Advanced_Orchid

ADVANCED ORCHID FLASKING

After using the "Simple Flasking" for awhile, I had reached the problem of "replating" ( seperating the protocorms into other flasks to enable them to grow on) and while the process for seeding was found to be very good, you will remember it entailed carrying out the process under a Tea Towel and without being able to see what you are doing, would make it impossible to place the protocorms in the flask.

So the next step was to make up a "FLASKING CHAMBER". I have seen a number of different ideas as to what different people have made up. I chose to buy a 3ft Glass Aquarium Tank which I had two round holes cut in the side; to this I glued a very long glove to the hole so I could place my hands through. I turned the tank upside down so the opening was downward, placing this on a sheet of white malamite.

Sterilising Here lies the success or failure in flasking! The inside of the chamber must be sterile at all times and this is obtained by spraying it well before you commence; I started using bleach ( 20% bleach /80% water) but found after a number of months the fumes were making me feel sick, so I changed to using Methylated Spirits (ethanol) (70% ethanol /30% water) and this also drys quick where the bleach left the chamber wet all the time. For the flasks I found after using a number of chemicals I came back to dipping them into a bleach Solution of ( 10% bleach /90% water ) these are placed wet inside the chamber. Tools - forceps, etc are also dipped into the bleach solution before placing in the chamber; many people place them into a container of the solution inside the chamber to sterilise in-between flasks, I had a small spirit lamp which I flamed them by dipping into neat Alcohol. I found this a better way to cut out any cross infection from flask to flask. This action made the inside of the chamber Sterile and I had very little fungal problems.

Green Pod Flasking you need a container which contains neat Methylated Spirits or Bleach, a tooth brush to dip and scrub, a scalpel to cut the pod, a small square of glass to cut on( remember all of this is done inside the chamber; and also everything that goes in must be dipped in the solution first) I have seen mention of the need to scrub for 30 mins, I have always found about 5 mins is ample ( while the inside of the pod is sterile, the outside is covered in dirt and fungus, this must be removed), you then cut the pod in half with the scalpel; unscrew the lid of the flask and with the blade scrape a small amount of the seed over the top of the medium ( try not to over do it) if you have dropped it in piles then gently with the blade ( or a piece of stainless steel stiff wire that has a bend on the end) move the seed over as much of the medium as you can. Place the lid back in the Flask; job done.

Dry Seed Flasking I still used the same process as I mention in the "Simple Orchid Flasking"; the only change is I don't have to use the tea towels.

Replating Protocorms observing all the protocol for sterilising as explained in "Green Pod Flasking", we will assume you have the flasks in the chamber ( both the "Mother Flasks" and the flasks you will replate into).

The idea is to allow the plants to grow quick and strongly and if left in the "Mother Flask" ( a flask that you sow the seed into is called a "Mother Flask") they will not make much progress, so you do one of two things, 1. Spread; by taking an amount of the protocorms and just spread them on top of the medium. 2. Place a set number into a flask by actually planting them into the medium. I prefer the 2^nd way as I like to place about 15 to 20 protocorms ( the number depends on the size of the flask you use) in a flask, I just push the end of the protocorm a little way into the medium to hold it upwards ( the top of the protocorm usually has a point) this allows the protocorms to grow into plants and progress. Im often asked at what size do you replate; I recommend that you start when the protocorms are large enough to pickup, about the size of a match head. Of course you can replate at any size you want too.

Flasks & Mediums

Flasks -I use two sorts of flasks, an Unvented one for Mother Flasks and a Vented one for all Replating , a vent is necessary to allow the gas and air to transfer from the flask. There are a number of ways to vent a flask I use 250ml glass jars with Plastic lids ( auto-clavable) I drill a 1/4" hole in the lid to the side; you can buy small round Teflon patches or place some plastic tubing in the hole and fill it with non-absorbent cotton wool or just stuff cotton wool in the hole. I have used all three mentioned, but have now come back to the tube with the cotton wool plug, I'm finding it is 100% fool proof.

Mediums - For seeding I have found Sigma's "Phytamax" Maintance Medium (P6688) used at either 1/2 or full strength the best, others are Vacin & Went and Murashige & Skoog either full, 1/2 or 1/4 strength, use these for "seeding".For replating Sigma's "Phytamax " Medium (P1056) has been found to grow most genera very well, and other companies also make up proprietary mediums to much the same formulas all over the world. I also make up a few mediums using general fertiliser which work quite well also.

Sterilising Medium - Most proprietary mediums you purchase are in powder form and have everything included, apart from Agar ( I use 7gms/litre, this gives a firm medium) you just add the powder to distilled water and follow the directions in "Simple Orchid Flasking"

Laminar Flow Cabinet

Because I started to do flasking for my local Species Society ( to provide "plant of the Month") I have moved up from the chamber which restricted the number of flasks I could have in it at one time, so I bought a "Laminar Flow Cabinet", this enables me to work without restriction regarding flasks and also with the front being open I can move around better. I have a Bunsen Burner inside and now flame all the tools; any fungal problems I did have ( and believe me if you think you can run a lab 100% clean, you are kidding yourself) with the use of the B/burner and 70% Ethanol spraying of the cabinet, I have most of the problems that I have experienced to a minimum.

Oct. 99 up date - I have now purchased a "Glass Bead Steriliser" from India and this takes the place of the "Bunsen Burner"; while the b/burner did a great job, there was always the chance of either a fire or the flame going out, so for safety sake I felt it was important to change over. While using 3 sets of tweezers I am able to have one set in the steriliser, one set cooling off while I use the 3rd set. Since using the steriliser I now note that fungal is almost a thing of the past. Click onto Lamina Flow Cabinet above to see how it is setup.

Dec 2000 update - Dry Seed - I now have found an excellent result for sterilizing Dry Seed. I make a solution of 100ml of Distilled Water and I add 5ml of 'White King' ( contains 42g/l of Sodium Hypochlorite) or (Clorox) to a total of 105ml, I don't add any wetting agent as 'White King' already contains it. Using very small bottles (5ml) with screw top lids, number them for your records. Add the dry seed first, when all the bottles have been filled with the seed, I then add the liquid (screw the lids tight) so I can then have them all about the same time limit (5min) remember to shake the bottles a number of times and then place all the bottles in the bleach solution before placing in the cabinet. I let the liquid/seed settle and using a pipet ( a tapped glass tube which attach either a rubber squeeze ball or a retracting pump) and draw most of the solution out, adding Sterile Water ( distilled water auto-claved for 20 mins.) too wash the seed: this is then tipped into the flask and left for up to 20 mins. To settle on top of the medium. I tip the flask to the side and using the pump I take the excess water out. Re the Pipets- I have a numbered rack ( numbered 1 to 20) and place the pipets in each slot, this way I use one pipet per flask, you don't end up with a mixure of seed germinating in a flask. Remember the glass Pipets must be auto-claved to make them sterile; I place them in a oven bag, sealed with tape and usally place them on top of the flasks in the Pressure Cooker. The bag must also be washed as all other tools and flasks are before placed in the cabinet.

Want lots more information on flasking?????

I would suggest you obtain a very up to date book titled "Asymbiotic Technique of Orchid Seed Germination" by Aaron Hicks from The Orchid Seedbank Project in the USA. This book covers from A to Z on flasking and covers lots of old and new ideas , mediums, sterilising etc. etc. Available from the Author Aaron Hicks direct ( go to our 'Links' page and click onto The Orchidseed Bank for details and prices)

Flasking Cool Growing Orchid Seed by Bob Hamilton. A very good article on flasking, I feel is applicable to most types of Orchids.

Lotte&Thomas's Flasking Site - Another excellent site describing lots of Orchid Propagating Techniques, including 'Flasking under Steam'.

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