Flasking 101 - A how-to guide

[Zozzl]

If you have always wanted to know how to flask your own orchid seeds but did not know how to get started this thread is intended as a guide for orchid enthusiasts with no prior experience in flasking. We hope to get you get going so you can have the thrill of watching your seeds germinate and grow

Many of the people who post on this forum have a lot of expertise and have perfected their techniques so they can flask easily and quickly. We are asking for those folks to participate in this thread to help others get started by posting a brief note about their flasking techniques and after we have a nice collection of those posts we can go back and break it down into steps so that it is very easily understood.

Hopefully, we can break flasking down and demystify the steps so anyone who reads this thread will be have no problem getting started. There is already a lot of info on the propagation forum but having it one place will enable OB'ers to print it out and use it as a guide.

So.....anyone with flasking experience, we need you!! Please consider adding your valuable post to this thread. If does not have to be long but just go through briefly what you do to get those seeds into those flasks and growing We will be breaking those posts down into steps to make it crystal clear but we cannot do it without YOU.

Is there a brave soul who would like to go first? :

[Gin]

Here is what I did ,not scientific

Things You will need .
Pressure cooker ,
Flasking medium I ordered mine from Gallup & Stribling (Hills ) all you add is boiled distilled water.
Gal. Distilled Water
Bleach
Foil
pint jars with lids and bands or better yet regular flasks .
spray bottle
10 gal.
Aquarium
Latex gloves not the thick ones ,
Plastic with holes cut in for your hands , Duct tape to tape the gloves to the holes and plastic to the open part of the aquarium .
New single edge razor blades , or Exacto knife
soft tooth brush
2 small plastic containers that will fit in the aquarium .
----------------------------------------------------------------------------------------------------------------------------------------------------------------
Add Hills to boiled distilled water mix well , put a couple of inches of water in the bottom, of the pressure cooker , place jars in it, fill 2 inches or so with the mixed hills . Have foil squares cut place them over the jar mouth put lid over the foil then put band on not tight you do not want the jars to seal when cooling .Seal pressure cooker, bring up to 15 lbs. for 15 minutes after jars are cool tighten bands . Set aside for 2 or 3 weeks to be sure there is no contamination before going to step 2 ..
Step 2 if no contamination shows up . Set up glove box (aquarium ) tape plastic to the open part with the gloves attatched .
Spray the $%^%$ out of the inside with 30% bleach water I also spray the outside of it. Place containers in the box respray .Also spray the outside of the jars and under foil edge before placing in glove box once in respray with 30% bleach water .
Green pod is the easiest , watch for signs of yellowing . remove it . Use the tooth brush to scrub the outside of the capsule with 5% bleach water for 5 to 10 minutes can be done out of the box rinse with boiled distilled water . wear gloves sprayed with 5% bleach water . place in the glove box respray gloves , slice pod length wise . Open jars (risky part ) tap out seeds into the jars place foil over jar opening then the lid screw band down tight . Then wait
I probably left something out , it is just a lot of tinkering and making sure contamination is not brought into the box . gin

[lambelkip]

thanks Gin, I was about to look for a flasking tutorial. I have some seed pods that are just about ripe.

[Zozzl]

Great job Gin!

We need as many posts and techniques as possible so step right up folks

[Gin]

I have not done dry pod (already opened ) there is a good thread on here about it .
Better ways then how I do it, would be a lot easier with the proper equip . Good luck on your pods Come on guys and gals we know you are out there need all the varied info.available Gin

[10010100102]

Well, I use your basic glove box technique for the actual flasking process, but where my method really differs is in the medium preparation. I don't have a pressure cooker, and I'm not about to go buy one just for this. So, here's what I do.

I make up my medium in one pot, and while I'm doing this I bring a big pot of water to the boil right next to it (with all my clean, empty flasks and lids on the counter nearby). I also get out a cooling rack and set it down on the counter next to my pot of boiling water.

Once the water is at a rolling boil and the medium is ready, I start working. I put one lid into the boiling water, and use a pair of forceps to pick up a flask and suspend it in the water. I wait a few seconds for the glass to sterilise, and then, working quickly, I pull it out of the water and ladle (or spoon) some medium into the flask. Then, I bring the flask back over to the pot of boiling water and use the forceps to submerge as much of the flask as I can without getting any water actually in the flask. I then pull the flask out of the water and set it on the cooling rack (I think having the inside of the flask so hot from the boiling water creates a draft of warm air up and out of the flask which prevents contamination during this step if you work really fast). The trick now is to fish the lid out of the pot and screw it onto the flask as quickly as possible without scalding yourself! Line all the flasks up on the cooling rack as you do them; this way they can cool more evenly and when you're done you can move them all in one go.

The disadvantage to this method is that when the air inside the flasks cools, it contracts, building negative pressure on the inside of the flask. This makes it so when you open it to put your seeds in you have to do it in a sterile environment (I've heard of flasking techniques that can be done out in the open by creating a draft out of the flask while you work), otherwise when you open the flask it'll suck in all kinds of nasty stuff.

I think this is a great idea for a thread, I'll definately be checking back on this one to see what other people are doing. I'm especially curious to hear about some home-made medium recipies, especially if the results are comparable to those with commercial medium.

[King_of_orchid_growing:)]

I'm just starting out with flasking, so I can't contribute too much.

I can add that you can sterilize your plant material, tools, and work area using a variety of disinfectants.

1. 30% bleach water is one way
2. 70% isopropyl alcohol is another
3. 3% hydrogen peroxide
4. Finally there's calcium hydrochloride or sodium hydrochloride (not sure which one) - note: (some species of orchids are sensitive to this, do your research)

Note: It is a good idea to disinfect anything you can think of. Having to replate after a contamination is time consuming.

You can sterilize your flasks using any of the above mentioned ways in addition to:

1. autoclave (expensive)
2. pressure cooker
3. steam from a pot
4. oven (350 degrees F to 450 degrees F)

Different sowing media are also available. If you contact a lab they can help you with the type needed. They will also discern the difference between sowing media and replating media.

Depending on the species you are growing, you may want to consider the hardness or softness of your media. If the plants like moisture, soft media with as much condensation as possible is good. If the plants are dryer growers, a more solid media is better.

You can also make your own media using stuff such as:

1. potatoes
2. carrots
3. oat meal
4. coconut milk
5. pineapple
6. bananas
7. beets
8. radishes
9. corn meal
10. agar

Depending on your species, you might want to know if the seeds should be sown in dark or light.

[King_of_orchid_growing:)]

I forgot to mention, there are two methods to in-vitro (flasking) sowing.

There's asymbiotic, which many people are familiar with.

Then there's symbiotic, which very few people (including me) truly understand. This is not recommended for somebody starting out.

Symbiotic in-vitro seed sowing requires you to understand the symbiotic relationship that orchids have with a certain kind of fungus generally called mycorrhizal fungus.

These mycorrhizal fungi infect the living root tissues of the orchid, and after a while, the orchid roots starts digesting the fungus.

In order to do the symbiotic germination method, it requires you to isolate the fungus and grow the seeds with the isolated fungus species. There can be trial and error involved with this as the relationship between orchid and fungus can be species specific.

[Royal]

I sow seed in my kitchen with a homemade glovebox and very few bought items. I've only been at it for about two years, but I had moderate success. Just to clear a few things up, if we're really talking about "Flasking 101" then we need to start with a broad overview for the total novice. Also, we should probably stick to basic technique we have learned by doing, not what we've heard or read. Lastly, let's assume that "Flasking" in this context means the germination of orchid seeds asymbiotically and growing in assumed axenic culture.

So here it is from the top:

Orchid seeds are tiny. They don't germinate and grow like "regular" plants - we all know how slow some can grow. In nature, a seed needs to find the perfect place that meets all its very specialized requirements. A mixture of nutrients and/or other chemicals can be used to coax the seeds to germinate and grow. This nutrient media can be used as is, or is often solidified with agar. The catch is that this media grows lots of things well - mold, fungus, bacteria, algae, and oh yeah... orchids. Guess which grows the slowest.

To keep unwanted cultures from growing on your media, it must be sterilized and must be well sealed inside a container (flask, jar, petri dish, etc.) to maintain sterile conditions over a long period of time (did I mention that seeds grow slowly?). Sterilized is not the same as Sanitized! Boiling temperatures do not kill heat resistant spores and cysts. The media, the flasks, lids, and tools should all be autoclaved or pressure cooked @ 15 psi for 15 minutes.

Seed can contain contaminants as well so they too must be disinfected before putting them into your nice sterile flasks. This is the tricky part because since they are live tissue, they cannot be heat sterilized. We have to just live with sanitizing here. Finding the right balance between killing all contaminants and not harming the seed is the key. Many seed "steriliants" can be used successfully, but household bleach mixed to 1:10 is a good place to start. It's cheap and available everywhere.

Transfers must be performed in a "sterile" environment. (I keep putting sterile in quotes because the only things that are truly sterile are the items put through the p-cooker/autoclave) Big labs and breeders use a hood that forces high efficiency purified air (HEPA) through a workspace to eliminate contamination from entering. All work is done in this stream of air flow. Gloveboxes work by eliminating any exchange of air at all. Once sealed and cleaned they should keep out contaminants. Others have successfully sown seed under a towel saturated in bleach, over a steam bath, or even between to panes of glass. I recommend making a simple glove box.

Finally, the issue of growing on and culture. This is an area where I need the most improvement. I figured I'd worry about it when I actually got seeds going, but I'm learning that culture is integral to success here. I'm always learning, but I've learned the most by doing. I'd like to encourage you all to just go ahead and jump in. My set-up is anything from fancy, and totally make-shift. If I can do this, so can you.

[Edited to add...]: One more thing for the newbies, you'll hear terms like "replate" or "mother" flask. This refers to the fact that orchid germination medium is specifically made for germination. Once the seeds turn green and start to turn into little plants in the mother flask, they might need 1) more room to grow, 2) fresh new media packed with nutrients, 3) different flasks equipped with microscopic vents to allow for gas exchange, or 4) a different formula all together with or without certain ingredients or growth regulators. We transfer them at this point to space them out more efficiently and to give them optimal food for the 1 year + that they will grow in vitro. The need to replate is scary for the newbie sometimes and he/she might want to try to get by without it. Don't sweat it. Safety in numbers - just make more of everything than you think you'll want or need. When you get 50% contamination, you'll be glad you made doubles of everything.

[Zozzl]

Great posts, Joseph, King and Royal!

Thank you