AUTOCLAVE  OPERATING  PROCEDURES

 

IMPROPER  USE  OF  THE  AUTOCLAVE  IS  DANGEROUS.  It produces high temperature, pressure and steam.   Repairs due to improper use are expensive.

 

Autoclaving is a process used to destroy microorganisms and decontaminate infectious biowaste.

 

Training is essential to use the autoclave and minimize the risk of injury. It will ensure that sterility of materials required for research is achieved and that decontamination of all biological materials is achieved. Training will be done by Caroline Golt (Room 112 Worrilow Hall). Training by anyone else (fellow lab mates) is unacceptable.

Wear proper attire when using autoclave – safety glasses, lab coat, closed toe shoes, and insulated gloves for protection.

Power to autoclave should be ON all the time – if panel is not lit – check that POWER switch and control switch on panel are ON. If they do not operate contact Caroline Golt in 112 Worrilow for service. Do not attempt to use autoclave if there is a problem.

If you hear an alarm ringing PRESS RESET button on control panel.

Place all of your items in an autoclavable tub with sides at least 5” high which can contain twice the volume of liquid in case there is boilover or breakage. Dry items should also be placed on trays or tubs.

Make sure that the plastic tub is polypropylene designed for autoclave use.

Strat-Line autoclave tape will indicate when items have been autoclaved.

To autoclave waste – place items in autoclaveable bag and place bag inside a tub making sure that the bag does not contact the hot sides of the autoclave.

Do NOT overfill the autoclave. Overloading hinders steam penetration. The items in the center of the tray may not reach high enough temperature for long enough to kill microbes.

TIME for length of cycle is set by turning the thumb wheel on the front panel to the selected setting. Check to see that the digital display corresponds to the time set.

Do NOT change any other settings on the autoclave!!!

Place items in autoclave carefully to avoid burns. Center items on rack and make sure that there is plenty of room for steam circulation.

Close door making sure that upper left side is pushed in as far as possible.

Tighten door clockwise until DOOR UNLOCKED light is no longer lit. If light does not go out after door is reasonably tight – loosen door and try again – door switch may need repair.

PRESS LIQUID OR GRAVITY (more details below) to begin cycle.

WAIT 1 minute to make sure that no steam is escaping from door area and that pressure is increasing (upper right of control panel).

Write your NAME and ROOM NUMBER legibly on the paper tape for your cycle.

At end of cycle LED panel will indicate CYCLE COMPLETE – wait until pressure reading (upper right of panel) is less than 1 lb.

BEEP will sound at end of cycle. Press RESET button.

The autoclave produces steam which can burn you when the door is opened. Stand behind autoclave door (to the left side) as you open it to use door as a shield. Loosen door slowly counterclockwise just 1 inch and wait for steam to be released. Open door fully to remove tray making sure that your hands and arms are protected from burns.

Use caution when removing liquids. Sudden full opening of the door can cause liquids to boil over or bottles to break. Let them cool slightly in the autoclave so that bumping does not occur. Do not remove anything from the autoclave that is still boiling. Wait until load has cooled.

Remove all items promptly at end of cycle. Do NOT leave items in autoclave overnight. Gently push autoclave door closed before you leave the area. It wastes energy to heat the room with the autoclave while running the building airconditioner.

WASTE should be cooled and placed in disposal boxes in Room 308. Do NOT leave trays of waste in this room.

DO NOT autoclve liquids containing bleach, formalin or glutaraldehyde.

Do NOT autoclave flammable or volatile liquids.

Do NOT autoclave radioactive materials.

Do NOT autoclave items containing corrosives (acids, bases, phenol).

CYCLE CHOICES

Sterilization conditions differ with the type of materials to be sterilized.

LIQUID cycle is for liquid media in autoclavable glass containers. NOTE: only fill container half full and loosen cap to prevent boilovers and breakage. For screw cap containers make the lid hand tight and then loosen the lid one-half turn. For length of cycle follow recommendation of media manufacturer – usually 15 – 20 min. No dry step is used for liquid cycle.

GRAVITY cycle is for all dry items – tips, flasks, tubes, etc. Time for cycle will depend on volume of material. There are extra wire shelves on the side of the autoclave so that several layers of trays may be loaded. A dry time may be added to cycle for dry loads.

WASTE – place items in autoclavable bag. Do NOT seal bag tightly – it will blow up like a balloon and make a mess in the autoclave. Place bag in an autoclavable tray. Autoclave for 60 minutes usually on gravity cycle but it may depend on type of waste. This is a gravity autoclave and it requires a longer time for sterilization than a prevacuum autoclave. Any questions see Caroline Golt.

If you make a mess – you are responsible for cleaning it up. If you need help please see Caroline Golt in Room 112 Worrilow x2627 or Brian Ladman in 315 Worrilow phone x8734.

In case of emergency call Occupational Health & Safety at x8475 or emergency at 911. Make sure that Caroline Golt x2627 is aware of any problems that occur with the autoclave. There is an emergency phone in the hallway near restrooms.

Sterilization: a preventative treatment – so that viable microorganisms are destroyed prior to being used in experimental procedures.

Decontamination: reduction of biological contamination to a level which no longer poses a risk to persons or the environment.