Autoclave Operation, Functioning and Cleaning

Laboratory autoclave – loading​

Before Autoclaving

  • Before autoclaving, make sure that the items you are autoclaving are safe to autoclave. Anything that emits toxic gases should not be autoclaved. For example, do not autoclave bleach. Check under “Safety” subheading above for items you can’t autoclave.
  • Inspect glassware for cracks. Do not autoclave cracked glassware. Glassware must be made of borosilicate.
  • Prepare and package autoclavable items properly. Loose dry materials should be wrapped or bagged in appropriate steam-penetrable paper. Do not wrap tightly as this will prevent steam penetration.
  • Make sure all lids and bags are loosened to prevent pressure build up and possible explosion. Containers that don’t have tops must be covered with steam penetrating enclosures such as cotton plugs or steam-penetrable bungs. If containers contain liquids, do not fill them more that 2/3 full.
  • Plastics must be heat-resistant. For example, polycarbonate, Teflon, and most polypropylene plastics are autoclaveable.
  • All items to be autoclaved must have autoclave tape on them.
  • Secondary containers (i.e. stainless steel pans) should be used when containers carry liquids that could spill. This helps to contain spills as well as make it easier to carry multiple items to and from the autoclave. A secondary container must be big enough to handle any spill. Water can be added to the secondary container to prevent heat shock.

Loading an Autoclave

  • You should have your lab coat on, eye protection, gloves, and closed-toe shoes.
  • Place autoclavable materials in an orderly fashion well spaced from each other for proper steam penetration. Make sure all items are compatible. Do not overload.
  • Close the autoclave and make sure the door is firmly latched.

Operating an Autoclave

  • Once the door is locked, choose the appropriate cycle for the material you’re autoclaving. If necessary check the manual that came with your autoclave. All autoclave manuals should be close at hand.
  • Automatic cycles are preset. Just push the appropriate button. Customized cycles require programming. Consult with your supervisor on how to do this if needed.
  • If required to fill out a user log, do so at this time with your contact information.
  • Do not open the autoclave when it is in operation.
  • If the autoclave is not working properly contact your supervisor immediately.

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Unloading an Autoclave

  • Wear protective clothing and gear to prevent burns.
  • Make sure the cycle has completed and that pressure and temperature have returned to a safe level.
  • Carefully open the door and open the door slowly about one inch or so. This releases residual steam while allowing pressure in liquids to normalize to 1 atmosphere.
  • Check autoclave tape for color change. If no color change, that means autoclave cycle failed. The load will need to be re-autoclaved in a device that is working properly.
  • Bags that carry the biohazard symbol need to be over bagged with an opaque trash bag and sealed prior to disposal in regular trash

Autoclave Malfunctions

  • If the autoclave malfunctions, don’t try to fix the problem. The issue needs to be properly diagnosed and corrected by qualified professionals who are authorized to do so.
  • Record the malfunction in the log book.
  • All accidents need to be reported to the supervisor. For example, spills or releases of biohazardous materials or recombinant DNA.
  • Personal injury should be reported immediately. If necessary seek medical assistance. If clothing has been soaked with hot water/steam, remove your clothing and place the injury in cool water.

Spill Clean-up

  • Spills can occur at any time from liquids that have boiled over or containers that have broken during autoclaving.
  • All spills need to be cleaned up before using the autoclave again.
  • The current operator is responsible for clean-up.
  • If you have a spill kit use it, otherwise use paper towels. Make sure the autoclave and materials that have spilled are at room temperature before removal.
  • If uncertain how to clean up the spill, consult you biohazard manual for proper disposal. If several materials have mixed, make sure to use the protocol for the most hazardous component of the mixture.
  • Make sure to dispose of any cracked glassware properly.
  • If required, record information about the spill and the clean-up procedure you used in your log book.