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Autoclave Training Guide. York University Department of Occupational Health and Safety Kinsmen Building, 8 The Chimneystack Road Telephone : 416-736-5491 Revised: January 2012. Autoclaving Training Outline. Autoclave Overview Autoclave Hazards

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autoclave training guide

Autoclave Training Guide

York University

Department of Occupational Health and Safety

Kinsmen Building, 8 The Chimneystack Road

Telephone : 416-736-5491

Revised: January 2012

autoclaving training outline
Autoclaving Training Outline
  • Autoclave Overview
  • Autoclave Hazards
  • What can be Autoclaved
  • How to Autoclave
    • Cycle selection
    • Proper containers
    • Proper loading/unloading
  • Autoclave Performance Indicators
steam sterilization
Steam Sterilization
  • Pressurized device that uses heat, steam and pressure to achieve sterilization or decontamination
autoclave overview
Autoclave Overview
  • Typically operated at 121°C, 15psi, for 15-45 minutes.
  • Allows the heating of liquids above boiling point.
  • Uses moist heat (steam) to increase efficiency of sterilization.
  • Heat is used to coagulate proteins, which destroys microorganisms and any potential biohazard.
autoclave hazards
Autoclave Hazards
  • Tremendous pressure from steam in chamber provides explosive potential.
  • High temperatures creates potential for burns and scalding.
  • Potential exposure to hazardous fumes
  • Inadequate decontamination allows for the potential of biological hazards to contaminate personnel and the environment.
what you can autoclave
What you CAN autoclave

Biological waste that can be autoclaved:

  • Microbial stocks and cultures
  • Items contaminated with such waste: petri dishes, eppendorf tips, pipettes, gloves, paper towel

Autoclaving is also used for:

  • Items for sterilizations such as; glassware media, aqueous solutions
what you can t autoclave
What you CAN’T Autoclave
  • DO NOT Autoclave:
    • BIOMEDICAL WASTE (except Microbiological)
      • Including human anatomical or blood waste, animal anatomical or blood waste, cytotoxic waste, or any waste in contact with these waste products (including sharps waste).
      • This includes anything contaminated with a toxic, volatile, corrosive, or mutagenic chemical
      • (e.g.) bleach, formalin, glutaraldehyde, ethidium bromide
      • Check MSDS beforehand
autoclaves used to treat biohazardous waste at york
Autoclaves used to treat biohazardous waste at York
  • Lumbers Room 120A
  • Farquharson 227A
  • Life Sciences Building
    • 3rd Floor
how to autoclave
How to Autoclave
  • Preparing your items for autoclaving
  • Loading the autoclave
  • Choosing the cycle settings
  • Unloading the autoclave
  • Aborting Autoclave
autoclaving preparing your items
Autoclaving: Preparing your items
  • Sign into log book
    • Keeps track of autoclave use for maintenance records
  • Use personal protective equipment!
    • Eye protection
    • Heat resistant gloves
    • Lab coat
autoclaving preparing your items1
Autoclaving: Preparing your items
  • Use a primary container
    • Container comes into direct contact with the contaminated or non-sterilized material or fluid
    • Do not fill more than 75% of holding capacity
autoclaving preparing your items2
Autoclaving: Preparing your items
  • Use a primary container (cont’d)
    • Must NOT be a tightly sealed container (might explode)
    • Primary container must permit heat (steam) penetration
      • Loosen screw caps or use self venting caps
      • Cap open containers with aluminum foil or muslin
      • If using plastic waste bags, leave a small opening

Small opening

Loosen screw caps

autoclaving preparing your items3
Autoclaving: Preparing your items
  • Use a secondary container
    • Used to contain any spills
    • The sides of the secondary container must be sufficiently high to contain any spill that may occur
    • Tray MUST be autoclave safe

For liquids add water

autoclaving preparing your items4
Autoclaving: Preparing your items
  • Use temperature sensitive tape
    • Will indicate that high temperature has been achieved
    • Will not prove that decontamination or sterilization was successful
    • Will assist in keeping track of autoclave and non autoclaved items



autoclaving loading the autoclave
Autoclaving: Loading the autoclave
  • NEVER autoclave liquid loads using a solid load cycle
    • liquids will boil over
  • Be cautious if autoclave was recently used
    • Rack may be hot
    • Use heat protective gloves
  • Make sure tray is locked on dolly
    • Rack may slide out unexpectedly if not locked
autoclaving loading the autoclave1
Autoclaving: Loading the autoclave
  • Farquharson:
    • Tighten door so that prongs are fully extended
    • If noise and/or steam escapes, abort the cycle and tighten the door more
  • Lumbers
    • Keep the door button pushed until the ready prompt is on the screen
autoclaving which cycle to use
Autoclaving: Which cycle to use
  • Autoclaves run 3 types of cycle programs
  • The type of cycle depends on what is being autoclaved:

Note: for Farquharson, use Solid/Gravity for wrapped, solid items

autoclaving choosing a cycle
Autoclaving: Choosing a Cycle
  • For Liquids:
    • 20 mins / litre of liquid, 5 mins per additional litre
  • For Solids:
    • Glassware (empty): 15 mins
    • Instruments (utensils): 30 mins
    • Biohazardous Garbage: at least 30 mins per full bag
      • Use biological test strips to optimize duration
autoclaving proper loading

Autoclaved (regular)

Autoclaved (due to being compressed & sealed)

Autoclaving: Proper loading
  • Autoclave biohazardous waste separately
  • Do not over load primary or secondary containers
    • Allow for sufficient steam penetration
    • Do not fill more than 75% to allow expansion without overflow
  • Do not pack or compress contents
autoclaving unloading autoclave
Autoclaving: Unloading autoclave
  • Use PPE
  • Wait for autoclave to state END CYCLE before opening door
  • When opening, stand away from door opening
    • Make sure no one is standing by door opening
  • Farquharson: turn door knob slowly and open door slightly to allow steam to escape
  • Lumbers: make sure your hands are not above the top vents
how to autoclave unloading autoclave
How to Autoclave – Unloading autoclave
  • Make sure rack is locked on to dolly
  • Unmark any biohazard signs that may be seen on waste bags
  • Dispose of waste in proper bins
  • Please put back heat resistant gloves for other users
  • Keep autoclave doors shut when not in use
how to autoclave unloading autoclave1
How to Autoclave – Unloading autoclave
  • Unmark any biohazard signs and words that may be seen on waste bags!
how to autoclave aborting
How to Autoclave – Aborting
  • Only qualified personnel should attempt to troubleshoot an autoclave
  • Farquharson
    • Large autoclave: push abort button
    • Small autoclave: need to manually advance through the autoclave cycle
  • Lumbers
    • Push abort button
autoclave performance indicators
Autoclave: Performance Indicators
  • How to know if autoclave is functioning correctly:
autoclave issues concerns
Autoclave Issues/Concerns?


  • Your Supervisor

Farquharson and Lumbers:

  • Jane Grant, Biology, Lumbers 130, x22657
  • Debbie Freele, Biology, Lumbers 115, x22655
  • Prof. Roger Lew, Biology, Farq 230, x66114
  • Brad Sheeller, FSE


  • Prof. Vivian Saridakis
  • Brad Sheeller, FSE

York Biosafety Officer:

  • Francis Arnaldo, DOHS, x44745
autoclave training
Autoclave Training
  • Completion of in-class autoclave training is included with the Biosafety Training session.
  • For practical, hands-on training, contact your supervisor.