Laboratory Design and Layout

Chapter 4

Planning of tissue culture labs

Importance of asepsis Old labs modifying ventilation and airconditioning, 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 Operators 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

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

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

This project is funded by a grant awarded under the Presidents Community Based Job Training Grant as implemented by the U.S. Department of Labors Employment and Training Administration (CB -15-162-06-60). NCC is an equal opportunity employer and does not discriminate on the following basis: against any individual in the United States, on the basis of race, color, religion, sex, national origin, age disability, political affiliation or belief; and against any beneficiary of programs financially assisted under Title I of the Workforce Investment Act of 1998 (WIA), on the basis of the beneficiarys citizenship/status as a lawfully admitted immigrant authorized to work in the United States, or his or her participation in any WIA Title I-financially assisted program or activity.

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