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How a laboratory autoclave works

An autoclave is a pressure vessel that sterilizes with saturated steam. Steam under pressure is hotter than boiling water at normal pressure, and it carries heat to surfaces far more effectively than dry air.

The basic conditions

A common cycle holds the load at about 121 degrees Celsius (250 degrees Fahrenheit), at roughly 15 pounds per square inch of pressure above atmospheric, for at least 15 to 30 minutes. The time depends on what you are sterilizing, how large and dense the load is and your institution's validated procedures. Always follow the manufacturer's instructions and your own protocols.

Stages of a cycle

  1. Air removal. Air must leave the chamber, because trapped air creates cold spots.
  2. Heat-up. Steam raises the chamber and load to temperature.
  3. Sterilization hold. The target temperature is held for the set time, which starts once the load itself reaches temperature.
  4. Exhaust. Steam is released. Liquids need a slow exhaust so they do not boil over.
  5. Drying, if the cycle includes it. Used for wrapped instruments.

Types of autoclave

TypeHow it removes airTypical use
Gravity displacementSteam pushes air out of a drainMedia, glassware, waste, many general loads
Pre-vacuum (high vacuum)A pump removes air before steam entersWrapped instruments and porous loads
Benchtop vs floor-standingSame principles, different sizeSmall labs vs larger throughput

Why loading matters

Steam has to reach every surface. Overpacked loads, sealed containers and trapped air can leave the inside of a load below sterilizing temperature even when the chamber gauge reads correctly.

Check yourself with the pre-run checklist.
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