How to set up a tissue culture laboratory?

How to set up a tissue culture laboratory?

Plant tissue culture has been a booming field of science over the last few decades. Nowadays, its knowledge is so readily available that anyone with genuine interest can do it. In the long run, it can generate enormous financial returns. However, the first step is to set up a tissue culture laboratory!

A lot of you might be curious about how you can set up your own tissue culture laboratory. What are the requirements? What equipment do you need to have? Where you need to be careful while planning a setup? In this article we will briefly try to solve this mystery!

Tissue culture plants are quite useful because they are uniform and you can obtain high quality plants in relatively short duration. Different businesses can profit from plant tissue culture in innovative ways. And if you want to know more about how plant tissue culture can help you generate profits, then do have a look at our blog on "8 ways plant tissue culture generates profit".

Also, if you are thinking of setting up a tissue culture facility for research or commercial purposes on a small to large-scale production levels, you can always contact our wonderful team for consultation and guidance! We specialize in designing and setting up plant tissue culture and seed breeding laboratories. Even for specific laboratory needs, questions or additional support, we are there to help you out!

For now, let us talk about a basic commercial laboratory set up.

Lab area requirements

In order to set up your plant tissue culture laboratory, it is important that you carefully choose the location. The sterile environment of a tissue culture lab is prone to contamination through air vents. You should try not to built it close to a facility which deals with micro-organisms (bacteria, fungi and so on). You should also try that it should not be in the close vicinity of other laboratories which handle insects. Another important point is that the lab should be away from storage places which store seeds and old plant material. However, if you cannot avoid these then you need to take extra precautions to keep the lab free from contamination.

For any tissue culture lab, there are certain basic requirements or facilities that you need to install for efficient functioning and experimentation. These are:

  • Quarantine area
  • Washing area
  • Media preparation area
  • Aseptic transfer area
  • Culture/grow rooms
  • Data collection area

Let us dive a bit deeper into these areas of a plant tissue culture lab!

Quarantine area

This area is essential to prevent contamination coming from us - humans! It can be present only in the beginning of the lab or both in the beginning and exit of a lab. This area is primarily for you to change dresses and shoes. There should be separate shoes and lab coats for you to use in the lab.

Washing area

You need this area to to wash the explants or other plant materials. You also need this to clean all your beakers, cylinders and flasks which you will use for preparing media or measuring ingredients. This area consists of large sinks and draining boards. This area also has access to distilled water with space for drying. Thus it facilitates the preliminary cleaning to kill the microorganisms present on the surface.

Media preparation area

You need this area for preparing culture media recipes and for storage of chemicals and glassware. You can use a large area for media preparation with benches. This area also has hot plates, pH meters, weighing balances and water baths. You can also choose to have a separate area for the media dispensing unit which is connected to your media preparator or autoclave present in the preparation area. The important thing to know is the equipment we need for media preparation which are:

  • Magnetic stirrer
  • Analytical and top-loading balances
  • pH meter
  • Refrigerator with freezer or a separate freezer for storing stock solutions, plant hormones and other chemicals
  • Water purification system along with the space to store the purified water
  • Equipment for sterilization of media, glassware, and instruments (pressure cooker or autoclaves for small lab setup, automated media preparators for research, medium and large scale commercial tissue culture setups)

Media preparation is a crucial step, and you need to develop a system to have a step-by-step routine for preparing media and careful measurement of chemicals.

DID YOU KNOW?

Our ‘Premium’ 70 liter media preparator model is the fastest and most reliable for its size in the market with a very short sterilization cycle time of just 30 minutes on a stand alone basis!

Transfer area

In ideal conditions, a separate room should be made next to the media preparation area or next to the area for the media dispensing unit. However, you need this area to have complete sterile conditions in order to transfer explants to media containing culture vessels. So the activities for this room are:

  • Transfer of explants to growth media;
  • For sub culturing;
  • For multiplication transfers; and
  • For transferring multiplied shoots to rooting media.

One of the most important key points for this area is that it should always have positive pressure airflow. You need positive pressure airflow to make sure that air can leave the room without circulating back in.

Another important thing for you is to construct this area in a way that all the surfaces stay free from dust and microorganisms. You need to always clean this area thoroughly.

For making transfers in this area, you need to install laminar flow cabinets as well. This is to to ensure constant flow of bacteria-free filtered air and you do sub culturing and other transfers within these cabinets. You need this also to prevent non-filtered air and particulate matter from settling down on the surface while making transfers. So this works by forcing air into the laminar unit through a dust filter and then passing it through a HEPA filter (high-efficiency particulate air filter). This is known as a flow unit and you have two types:

  • Vertical flow unit: the air from HEPA filter is directed downwards.
  • Horizontal flow unit: the air from HEPA filter is directed outwards.

Culture/grow room

After transferring explants and shoots to the media, you need to store them in ideal conditions for efficient growth and development. This means that you need to provide controlled temperature, humidity, air circulation, light quality, and duration of light. You should know that an automated climate and air filtration system works best for regulating these conditions.

These environmental factors are important for the plant cells to grow and differentiate into healthy shoots and roots at later stages. These specific conditions differ with each plant species. Hence, you need to be careful with providing the necessary growth environment for optimum development of tissue culture plantlets.

We will discuss these factors in detail in another article!

Data collection area

Developing tissue culture protocols for different plant species and varieties involves a lot of trial-and-error experiments. You will be experimenting with culture media recipes for different growth stages and also with different growth conditions in the culture/grow room. Thus, you need this area to have efficient record keeping practices for different kind of tissue culture experiments. Hence, getting yourself a desktop computer for writing up reports and analyzing the data will be a great idea!

So with these areas in place, a tissue culture laboratory can be efficiently set up with productive workflow. If you are setting up a lab for commercial production, then you also need to focus not just on product workflow, but also on people workflow. Apart from this, research and development of tissue culture protocols for each plant species also play a key role in the success of your plant tissue culture work and production.

We hope that this brief description will be useful for many of you! Keep checking this space for informative articles!

By Nancy Bhatia | 26-May-2021

References

  1. Robert H. Smith (2013). Plant Tissue Culture- Techniques and Experiments. Elsevier Inc.
  2. https://link.springer.com/chapter/10.1007/978-0-585-27571-0_2
  3. https://biotopeone.com/7-setting-up-a-home-plant-tissue-culture-lab/
  4. https://www-pub.iaea.org/MTCD/Publications/PDF/te_1384_web.pdf