Laboratory Design and LayoutChapter 4

Planning of tissue culture labsImportance of asepsisOld labs modifying ventilation and air-conditioning, structural modifications will be expensiveNew labs Integrative and innovative design- Facilities may be positioned for ergonomic and energy-saving reasons

Planning of tissue culture labsVentilation Pressure balance and Laminar Flow HoodsPositive pressure Avoid influx of contaminated air into surroundings of labNegative pressure Usage of human materialPositive-pressure buffer zone Preparation area and microscopeLaminar Flow Hoods Positions of air inlets and extracts

Planning of tissue culture labsAccommodation

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 labsSpace 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 labsAseptic 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 labsHoods - Space between hoods should be 500 mm (2 ft)- Motion movementMinimize interference in airflow between hoodsSpace filled with removable cart or trolley

Planning of tissue culture labsIncubation

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 labsPreparation area Facilities for washing up and for sterilization should be locatedi) close to aseptic area that they serviceii) outside wall to allow for heat extractions from ovens and steam vents from autoclaves

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

Planning of tissue culture labsRenovations Choose location carefullyAccess Doorways are wide and high to allow for installationsQuarantine - Screening for Mycoplasma in new cell lines and biopsiesContainment of biopsies and cell lines posing biohazard

Layout of aseptic room or suiteSix main functions sterile handling, incubation, preparation, washup, sterilization and storageLocation for sterile handling should be at one end of roomWash up and sterilization facilities at other endIn-between - Preparation, Storage and Incubation

4.3.1 Sterile handling areaSterile area restrictedNo traffic or disturbanceAbsence of laminar hoods use separate room or cubicle - Plastic laminate topped bench - Freestanding or sealed to wall

4.3.2 Laminar HoodLess expensive than sterile roomsHoods Operators arms enterLaminar flow wall or ceiling units Operator is part of work areaWear sterile caps and gownsLegroom with space for pumps, aspiratorsChairs with adjustable seat and heightCart or trolley or folding flap (300-500mm)

4.3.3 Quarantine and ContainmentSeparate 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 IncubationIncubatorsLess expensiveLose more heat and slow to recoverHot roomLess expensive than two incubatorsInsulated to prevent cold spotsAvoid wooden furnishingsStainless steel bench or plastic laminate

Heat generation in hot roomHeater Fan heater provides heat, run continuouslyAir circulation Fan positioned on opposite side of room of fan heaterThermostats Two separate heaters with their thermostatsOverheating increase of room temperature, heat produced from apparatus etc

4.5 Preparation area4.5.1 Area for media preparation - - Accommodate balances, pH meter, osmometer etc- Preparation and sterilization in hoods4.5.2 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 spillageFour taps Deionized water provision

4.5 Preparation area4.5.3 StorageSterile liquids 25, 4 or -20 or 70 degreeSterile and non sterile glasswareSterile disposable plasticsScrew caps, stoppersFiltersGloves and disposal bags

4.5.3 StorageLiquid Nitrogen Dewars (25-50 L)Large storage vessel (100-150 L) on trolleyStorage tanks (500 1000 L)Safety Ventilation + oxygen alarm + replace air in roomCo2 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; andagainst 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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*Thermometer in room*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*Two openings of storage units. Cold storage as a room or refrigerator