1. Laboratory Design and Layout
Chapter 4

2. 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

3. 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

4. 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

5. 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

6. 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

7. 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

8. 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

9. 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

10. 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

11. 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

12. 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

13. 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

14. 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 ( mm)

15. 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

16. 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

17. 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

18. 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

19. 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

20. 4.5 Preparation area 4.5 Wash up Raised edge – to avoid spillage
Four taps
Deionized water provision

21. 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

22. 4.5.3 Storage Liquid Nitrogen – Dewars (25-50 L)
Large storage vessel ( 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

23. This project is funded by a grant awarded under the President’s Community Based Job Training Grant as implemented by the U.S. Department of Labor’s Employment and Training Administration (CB ). 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 beneficiary’s 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.

24. Disclaimer
This workforce solution was funded by a grant awarded under the President’s Community-Based Job Training Grants as implemented by the U.S. Department of Labor’s Employment and Training Administration.  The solution was created by the grantee and does not necessarily reflect the official position of the U.S. Department of Labor.  The Department of Labor makes no guarantees, warranties, or assurances of any kind, express or implied, with respect to such information, including any information on linked sites and including, but not limited to, accuracy of the information or its completeness, timeliness, usefulness, adequacy, continued availability, or ownership.  This solution is copyrighted by the institution that created it.  Internal use by an organization and/or personal use by an individual for non-commercial purposes is permissible.  All other uses require the prior authorization of the copyright owner.