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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.