Fundamentals Of Flasking

The notes below are a guide to how we flask and this works for us. If you have any tips please feel free to contact us I would appreciate any information.

Flasking is not for the faint hearted,

first you need

seed.

flowering plants to pollinate

once pollinated seed can take from 3-18 months depending on genera

then plants will stay in the flask for normally over 12 months before ready to pot out

CAUTION

ALL INFORMATION IS IN GOOD FAITH IT IS YOUR RESPONSIBILITY TO READ CHEMICAL AND EQUIPMENT LABELS ETC..... AND FOLLOW ALL SAFETY PRECAUTIONS GIVEN ON LABELS. ALWAYS FOLLOW THE MSDS GIVEN ON CHEMICALS AND EQUIPMENT

IF YOU DO NOT FEEL CONFIDENT OR DO NOT UNDERSTAND ANY INFORMATION ON LABLES OR MSDS DO NOT PROCEED

Some History

Orchid seed do not have a nutrition pack to survive and grow like other plants, orchids need a fungus to grow, this fungus is found upon the root of orchids. To propagate you would disperse seed from a purple cattleya around a white cattleya and visa-versa and when/if the seed grow and after time flowered it was obvious to what plant was what. Today the mix/medium has chemicals that replicate the fungus needed by the the seed to grow.

Flasking EVERYTHING MUST BE 100% STERILE. The use of a laminar flow cabinet is preferred. You will need a dedicated room with sink, vinyl or tiles on floor (to help keep clean and help wipe spilt chemicals), power, racks to hold flasks, washable chair and air conditioner running 24/7 and grow lights for 14 hours every day. if this is not possible then rethink about doing flasking

Picture of part of our Lab holding around 2000-3000 flasks

You will need a few things to get started

Autoclave
Flow Cabinet
Scales and balance
PH tester (not paper strips as can vary with charcoal in the mix)
Microscope / stereo microscope
Bottles with and without breathers
Blender (not to be used for food after)
Alcohol @ 70% (not metho as long term use may cause eye damage)
Tools for replating and cutting seeds (scalpels, long tweezers, micro spoons, loops, cutting plate etc...)
Lab coat or clean old clothes as bleach and wife (sometimes) is not forgiving
Spray bottle for alcohol
Paper towel
Small bucket for dunking flasks into bleach
Labels
Soap
Sterile wash
Clock/timer
2 x ice cream containers
Gloves

Laminar Flow Cabinets

You will need medium/mix for your orchids to grow on there is lots and lots of different recipes and these are also genera orientated. in Australia we are lucky to have a few different suppliers of pre-packed mixes. Westerns Lab, bio tech, sigma, and others advertise in the orchid review books every month.

HERE GO'S!!!!!

Make up a batch of mix, I use boiling water as this helps to set the agar you will need around 25mm deep mix in each bottle this can be a bit less for seed sow. Autoclave your flasks and allow a minimum of 20-25min @121c or just above for approx 12-24 flasks per batch, this is something you will need to play with, too much will burn the mix and too less will not sterilize and will contaminate the mix

When removing flasks use two clean cloths or paper towel one to hold bottom and one to tighten loose lids as you pull them out of the autoclave, also be gentle and do not shake. You do not want any of the mix to touch the filter otherwise you will get contamination. Allow to cool and write date and type of mix etc..... on lids. move the flasks to your lab these now need to be left under lights ready for use in 1-2 weeks. This is to allow time for any contamination to grow if something went wrong

Tip:- mould that is visible within 2 days and is growing on top of mix, it is a good chance that the breathers are not sealing/working. If mould is taking longer 1-2 weeks to develop and mould looks like it is leeching out of the mix its a good chance the flasks have not been autoclaved properly

Autoclave your clean tools i do this for 25min this time depends on what mass/size volume is to be autoclaved. You will need to turn on you flow cabinet and spray and wipe out inside and around front with alcohol when complete turn on UV lights and remember you can get sunburnt very easy from UV lights. it is best practise to run this for approx 20mm before use this allows time for purging and UV to tack affect.

Get your seed sterilizing under way

Wash green seed with a detergent in an ice cream container with a new tooth brush for min of 2 min
Now wash seed in a ice cream container with a 50:50 mix with "Aldi" bleach and water with a new tooth brush for 2 min
Make up a 50:50 mix of bleach and water in a bottle and fill to the top
Put washed seed in bottle for min 10 min

Orchid seed under Microscope

Make up bleach solution in bucket 1:10 bleach to water this is for dunking your flasks into before putting into flow cabinet. PUT A TEMPORY LID ON BUCKET

Tip:- Bleach has a shelf life if not used ditch after 6 months and always keep the lid on, All of your movement around work area should be slow and gentle as to reduce air movement

Turn off UV lights when time expires, once your tools are ready take them out of the autoclave and place them in the flow cabinet

Scrub your hands and arms with soap and then sterile wash then spray with alcohol, put gloves on and spray again with alcohol. this makes a sterile barrier for you to work. Remember to not touch anything other than if it is in the flow cabinet or if sterile as IT WILL BE CONTAMINATED, If you drop your tools in the cabinet or if they touch something other than the direct work area discard and use another as they may be contaminated

Dunk the flasks, seed bottle and alcohol sprayer into the bleach bucket one at a time making sure that all the outside surface are in contact with the solution and place them into the flow cabinet. Spray the work area/surface of the cabinet, remove the seed lid and a mix lid, cut seed capsule and place some of the seed onto the mix in the flask remember that tools, inside, rim and lid of flask and seed must be kept from being contaminated at all times. i make up two flasks of seed with different tools as to reduce the chance of contamination to one flask

this seed is not ready

this seed is no good as you can see with the seed they were picked too early

this seed is ready

Place lids on flasks and tighten when finished remove all flasks and label with date, type of seed, batch number etc......

Clean all tools, work areas, turn off cabinet and drain water and clean autoclave

Put flasks under grow lights and cross your fingers. If all goes well sometimes whitish seed turns green in days but can take up to 12 months to do anything. This depends on the type of orchid

Pictures above are the different stages of flask development

some times you will find that not all/if any seed are viable and will not germinate and this has a lot to do with plant parentage and time seed capsule was picked (too early and seed is not viable)

Once you have growth to replate do the same as above but always remember to change or keep tool sterile as you go and do not over crowd your work area as this affects air flow from the HEPA filter

Happy Growing