the scale simulating the size of the glass with my hand. Even though there is only 6ml of solution, its still is a BD 10ml with an internal diameter of 0.561, approximately 2lbs of pressure gives me approximately 8psi

I tossed another rubber band for good measure, which in fact makes the measurements further off, lol. I would say it is somewhere between 5-10 psi though.


Edited by Stropharis (11/17/11 02:56 PM)


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OfflineSKrink
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Re: Pressure Syringe Spore Salvage [Re: Stropharis] * 1
    #15384203 - 11/17/11 03:29 PM (9 years, 7 months ago)

How will pressurizing a syringe salvage old spores? This looks very interesting and I'd like to know more.


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... Oh mighty masticator, salivator, vocalizer, swallower, licker biter sucker brow-knitter looker blinker rubbernecker thumber prodder up-yours fingerer ringwearer nosepicker waver drinker armlifter bodybender hipswiveler kneer springer runner ZERO::::::::OOOOOOOOO:::::::: RUN!!!


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InvisibleStropharis
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Re: Pressure Syringe Spore Salvage [Re: SKrink]
    #15384236 - 11/17/11 03:37 PM (9 years, 7 months ago)

Quote:

SKrink said:
How will pressurizing a syringe salvage old spores? This looks very interesting and I'd like to know more.




It's forced hydration. The pressure forces moisture through the cell walls. I read about it somewhere and ran a search here to find it mentioned multiple times.


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InvisibleJavadog
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Re: Pressure Syringe Spore Salvage [Re: Stropharis]
    #15384308 - 11/17/11 03:51 PM (9 years, 7 months ago)

I love how you tricked the scale to tell you what you needed to know.

Let us know how it all shakes out.

Take care,

JD


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InvisibleStropharis
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Re: Pressure Syringe Spore Salvage [Re: Javadog]
    #15384334 - 11/17/11 03:56 PM (9 years, 7 months ago)

Quote:

Javadog said:
I love how you tricked the scale to tell you what you needed to know.

Let us know how it all shakes out.

Take care,

JD




Yea, I did my best not to actually, just lifting my finger ever so slightly off the scale it would land on 2lbs and any higher it would go below. As stated, not accurate but gets the job done of getting in the ballpark.


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InvisibleStropharis
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Re: Pressure Syringe Spore Salvage [Re: Stropharis]
    #15420727 - 11/25/11 05:05 AM (9 years, 6 months ago)

I ended up taking the syringes out of the glass and simply wrapping the rubber bands around the syringe.

There is one particular syringe that I have been working with for a while now. I have tried to start a culture on agar three different times with no success over the past month until now. I left the syringe pressurized for about five days and then started another dish, three day later we have germination. :rockon:


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Re: Pressure Syringe Spore Salvage [Re: Stropharis]
    #15421552 - 11/25/11 01:12 PM (9 years, 6 months ago)

Quote:

Stropharis said:
I ended up taking the syringes out of the glass and simply wrapping the rubber bands around the syringe.

There is one particular syringe that I have been working with for a while now. I have tried to start a culture on agar three different times with no success over the past month until now. I left the syringe pressurized for about five days and then started another dish, three day later we have germination. :rockon:




:hammertime:

Great work!


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Offlinek00laid
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Re: Pressure Syringe Spore Salvage [Re: Javadog]
    #15421569 - 11/25/11 01:16 PM (9 years, 6 months ago)

this thread rocks.

where does one go about finding these epic syringe caps.

the translucent blue ones that come from sponsors are not very water tight.

IMO/IME


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Invisiblehamloaf
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Re: Pressure Syringe Spore Salvage [Re: Javadog]
    #15421580 - 11/25/11 01:18 PM (9 years, 6 months ago)

Quote:

Javadog said:
Quote:

Stropharis said:
I ended up taking the syringes out of the glass and simply wrapping the rubber bands around the syringe.

There is one particular syringe that I have been working with for a while now. I have tried to start a culture on agar three different times with no success over the past month until now. I left the syringe pressurized for about five days and then started another dish, three day later we have germination. :rockon:




:hammertime:

Great work!



:imwithstupid:


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How I Get Stuff done. - My Reference Guide. - My Grows.


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InvisibleStropharis
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Re: Pressure Syringe Spore Salvage [Re: k00laid]
    #15421656 - 11/25/11 01:39 PM (9 years, 6 months ago)

Quote:

k00laid said:
this thread rocks.

where does one go about finding these epic syringe caps.

the translucent blue ones that come from sponsors are not very water tight.

IMO/IME




I am not entirely sure what caps come with the vendor syringes or where to get the ones I am using, I got them from a friend a long time ago. You would be surprised how little pressure is necessary to accomplish 2lbs on the syringe plunger. Fill a syringe with water, put the cap on, and see if you can wrap a rubber band around it a few times with out it leaking, now I am curious.


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Offlinek00laid
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Re: Pressure Syringe Spore Salvage [Re: Stropharis]
    #15421696 - 11/25/11 01:49 PM (9 years, 6 months ago)

i'll get back to this thread in a few days when im back home.

doing holiday stuffs at the moment.


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Re: Pressure Syringe Spore Salvage [Re: k00laid]
    #15421752 - 11/25/11 02:04 PM (9 years, 6 months ago)

I like it!:congrats:I wonder what else we could do along the same lines, a super cheap vacuum filter?


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Offlineblackout
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Re: Pressure Syringe Spore Salvage [Re: SomeGuy] * 1
    #15422159 - 11/25/11 03:44 PM (9 years, 6 months ago)

Very good. Another way to get over the balance issue would be to get a bit of styrofoam packaging, poke 3-4 syringe size holes in it about 4 inches apart, a grid shape for 4 and a triangular shape for 3, pop 3-4 syringes in the holes so they are standing upright with plungers in the air and you can balance a book or some other weights on the plungers.

Quote:

Here's the pressure chamber I built during the Redboy experiments to help hydrate old spores.  Glass jars are not designed to hold pressure, so I was afraid to ever go over 5 to 7 psi with it.
RR


A syringe will probably a lot more pressure. I expect the plunger stem would buckle first, it could easily be reinforced.


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OfflineSKrink
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Re: Pressure Syringe Spore Salvage [Re: Stropharis]
    #15422206 - 11/25/11 03:54 PM (9 years, 6 months ago)

Quote:

k00laid said:
this thread rocks.



:whathesaid:

I have a dessicated Orissa India cube print that I will try this on.
Never been able to get these spores to germinate on agar or grain, even with hydration, over multiple attempts.
Pressurized syringe here I come!

Quote:

Stropharis said:
I ended up taking the syringes out of the glass and simply wrapping the rubber bands around the syringe.




Can we get pics of the rubberbanded syringe w/o the jar? I'm just wondering how to tell I've got enough pressure (if the cap doesn't leak that is).

For example, if I start with liquid filled to the 10cc mark, should the plunger get pushed down noticeable and by how many units?

Oh yes and 5 :mushroom2: for you.


--------------------
:crazy2: SWEET POTATO HOME FRIES :crazy2:
:mushroom2:HOW TO USE A PENIS ENVY SPORE SWAB:mushroom2:

   

... Oh mighty masticator, salivator, vocalizer, swallower, licker biter sucker brow-knitter looker blinker rubbernecker thumber prodder up-yours fingerer ringwearer nosepicker waver drinker armlifter bodybender hipswiveler kneer springer runner ZERO::::::::OOOOOOOOO:::::::: RUN!!!


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Re: Pressure Syringe Spore Salvage [Re: blackout]
    #15422226 - 11/25/11 03:59 PM (9 years, 6 months ago)

This could essentially work but you will need to calculate the osmotic pressure to prevent the spores from lysing. This is the reason we use pressure sterilizers to destroy contamination.

Π = iMRT,
where

i is the dimensionless van 't Hoff factor
M is the molarity
R=0.0821 L atm K-1 mol-1 is the gas constant
T is the thermodynamic (absolute) temperature

You, or someone else, may have to find the pressure point at which spores are able to be fully hydrated without lysing.

Good luck, and I hope you are able to revive your spores.


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InvisibleStropharis
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Re: Pressure Syringe Spore Salvage [Re: XFINITY]
    #15422400 - 11/25/11 04:37 PM (9 years, 6 months ago)

Quote:

XFINITY said:
This could essentially work but you will need to calculate the osmotic pressure to prevent the spores from lysing. This is the reason we use pressure sterilizers to destroy contamination.

Π = iMRT,
where

i is the dimensionless van 't Hoff factor
M is the molarity
R=0.0821 L atm K-1 mol-1 is the gas constant
T is the thermodynamic (absolute) temperature

You, or someone else, may have to find the pressure point at which spores are able to be fully hydrated without lysing.

Good luck, and I hope you are able to revive your spores.




Lysing in relation to hyperhydration (forced hydration) is called cytolysis. It is caused by excessive osmosis and hydration towards the inside of a cell. The cell cannot withstand the osmotic pressure of the water inside, thus exploding.

At what PSI does this take place, I have no idea.


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