Laboratory Design and Layout Chapter 4

Planning of tissue culture labs Importance of asepsis Old labs – modifying ventilation and air-conditioning, structural modifications will be expensive New labs – Integrative and innovative design - Facilities may be positioned for ergonomic and energy-saving reasons

Planning of tissue culture labs Ventilation – Pressure balance and Laminar Flow Hoods Positive pressure – Avoid influx of contaminated air into surroundings of lab Negative pressure – Usage of human material Positive-pressure buffer zone – Preparation area and microscope Laminar Flow Hoods – Positions of air inlets and extracts

Planning of tissue culture labs Accommodation How many people will work each week? What kinds of culture will they perform? Area required to handle bioreactors 12 laminar hoods/50 people

Planning of tissue culture labs Space – Largest area – culture operation accommodating laminar-flow hoods, cell counters, centrifuges, incubators, microscopes, and arrangements for media, stocks of reagents, glassware and plastics etc. Area for wash up, preparation and sterilization Area for storage Area for incubation

Planning of tissue culture labs Aseptic area Animal facility is accessible but not contiguous to tissue culture labs Windows – high heat generation, UV influx and incursion of microorganisms

Planning of tissue culture labs Hoods – - Space between hoods should be 500 mm (2 ft) - Motion movement Minimize interference in airflow between hoods Space filled with removable cart or trolley

Planning of tissue culture labs Incubation – Large number of flasks or large volume flasks (sealed) incubated in hot rooms Open plates and dishes incubated in humid Co2 incubator

Planning of tissue culture labs Preparation area – Facilities for washing up and for sterilization should be located i) close to aseptic area that they service ii) outside wall to allow for heat extractions from ovens and steam vents from autoclaves

Planning of tissue culture labs Servicing aseptic areas – - Elevator or ramp suffice - Maximum load capacity of ramp without any mechanical help Storage – Space required for disposable plastics Cell line work with liquid nitrogen

Planning of tissue culture labs Renovations – Choose location carefully Access – Doorways are wide and high to allow for installations Quarantine - Screening for Mycoplasma in new cell lines and biopsies Containment of biopsies and cell lines posing biohazard

Layout of aseptic room or suite Six main functions – sterile handling, incubation, preparation, washup, sterilization and storage Location for sterile handling should be at one end of room Wash up and sterilization facilities at other end In-between - Preparation, Storage and Incubation

4.3.1 Sterile handling area Sterile area restricted No traffic or disturbance Absence of laminar hoods – use separate room or cubicle - Plastic laminate topped bench - Freestanding or sealed to wall

4.3.2 Laminar Hood Less expensive than sterile rooms Hoods – Operator’s arms enter Laminar flow wall or ceiling units – Operator is part of work area Wear sterile caps and gowns Legroom with space for pumps, aspirators Chairs with adjustable seat and height Cart or trolley or folding flap (300-500mm)

4.3.3 Quarantine and Containment Separate room with door or lock Separate incubators, freezer, refrigerator, centrifuge, supplies and disposal Newly imported cell lines or biopsies Act as Level II containment – biohazard cabinet or pathogen trap

4.3.4 Service bench Close to sterile handling area Drawer units below and open shelves above Store sterile glasswares, plastics, pipettes, screw caps, syringes etc

4.4 Incubation Incubators Less expensive Lose more heat and slow to recover Hot room Less expensive than two incubators Insulated to prevent cold spots Avoid wooden furnishings Stainless steel bench or plastic laminate

Heat generation in hot room Heater – Fan heater provides heat, run continuously Air circulation – Fan positioned on opposite side of room of fan heater Thermostats – Two separate heaters with their thermostats Overheating – increase of room temperature, heat produced from apparatus etc Thermometer in room

4.5 Preparation area 4.5.1 Area for media preparation - - Accommodate balances, pH meter, osmometer etc - Preparation and sterilization in hoods 4.5.2 Wash up and sterilization facilities outside tissue culture lab - Space for drying glassware, sorting pipettes, packaging and sealing packs for sterilization To prevent dissipation of heat. Space for a pipette washer and dryer. Sinks deep enough (450 mm, 18 inches) allow for manual washing and rinsing. Tall persons will have to bend so height of sink should be high rather than low. A short person can stand on a height to reach the sink

4.5 Preparation area 4.5 Wash up Raised edge – to avoid spillage Four taps Deionized water provision

4.5 Preparation area 4.5.3 Storage Sterile liquids – 25, 4 or -20 or – 70 degree Sterile and non sterile glassware Sterile disposable plastics Screw caps, stoppers Filters Gloves and disposal bags

4.5.3 Storage Liquid Nitrogen – Dewars (25-50 L) Large storage vessel (100-150 L) on trolley Storage tanks (500 – 1000 L) Safety – Ventilation + oxygen alarm + replace air in room Co2 storage units –tethered to wall or bench in a rack Two openings of storage units. Cold storage as a room or refrigerator

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