Sowing Seed Question

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zxyqu
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Sowing Seed Question

Post by zxyqu » Wed Aug 31, 2011 1:58 pm

Ok, so having done a bit of homework, I've figured out pretty much everything I would need once I have a seed pod, with one obvious question mark. Once you gotten your media jars and pod/seed sterilized, do you have to have a laminar flow hood to keep them sterile upon sowing? The area obviously needs to be quite sterile, but are there other options. I was thinking along the lines a fashioning a plexiglass/wood setup with gloves and a UV sterilization light. But honestly, I dont find a lot of comments on this step. People just go right from sterilizing seed/media to some sort of close up of a table and the sowing, with no mention of where/what they are in.

Thanks for any comments!
>> J <<
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TxRobNLA
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Re: Sowing Seed Question

Post by TxRobNLA » Wed Aug 31, 2011 2:49 pm

What you are describing is called a glove box, you should be able to do some searches for that online and find some plans and directions for using one.

At every stage you have to keep everything completely sterile. So you can never open a sterile flask to open air that has not been sterilized. There are mold and bacteria spores floating in all untreated air.

That said, in the old days, many orchid growers that were very successful at flasking did not have access to laminar flow hoods and they did all their labwork in a glove box. So it can definitely be done. You just have to be really aware of your environment and it will probably take some practice before you get it right.

In general you are going to want to sterilize everything in it with something like bleach. Even in my laminar flow hood, I have a towel on that has been soaked in 10% bleach laid out across the interior base to reduce the chances of contamination. That's a little bit of overkill, but I'm also using a home made laminar flow hood with less than perfect design.

Best,

Rob

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Re: Sowing Seed Question

Post by RandyT » Wed Aug 31, 2011 5:17 pm

Z-
My neighbor's setup:
Laminated plywood sides, top, back and bottom. Plexiglass slanted front that lifted out to place materials inside. The edge of the box that seated the front was "padded" with foam strips to make a more airtight seal. The weight of the plexiglass was all that kept it in place. Below the slanted front, was the glove setup, I think it was plexiglass with gloves set into holes and I think siliconed in. The slanted front makes it much easier to see what you're doing in the box.

To use, he'd remove the slanted front, wipe the entire inside of the box with clorox, 100% (I think 10% will do fine), wipe down the empty flasks with clorox, place them into the box, put in sterilizing implements after wiping (sterilizing implement can be alcohol/flame 3x or soaking in bleach solution for 10 min. and are for sterilizing tool once inside the box) tools (spoons, tweezers, scalpels, razor blades, whatever.) Replace the front. You'll need to put in everything you'll need to flask before you close the box, or repeat the sterilization process every time you re-open the box. Let the box sit for 10 min. or so to let any dust particles settle before starting any flasking.

If you're doing this just for yourself as a hobby set up, you may consider using glove set into a traslucent covered container like a Rubber Maid storeage box. This can be done with a minimal amount of time and money. If you enjoy it, you can build a glove box or purchase a sterile hood.

I wouldn't bother with a uv light in a glovebox. It isn't as effective as a bleach wipe and cannot be used with you around the box. UV lights will give you a very bad burn.

Have fun!!!

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zxyqu
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Re: Sowing Seed Question

Post by zxyqu » Thu Sep 01, 2011 6:36 am

Thanks Rob and Randy, that's all quite helpful. I was originally planning on using the lab I work it to do all the flasking (I even got the big whig's blessing), but then he took a new job, so the lab won't be available anymore.

I mentioned the UV setup, as I've got a couple in lab that come home with me, and if you cover them up, you dont have to worry about a burn (though do I have a story of a guy in our lab just hanging their head over the gel lamp... talk about raccoon eyes!). And the setup you're describing is pretty much what I had in mind, but had some great tips. Anyway, I'll let you know once pod meets bleach, and thanks again for the help
>> J <<
You know it's a great forum when you search for "bellina" and it returns this from the server:
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hhartung
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Re: Sowing Seed Question

Post by hhartung » Thu Sep 01, 2011 10:35 am

zxyqu,

you don't need UV in the hood. It's dangerous. And never ever work with the UV switched on, covered or not covered.

Helmut

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zxyqu
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Re: Sowing Seed Question

Post by zxyqu » Thu Sep 01, 2011 10:43 am

I know, I was commenting that I could sterlize the hood, covered, then turn the UV off, and use it. Given what was said above about the bleach, and letting stuff settle, I'm somewhat partial to that.
>> J <<
You know it's a great forum when you search for "bellina" and it returns this from the server:
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orchidgfs
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Re: Sowing Seed Question

Post by orchidgfs » Thu Sep 01, 2011 2:48 pm

zxyqu,Ive been using a glove box that I built in 1968,its 24by.20ins square,made out of 8th of a in pressed board ,replaced the bottom because the bleach eates it up after a while,glass on top with the box cut at an angle,and a door on one side about 8in sq,and a lamp on the top,this is a little large for me but I think Ill still use it,have fun,glenn,ps,I think 50 bucks or so should be enough to build it

stock
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Re: Sowing Seed Question

Post by stock » Sat Sep 03, 2011 7:44 pm

I directed a tissue culture facility and chromosome lab with many hoods containing UV lamps. They were never turned on for use in the many years I directed the place for the simple reason that they are not effective. Their effectiveness goes down rapidly after a little use and even when new they kill very little in the way of mold spores on the work surface of the biohood. Forget them and just wipe down the surface of the hood with a good disinfectant like physan solution.
Dean
Albert Dean Stock, Ph.D.

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