Autoclave Instructions

Autoclave Instructions 

Autoclaves use high temperatures and pressure to inactivate biologically active material to ensure it is non-viable prior to waste disposal. See below for directions on how to safely operate the autoclave.  

Before use 

  1. Ensure no items have been left by previous users, especially sharps or other items that could pose a hazard. Remove these items or contact NSM or the Biology technical staff to have items removed.

  2. Clean the strainer before loading the autoclave. 

  3. Ensure the material to be autoclaved does not contain any hazardous or radioactive material. Contact EHS at x7233 for instructions on how to dispose of hazardous or radioactive material. 

  4. Loosen bottle caps before loading to prevent them from shattering; DO NOT over-fill liquids; ensure containers are free of cracks. 

  5. Ensure any plastic materials are compatible with the autoclave, so that they maintain their integrity with autoclave temperatures and pressure. 

  6. Place solids into a labeled "autoclave bag"; ensure that solids have at least 100 ml of liquid present with the load to effectively sterilize them. 

  7. Place a metal drip pan underneath items to catch spills. 

  8. Place autoclave indicator tape on every item autoclaved and check the tape to see if a color change has occurred after the run is complete. The indicator tape confirms that the run achieved the proper temperature and pressure to inactivate biological material. 

  9. Close the autoclave door before using. 

During use 

  1. Autoclave when there are fewer people around to minimize odor complaints.

  2. Set the proper time for the material that is being autoclaved. Most liquids and dry goods can be autoclaved in 15 minutes. Check the protocol.

  3. Autoclave waste liquids or solids for 45 minutes to properly inactivate biological material.

After use 

  1. Use thermal protective gloves, apron, and face shield to protect against heat.

  2. Handle carefully: liquids can "bump" or suddenly erupt and spill when the container is moved.

  3. Use the drip pan when removing the bags to avoid spills.

  4. Clean up all spilled liquid inside or outside the autoclave. If over a liter, obtain a spill kit from one of the labs. Spill kits contain absorbent material that will assist in the cleanup.

  5. Clean the filters/traps using tap water. Traps are located on the floor of the autoclave.

Dispose of autoclaved waste properly 

  1. Liquids can be disposed down the drain as long as no hazardous materials are present.

  2. Do not drain dispose agar or any material that can solidify and clog the drain. Solidify agar in a beaker and put in the regular trash.

  3. Place the autoclave bags into regular opaque plastic bags and take directly to the outside dumpsters.

Specific instructions to turn on the autoclaves in DBH 

  1. Press the Controls 'ON' switch. The green indicator in the upper left corner of the switch will light.

  2. Open the door. The Unsealed Door Indicator will remain lit until the door is closed and sealed. If the Unsealed Door Indicator is not lit, the door will not open, press Unseal Door Switch.

  3. Load the autoclave, and then close the door.

  4. Select Processing Cycle. Wrapped (looks like a burrito), Unwrapped (scissors) or Liquids (flask).

  5. Select Exposure Temperature (in C). Usually this is 121 ⁰ C. Note: Check the temperature. Never autoclave things over 130⁰ C.

  6. Select Exposure Time Hours are displayed at the left of the color; minutes at the right. For example, 02:30 means two hours and thirty minutes. The time selection will remain displayed until it is changed.

  7. Select Exhaust (Drying) If no drying time is desired, set the Exhaust (Drying) Time Display to 00:00. If you are autoclaving liquids, you cannot change the Exhaust time. It is preset.

  8. Use care in opening the door. Open slightly to vent steam then open fully to remove items while wearing PPE.