Laboratory Design and Layout Chapter 4. Planning of tissue culture labs Importance of asepsis Old...
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Transcript of Laboratory Design and Layout Chapter 4. Planning of tissue culture labs Importance of asepsis Old...
Planning of tissue culture labsPlanning of tissue culture labs
Importance of asepsisImportance of asepsis
Old labs – modifying ventilation and air-Old labs – modifying ventilation and air-conditioning, structural modifications will be conditioning, structural modifications will be expensiveexpensive
New labs – Integrative and innovative designNew labs – Integrative and innovative design
- Facilities may be positioned for ergonomic and - Facilities may be positioned for ergonomic and energy-saving reasonsenergy-saving reasons
Planning of tissue culture labsPlanning of tissue culture labs
Ventilation – Pressure balance and Ventilation – Pressure balance and Laminar Flow HoodsLaminar Flow Hoods
- Positive pressurePositive pressure – Avoid influx of – Avoid influx of contaminated air into surroundings of labcontaminated air into surroundings of lab
- Negative pressureNegative pressure – Usage of human material – Usage of human material- Positive-pressure buffer zone – Preparation Positive-pressure buffer zone – Preparation
area and microscopearea and microscope- Laminar Flow Hoods – Laminar Flow Hoods – Positions ofPositions of air inlets air inlets
and extractsand extracts
Planning of tissue culture labsPlanning of tissue culture labs
AccommodationAccommodation
- How many people will work each week?How many people will work each week?
- What kinds of culture will they perform?What kinds of culture will they perform?
- Area required to handle bioreactorsArea required to handle bioreactors
- 12 laminar hoods/50 people12 laminar hoods/50 people
Planning of tissue culture labsPlanning of tissue culture labs
Space –Space –- Largest area – culture operation accommodating laminar-flow Largest area – culture operation accommodating laminar-flow
hoods, cell counters, centrifuges, incubators, microscopes, and hoods, cell counters, centrifuges, incubators, microscopes, and arrangements for media, stocks of reagents, glassware and arrangements for media, stocks of reagents, glassware and plastics etc.plastics etc.
- Area for wash up, preparation and sterilizationArea for wash up, preparation and sterilization
- Area for storageArea for storage
- Area for incubationArea for incubation
Planning of tissue culture labsPlanning of tissue culture labs
Aseptic areaAseptic area
- Animal facilityAnimal facility is accessible but not is accessible but not contiguous to tissue culture labscontiguous to tissue culture labs
- WindowsWindows – high heat generation, UV influx – high heat generation, UV influx and incursion of microorganismsand incursion of microorganisms
Planning of tissue culture labsPlanning of tissue culture labs
HoodsHoods – –
- Space between hoods should be - Space between hoods should be 500 mm (2 ft)500 mm (2 ft)
- - Motion movementMotion movement
- Minimize interferenceMinimize interference in airflow between in airflow between hoodshoods
- Space filled with Space filled with removable cart or trolleyremovable cart or trolley
Planning of tissue culture labsPlanning of tissue culture labs
IncubationIncubation – –
- Large number of flasks or large volume flasks Large number of flasks or large volume flasks (sealed) incubated in (sealed) incubated in hot roomshot rooms
- Open plates and dishes incubated in Open plates and dishes incubated in humid humid Co2 incubatorCo2 incubator
Planning of tissue culture labsPlanning of tissue culture labs
Preparation areaPreparation area – – - Facilities for washing up and for sterilization Facilities for washing up and for sterilization
should be locatedshould be located
- i) close to aseptic area that they servicei) close to aseptic area that they service
- ii) outside wall to allow for heat extractions ii) outside wall to allow for heat extractions from ovens and steam vents from autoclavesfrom ovens and steam vents from autoclaves
Planning of tissue culture labsPlanning of tissue culture labs
Servicing aseptic areasServicing aseptic areas – –
- - Elevator or ramp sufficeElevator or ramp suffice
- Maximum load capacity of ramp without any - Maximum load capacity of ramp without any mechanical helpmechanical help
StorageStorage – –
- Space required for disposable plasticsSpace required for disposable plastics
- Cell line work with liquid nitrogenCell line work with liquid nitrogen
Planning of tissue culture labsPlanning of tissue culture labs
RenovationsRenovations – Choose location carefully – Choose location carefully
AccessAccess – Doorways are wide and high to allow – Doorways are wide and high to allow for installationsfor installations
QuarantineQuarantine - -
- Screening for Screening for MycoplasmaMycoplasma in new cell lines in new cell lines and biopsiesand biopsies
- Containment Containment of biopsies and cell lines posing of biopsies and cell lines posing biohazardbiohazard
Layout of aseptic room or suiteLayout of aseptic room or suite
Six main functionsSix main functions – sterile handling, – sterile handling, incubation, preparation, washup, sterilization incubation, preparation, washup, sterilization and storageand storageLocation for sterile handling should be at Location for sterile handling should be at one one end of roomend of room
- Wash up and sterilization facilities Wash up and sterilization facilities at other at other endend
- In-betweenIn-between - Preparation, Storage and - Preparation, Storage and Incubation Incubation
4.3.1 Sterile handling area4.3.1 Sterile handling area
Sterile area restrictedSterile area restricted
No traffic or disturbanceNo traffic or disturbance
Absence of laminar hoods – use separate room Absence of laminar hoods – use separate room or cubicleor cubicle
- Plastic laminate topped bench - Plastic laminate topped bench
- Freestanding or sealed to wall - Freestanding or sealed to wall
4.3.2 Laminar Hood4.3.2 Laminar Hood
Less expensive than sterile roomsLess expensive than sterile roomsHoodsHoods – Operator’s arms enter – Operator’s arms enterLaminar flow wall or ceiling unitsLaminar flow wall or ceiling units – – Operator is part of work areaOperator is part of work area
- Wear sterile caps and gownsWear sterile caps and gowns- Legroom with space for pumps, aspiratorsLegroom with space for pumps, aspirators- Chairs with adjustable seat and heightChairs with adjustable seat and height- Cart or trolley or folding flap (300-500mm)Cart or trolley or folding flap (300-500mm)
4.3.3 Quarantine and Containment4.3.3 Quarantine and Containment
Separate room with door or lockSeparate room with door or lock
Separate incubators, freezer, refrigerator, centrifuge, Separate incubators, freezer, refrigerator, centrifuge, supplies and disposalsupplies and disposal
Newly imported cell lines or biopsiesNewly imported cell lines or biopsies
Act as Act as Level II containmentLevel II containment – biohazard cabinet or – biohazard cabinet or pathogen trappathogen trap
4.3.4 Service bench4.3.4 Service bench
Close to sterile handling areaClose to sterile handling area
Drawer units below and open shelves aboveDrawer units below and open shelves above
Store sterile glasswares, plastics, pipettes, Store sterile glasswares, plastics, pipettes, screw caps, syringes etcscrew caps, syringes etc
4.4 Incubation4.4 Incubation
IncubatorsIncubators
Less expensiveLess expensive
Lose more heat and Lose more heat and slow to recoverslow to recover
Hot roomHot room
Less expensive than two Less expensive than two incubatorsincubators
Insulated to prevent Insulated to prevent cold spotscold spots
Avoid wooden Avoid wooden furnishingsfurnishings
Stainless steel bench or Stainless steel bench or plastic laminateplastic laminate
Heat generation in hot roomHeat generation in hot room
Heater Heater – Fan heater provides heat, run – Fan heater provides heat, run continuouslycontinuously
Air circulationAir circulation – Fan positioned on opposite – Fan positioned on opposite side of room of fan heaterside of room of fan heater
ThermostatsThermostats – Two separate heaters with – Two separate heaters with their thermostatstheir thermostats
OverheatingOverheating – increase of room temperature, – increase of room temperature, heat produced from apparatus etcheat produced from apparatus etc
4.5 Preparation area4.5 Preparation area
4.5.1 4.5.1 Area for media preparationArea for media preparation - -
- - Accommodate balances, pH meter, osmometer Accommodate balances, pH meter, osmometer etcetc
- Preparation and sterilization in hoods- Preparation and sterilization in hoods
4.5.2 4.5.2 Wash up and sterilization facilitiesWash up and sterilization facilities outside tissue culture lab outside tissue culture lab
- Space for drying glassware, sorting pipettes, - Space for drying glassware, sorting pipettes, packaging and sealing packs for sterilizationpackaging and sealing packs for sterilization
4.5 Preparation area4.5 Preparation area
4.5 Wash up4.5 Wash up
Raised edge – to avoid spillageRaised edge – to avoid spillage
Four taps Four taps
Deionized water provisionDeionized water provision
4.5 Preparation area4.5 Preparation area
4.5.3 Storage4.5.3 Storage
Sterile liquids – 25, 4 or -20 or – 70 degreeSterile liquids – 25, 4 or -20 or – 70 degree
Sterile and non sterile glasswareSterile and non sterile glassware
Sterile disposable plasticsSterile disposable plastics
Screw caps, stoppersScrew caps, stoppers
FiltersFilters
Gloves and disposal bagsGloves and disposal bags
4.5.3 Storage4.5.3 Storage
Liquid Nitrogen – Liquid Nitrogen – Dewars (25-50 L)Dewars (25-50 L)
Large storage vessel Large storage vessel (100-150 L)(100-150 L) on trolley on trolley
Storage tanks Storage tanks (500 – 1000 L)(500 – 1000 L)
Safety – Safety – Ventilation + oxygen alarm + replace Ventilation + oxygen alarm + replace air in roomair in room
Co2 storage unitsCo2 storage units –tethered to wall or bench –tethered to wall or bench in a rackin a rack
This project is funded by a grant awarded under the President’s Community Based Job Training Grant as 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-15-162-06-60). implemented by the U.S. Department of Labor’s Employment and Training Administration (CB-15-162-06-60). NCC is an equal opportunity employer and does not discriminate on the following basis: 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, against any individual in the United States, on the basis of race, color, religion, sex, national origin, age disability, political affiliation or belief; andpolitical affiliation or belief; and
against any beneficiary of programs financially assisted under Title I of the Workforce Investment Act of 1998 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 (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.the United States, or his or her participation in any WIA Title I-financially assisted program or activity.
DisclaimerThis 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.