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4 Materials Needed for Plant Tissue Culture

Plant Tissue Culture Materials Can be From Repurposed Plant tissue culture has many aspects of the advanced scientific world.  Autoclaves and chemicals being the most pronounced.  There’s no reason you cannot practice plant tissue culture at the hobby level and not have high returns in plants.  Achieving success may sometimes mean substituting or rethinking what you are doing, but it can definitely be done.

These materials and supplies will get you through plant tissue culture from start to finish.  No question about it.  These same materials and tools have created tens of thousands of plants for me quickly.  They can do the same for you as well.

Workspace

Aquarium or Plastic Bin or Tote
Glovebox
Flow Through Hood

These make for a sterile area to work with your cultures.  All work must be undertaken in a place that is free from falling dust and other contamination.  The best tool is a laminar flow hood but may be cost prohibitive to the home hobbyist.  I have plant tissue cultured tens of thousands of plants, or more, with a 20-gallon glass aquarium laying on its side.

A glovebox can also provide a suitable workspace.  They can be built rather easily but are not necessary.  If you cannot get clean cultures with a rubber tote or aquarium you should try a glovebox, however, great success can be had without a Laminar or glovebox.

 

Autoclave for Sterilizing Vessels

Autoclave or Pressure Cooker
Microwave

Vessels must be sterile.  An autoclave makes steam heat in excess of 220 degrees which kills nearly all contaminants.  Autoclaves are reliable but expensive. A great alternative is a 23-quart pressure cooker. I’ve used a Presto model for an autoclave exclusivenly with great results. Any size will work but larger sizes will probably be your best bet as you can make a whole batch of media in jars at one time.

As a sterilizing alternative for small batches, a microwave can be used.  This is usually best for explant initiations with small amounts of media inside.  Heat briefly, let it fall, and boil it briefly.  This is done several times to sterilize the media.  It is tedious but it works.

 

Vessels

Baby Food Jars
Small spot bandages for covering the hole in the original lid.
Magenta B caps for reusable lids.
Commercially available test tubes and vessels.

Vessels hold your media and prevent contamination of your cultures.  They need only be of a suitable size and easy to work with, as long as they have some sort of lid.  Even at that, aluminum foil and a rubber band can top any glass vessels suitable in size and withstanding the temperature of the autoclave or pressure cooker.

Baby food jars with the original metal lids as well as Magenta B caps can be used.  Magenta B caps are far easier but the metal lids can be used by poking a small hole in the lid and covering the hole with a small shaving spot bandage.  They are usually available in round or square shape at local pharmacies.  Sponge cubes can also be rolled and pushed into larger holes but prevent vessels from being stacked.

There are many commercially available containers for plant tissue culture.  Most small vessels are best for the initiation phase and larger vessels work well for large culture volume.  Some tall plants like certain bamboo species can benefit from tall vessels.

 

Staying Sterile and Safe

Antibacterial soap is good for cleaning explants in the first step of sterilization.  Not a bad idea to wash your hands well before starting culture work too.  Any will work.  Some are better than others but overall any good antibacterial soap will work.

Nitrile gloves or latex are important.  You want to keep your explants sterile while protecting your skin.  Wear some form of nonporous medical glove while handling chemicals and plant material.

Wear eye protection.  Something that protects your eyes and is comfortable is recommended.

Masks that keep your breath from pushing air into the workspace can be a huge improvement in getting clean initiations and replates.  It doesn’t matter if it’s the work type masks or the medical type facemask.  Keep your breathing restricted and you will have fewer contaminants.

To keep dust from clothes an oversuit would help.  Tyvek suits are available and can help reduce dust and contaminants from clothing.

 

Measuring Scale

Accurate measurements are important.  Lots of materials are weighed to make solutions and media.  Accurate scales down to milligrams for weighing chemicals are reasonably affordable. Find one that has calibration weights with it.  Calibrate it before each use. 

 

Measuring Spoons and Cups

Smidgeon” measuring set for quick measurements.
Graduated container 1 liter or larger to measure liquids.
Paper condiment cups for weighing powders.
Plastic or glass pipettes for measuring small amounts of liquid.
Large graduated syringe for filling vessels with media (30ml at least).

Measuring is an important part of accuracy.  Use a protocol for measuring so that you get used to and get faster.  I prefer using small squares of wax paper for most small measurements.  I tare the scale to zero the weight then add the powdered substance.  Clean everything between uses.  Accuracy and consistency is important to success.

 

Small Hand Tools

Forceps to grasp and move explant material into vessels.
Scalpel with plastic handle preferably.
Tweezers for fine work with explants and plantlets.

There are a few small hand tools you will need.  They are all readily available from pharmacies and similar outlets.  Scalpels can be ordered online with replacement blades.  Get tools with the fewest surface textures to make sterilization easier.  Medical tools are well suited for plant tissue culture.  Long tweezers will be your most used tool.

 

PH Meter

7.0 Testing solution to calibrate the meter.
4.0 Testing solution to calibrate the meter.
Cleaning/storage solution for the meter.

PH meters should be cleaned and calibrated before each testing.  Clean them well after they have been used to.  Wash them in tap water and rinse them in tap water.  Do not mix in your agar before testing your pH in media or it may gum up the pH tester.  The cheapest pH meters will work but the more expensive ones seem to be much more accurate.

 

Common General Supplies

Baking Soda for lowering the PH of media.
White vinegar for raising the PH of media.
Spray mist bottles for bleach solution, vinegar/hydrogen peroxide solution, etc.
Plastic cling wrap to protect vessels from contamination.
Food coloring to tracking media
Mini refrigerator to store chemicals

General supplies like food coloring and plastic food wrap are easy to find.  Mist bottles can be repurposed form other products if need be but a new dedicated bottle is best.  Most don’t need to be name brand or even new.

Cooking Utensils

Pot with a lid to heat media.
Plastic ladle for stirring and dispensing.

Making media requires that it is heated properly.  Decent pots can be found with lids at many thrift stores.  Whatever you use do not use your household cookware for cooking food if you have used it for media.  Lids are a must to prevent excessive evaporation.  Don’t mix your kitchen wares with your plant tissue culture items.

 

Media Supplies

PGRs. BAP, IBA, NAA, Kinetin, 2iP
PPM (Plant Preservative Mixture)
Agar Powder
Inositol
Thiamin
Vitamin C
Murashige and Skoog Media

Ingredients for plant tissue culture media are available at most pharmacies if you want to make a DIY media.  Recently many plant tissue culture supply houses sell to hobbyists.  Otherwise everything you need you can get from a pharmacy and mix your own.

Hobby Level Plant Tissue Culture Can be Done Well

Nearly everything you need to perform plant tissue culture at the hobbyist level and be very successful is available rather easily.  Substituting a pressure cooker for an autoclave is an example of working around constraints.  There are many ways to complete tasks using other materials.  Some are cheaper, some achieve slightly fewer returns, but they work.  Some even have a better return on investment. If you run into a problem step back and think about another way to approach it.  It can be done and you can do it.

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