sorry for the spelling

 

I used to DIY with a 10g on its side, some cardboard, some rubber gloves taped to the cardboard, and a can of lysol. That was for growing mushroom mycellum so I dont know if it would be enough for tissue culture. Same principal tho.

 

from make...
http://makeprojects.com/Project/Home-Mycology-Lab/242/1

also from a fellow aquascaper
http://www.plantedtank.net/forums/plants/149185-jeffs-foray-into-ptc-first-failure.html

 

thanks very cool------ funny---- I want to try to build a small hood if it looks like a duck, and walks like a duck , it will be a duck..

 

thanks for the link clark seams to be a lot of srum fans here lol

 

Just an FYI on hoods, we use several diff types in Pharma, depending on the purpose. Traditional fume hoods (ie chemical hoods) are a designed to prevent exposure if the operator to fumes by having a sash a the front for air intake, and a number of vents at the back (at bottom, middle and top) to pull off the nasties.

Microbiology labs use Biological Safety Cabinets; same general front and back to them, but an added channel of out-vents right inside the front lip of the hood, so that anything floating in through the operator's sash gets sucked out immediately and can't contaminate the plates.

The third type I've seen are Laminar flow hoods. These are used in Sterile Manufacturing, and work in reverse from most fume hoods - thy have a hepa-filtered fan on the top of the hood blowing air in, and the air blows out over the operator. This works to prevent contaminants from approaching the clean surface.

To maintain aseptic conditions, you'd probably be wanting the third type, rather than the first. How you'd DIY that I don't know; I get to buy all mine from suppliers...

 

thanks shorts--buying one i would consider but i have a feeling it make dent the the wallet
and be to big for expermenting tissue culture of aquatic plants
But your info is greatly appreciated

 

Yeah, I doubt anyone would be picking up anything I play with professionally for aquatic hobby purposes. Mostly I was hoping to give you a bit more I sigh into the diff types so you could search for designs more effectively.

GL with the project. I have heard of of amateur microbiologists setting up hoods at home on the cheap, so it can be done. And beyond anything else, cleanliness is of the utmost priority.

 

It might be worth seeing what sort of surplus the local hospitals/universities/governments around you have.

I think that you can find a lot of older/unused lab equipment from the UC's around here for sale to the public a couple times a month. I imagine it would vary depending on the institution, but I don't think it would be uncommon for the larger ones to have some sort of surplus system.

We had a tissue culture hood in my lab, but never used it, and had to send it to surplus to make room for some other equipment. I believe it had some UV lights for sterilization, but I'm pretty inexperienced with that, and we never even got it up and running.

would there be much advantage to using tissue culture for aquarium plants? I was under the impression that most of them propagated fairly easily.

 

You basically take a box and a HEPA filter and have the outlet blow through the box gently. I believe that it's not necessary to make an actual bio cabinet but it can be done. Oh and I'll get back to you VERY soon on the PTC stuff. Spring break is here and I've just prepared a fresh batch of medium.

And to answer your question lochaber, PTC is great for slow growing, valuable plants like crypts, erios, and anubias. My personal goal for PTC is to culture some rarer crypts and hopefully work on some bucephalandras in the near future. I have a thread here on it but I have had to put up my project because school's been a time hog.

 

I wasnt going to use the buce word but as u know there are some painfully growing plants
If you have a rare cyrpt for instance you might get one offshoot in 3-4 months-inculture you can trash that plant and clon a dozen or more in a shorter time span --time is money!!

 

Jeff/Buce thanks, that makes sense now that I think about it.

I think I had most heard about cloning plants in regards to venus flytraps, which I thought was to prevent poaching wild species (which are endangered), but now that you mention it, and I think about it, I guess there are quite a few aquarium plants that breed a bit slower then duckweed...

:/

 

And thats the best word yet propagating species is all about speed and not taking plants from the wild !!

 

Have you started your project yet? If you don't mind me asking?

 

Gettin allmy ducks in a row --Ive took on to many projects at the same time Jeff but I will be in touch when im ready to go i think its important that a bunch of us crash and burn together LOL NOT

 

Bruce, I am curious about tissue culturing plants like your talking about. Is there a good reference site that I could check out to learn about culturing?

Specifically one that's safe for work, lol.

I'm not looking for the theorum behind it. I'm looking for the how-to, nuts and bolts of it all, with a little biological theory, science-stuff thrown in. I haven't taken a science class since the 90's. My wife however, can fill me in on the important details (pre-med degree, biology and chemistry and whatnot)

Thanks for the info!

 

For tissue cloning 101 you can google up on u tube some great and some bad movies to watch thats where i got my feet wet Im hoping we can form some sort of support group to helpall thats interested along with what i think will be a rewarding project!!
Stay tuned
Bruce

 

Well, just to get back to you. Lucky me it turns out I have a large HEPA filter ready to go at my home. It's a "medical clean room" quality product apparently with .3micron filtration at 99.97%. Though rather than use it in a cabinet type set up I think having it running for several hours in a small room will be pretty sufficient. My largest hurdle right now is probably creating sterile tissue since soil is nasty nasty stuff in terms of microbiology...

 

I use a broken 55g turned on its side with a fan blowing on top of the tank to prevent anything from 'falling' into it. Now if I can just get the chemicals to actually tissue culture...

 

You don't want any air movement at all if you're going to go with a clean box. Unless the air has become functionally sterilized by going through either a scrubber or HEPA filter it's filled with pathogens and contaminants. Just make your movements very slow and deliberate, keep your mouth shut/wear a mask, and turn off any fans/close doors/seal drafts in your room.

http://www.youtube.com/watch?v=zd0iVJrQwyY this is good watching.

 

jeff cant you integrat that hepa in some sort of box with plexy in front you do need that air movement in front but not where you work

 

I could, I have all the materials I need but since I'm limited on actual culture materials at the moment I don't want to end up contaminating all my cultures using the filter I haven't really tested. I'm just going to have it operate in my room for a day prior to my culturing which should reduce contaminants to a good degree.

 

Here is the one most successfully and coommonly related one to the subject you are looking into...

http://kryptokoryne.aquaticscape.com/

 

That was a good vid im almost ready to diy a flow hood

 

Tissue Culture 101 Part 1

Part 1
Here's the deal, I have done tissue culture in college many years ago.
Here's the process
1)You're taking a growing tip of a plant, sterilizing it, putting it in a medium (a chemical mix) that initiates undifferentiated growth (much like cancer).
wait x number of weeks
2)Split up that growth into small pieces, put those pieces in a different media that encourages differentiated growth (plants with shoots and roots) all still sterile
wait x number of weeks
3) grow those pieces on in a still sterile media
wait x number of weeks
4) Once those pieces are little plants carefully move then into a somewhat not sterile place (greenhouse or grow room)
wait x number of weeks
5) Overtime as the plants get larger and more resistant/harden off they can be exposed normal conditions, at that point planted or sold or whatever.

 

Tissue culture 101 part 2

Sterile procedure is the key
You want little air movement or as was stated before movement of air that is absolutely sterile as coming from a HEPA filter
You can get away with just a room with little air movement, or a tank on its side wiped down. but at each step of the way you increase the chance of contamination.
Even preparing the pieces if not done right can cause contamination.
Or at any point during any point of the transfers I described in part 1
It takes careful slow movements not to kick up any particles and as clean a room as possible.
The bottom line is maybe 6-7/10 times (just making the numbers up) you could do it in your kitchen if you were careful but the first 3-4 times might be a failure, thats why labs make sure everything is prefect so they get it right every time.

Its a pretty easy procedure, with very clear simple steps its just getting the technique down
Science isn't all science its an art too

Part 3: How I would set up a TC lab or should i just skip to part 4
Which I'll probably anyway do if nobody cares

Part 4 why do tissue culture at all when you can grow plants in a tank or emersed so well?

 

green man Some crypts and other rare plants spread by rizom or runners painfully slow--
growth is growth so why not clon 10 20 30 plants from one rizome instead of waiting for one plant to divide? Your expertise would greatly be appreciated why not watch this post and help us tenderfoots when where ready?