The document outlines the key considerations for designing an effective plant tissue culture laboratory. It discusses that the laboratory should have separate areas for washing and storage, media preparation, aseptic transfer, and primary growth. The aseptic transfer area and primary growth room require stringent cleanliness, with HEPA filters, UV lights, and laminar flow hoods. Other areas require equipment for media preparation, sterilization, and storage. The primary growth room needs temperature, humidity, and lighting controls to suit the plants being cultured. Careful planning of traffic flow and separation of clean and dirty areas is important for success.

1. Designing a Plant Tissue
Culture Laboratory
Dr Fatimah Kayat

2. Outline
 Introduction
 General laboratory design
 Washing & Storage
 Media Preparation
 Instrumentation
 Aseptic transfer area
 Primary growth room

3. INTRODUCTION
 Certain elements are essential for a
successful operation
 The correct and strategic design helps to
achieve a high standard work.
 Careful planning is an important step
considering the size and location.
 A small lab should be set up first until the
proper techniques and markets are
developed.
 STRATEGIC PLANNING

5.  Good locations
Isolation from traffic.
No contamination from adjacent rooms.
 Access to water & electricity supply
 Connection to a septic system or sanitary
sewer
CONSIDER

6. CONSIDER
 Water service
 Air conditioning
 maintain constant temperature
 Electrical service capacity
 equipment, lights and future expansion
 An emergency generator
 to operate essential equipment during
power breakdown

7. PRIORITY
 Cleanliness
 No contamination.
 Easy to wash walls and floors.
 High efficiency particulate air (HEPA) filters.
 An enclosed entrance, sticky mats, no shoes.

8.  Traffic pattern and work flow
 The cleanest rooms or areas
 Aseptic transfer area
 Primary growth room
 Could not enter directly from outside.
 Should be enclosed with doors leading to each other
 Media preparation area, washing area, storage
area should be outside these rooms.
 Media preparation area should lead to the
sterilization area
TO CONSIDER

9.  Washing area located near sterilization and media
preparation area.
 should be equipped with at least one large sink.
 The pipes can be PVC to resist damage from acids
and alkaline
 Hot/cold water sources
 Water distillation and/or deionization devices
 Ovens or hot air cabinets (75 oC) close to the washing
and storage area
WASHING & STORAGE AREA

10.  Storage area should be close to the washing
area for storage
 Need to be accessible to the media preparation
area
 Dust-proof cabinets can be used in the storage
area
WASHING & STORAGE AREA

11.  Glassware
Good quality of borosilicate glass
Highly resistant to autoclaving, breakage and
scratch proof
E.g: Pyrex, Corning, Kimble, Schott

12.  Water source and storage area should be
convenient to the media preparation area.
 A variety of equipment for media preparation are
generally located in the media preparation area.
MEDIA PREPARATION &
STERILIZATION AREA

13. BASICS EQUIPMENT
 Refrigerator/freezer
 to store chemicals and stock solutions
(usually in media preparation area)
 High quality water
 Purchasing bottled water is not economical for
mass production
 Distilled water, double distilled water devices
(usually in washing area)

14.  A triple beam balance
 Useful for large amounts over 10 grams
 Can measure down to 2 mg is essential.
 Balances
 a microbalance
 a less sensitive top loading balance
BASICS EQUIPMENT

15.  Hot plate/stirrer
 At least one hot plate with an automatic
stirrer is needed to make semi-solid media.
BASICS EQUIPMENT

16.  pH meter
 to measure media pH.
 Some laboratories use pH indicator paper.
BASICS EQUIPMENT

17.  Aspirator or vacuum pump
 to filter sterilization of chemicals
 also used to disinfest plant material
Other Equipment

18.  Autoclave or Pressure Cooker
 vital part of a laboratory
 to sterile media, apparatus, used vessel
 Self generating steam autoclaves are more
dependable and faster to operate.
BASICS EQUIPMENT

19.  Optional equipment
Microwave oven are convenient for defrosting
frozen stocks and heating agar media
Dissecting microscopes are useful for
dissecting floral and shoot apices, and
observing plant culture growth
Labwashers/dishwashers are useful for
washing and drying glassware (mass
production)
OTHER EQUIPMENT

20.  Laminar flow hood
 For sterile working procedure
With High Efficiency Particulate Air (HEPA)
filter – filter the air to high degree
 With UV germicidal lamp – sterile the shell and
content when not in use
Horizontal laminar flow unit---An enclosed work area that has sterile
air moving across it. The air moves with uniform velocity along
parallel flow lines. Room air is pulled into the unit and forced through
a HEPA (High Energy Particulate Air) filter, which removes particles
0.3 μm and larger.
BASICS EQUIPMENT

21. LAMINAR FLOW HOOD

24.  Incubator
 To incubate the culture
 Controlled conditions (temperature, light,
humidity)
OTHER EQUIPMENT

25.  Need to be as clean as possible
 Preferably separated from other rooms
 to decreases spore circulation and allows personnel to
leave shoes outside the room.
 Laboratory shoes and lab coats should be worn
 Aseptic working area.
 Ultraviolet (UV) lights are sometimes installed to
disinfect the room (only switched on when people
and plant material are not in the room)
 Safety switches can be installed to shut off the UV
lights.
 air filters are sometimes installed to provide clean air
ASEPTIC TRANSFER AREA

26.  Smooth surfaces inside the aseptic transfer
area
 to minimize the amount of dust and easy to wipe.
 Several electric outlets are to be installed to
accommodate
 Flow hoods,
 Hot bead sterilizer
 Microscopes.
ASEPTIC TRANSFER AREA

27. ASEPTIC TRANSFER AREA –
commercial production

28. PRIMARY GROWTH
ROOM
 Should consider:
 temperature
 relative humidity
 lighting units and shelves.
 Varies depending on
 the size of the growth room
 its location
 the type of plants grown

29.  Temperature is the primary concern
 It affects decisions on lights, relative humidity, and
shelving.
 Generally, temperatures are kept at 24 ±1 oC.
 Cooling the room is usually a greater problem
than heating;
 cooler T can be obtained by installing heat pumps,
air conditioners or exhaust fans.
 Small fans can be placed at the ends of shelves
 to increase air flow and decrease heat accumulation.
PRIMARY GROWTH ROOM

30. PRIMARY GROWTH ROOM
 Shelving within primary growth rooms can vary
depending upon the situation and the plants
grown.
 Wood is inexpensive and easy-to-build shelves.
 It could be particleboard or plywood and painted
white to reflect the room's light.
 Expanded metal is more expensive but provides
better air circulation.

31.  Some plant cultures can /need to be kept in
complete darkness
 Most culture rooms are under light condition
 Depending on
 plant species
 developmental stage
 Automatic timers
 help to maintain desired photoperiods
 Reflectors can replaced bulbs
 reduce cost
 Heat generated by lights may cause
condensation and temperature problems.
PRIMARY GROWTH ROOM

32.  Relative humidity (RH) is difficult to control
inside growing vessels
 fluctuations in the culture room may have a
deleterious effect.
 Cultures can dry out if the room's RH is less
than 50%;
 humidifiers can be used to correct this problem.
 If the RH becomes too high, a dehumidifier
is recommended.
PRIMARY GROWTH ROOM

33. PRIMARY GROWTH ROOM

34. PRIMARY GROWTH ROOM

35. PRIMARY GROWTH ROOM

36. Any questions?