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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

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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.

Autoclave hazards1

Autoclave Hazards

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.

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