Autoclave

From Protists

Contents

Models of Autoclaves

Steris/Amsco SG-120 (New one)

1. Place items to be sterilized into a stainless steel or autoclave-safe plastic tray.

2. Step on pedal at the right bottom corner of the autoclave to open the door.

3. Place trays in autoclave. Loosen any caps/lids.

4. Step on pedal to close the autoclave door.

5. Use the touchscreen to select a program. Check the temp. and time parameters and if appropriate, select "Start Program" and if not appropriate select "Main Menu" to check other programs or select "Change Values" to change the values of the program. However, if you change values, make note of what they were before you change them because you will have to change them back to their previous settings after your run is finished. (Other people who use that program after you and don't check first will be very angry when they realize what happened!)

6. Come back when the program is finished running and step on the pedal to open the autoclave door.

7. Carefully close any liquid-containing objects, then remove tray.

8. Step on the pedal to close the autoclave door so the jacket pressure can be maintained and it will be ready for the next user.


Amsco 3011 (Old one)

1. Place items to be sterilized into a stainless steel or autoclave-safe plastic tray.

2. Place trays in autoclave. Loosen any caps/lids.

3. Close door and turn wheel until hard to turn. The display will indicate that the door is open, but even when the display goes away, continue tightening.

4. Select one of the four options on the display (numbers 1-4). Press number button one time to see what the temp. and times are.

5. Once you know which #, press the # button again (select same # button twice). Otherwise, select the button once and then lift the black cover to reveal more buttons and select "Change Values" and then change them appropriately to match the values of the program you need. However, if you change values, make note of what they were before you change them because you will have to change them back to their previous settings after your run is finished. (Other people who use that program after you and don't check first will be very angry when they realize what happened!)

6. Once inside the chamber pressure is at zero, the diplay will tell you to "Warning, Hot liquids. Open door 1"; Wait 10 min."

7. Carefully close any liquid-containing objects, then remove tray and hit the "RESET" button so you don't have to wait 10 minutes and it is ready to go for the next user.

Types of Runs

Gravity (Dry)

  • 15min sterilizing time
  • no exhaust time should be necessary
  • might have drying time built in though (~10-15min)


Liquid

  • 30min sterilizing time/0.5L (or less) media/container
    • 100mL = 0.1L = 30min
    • 400mL = 0.4L = 30min
    • 500mL = 0.5L = 30min
    • 1000mL = 1.0L = 1hr
    • 1500mL = 1.5L = 1hr30min
    • 2000mL = 2.0L = 2hrs
  • ~15min exhaust time necessary so liquids don't boil over/out
  • no drying time should be necessary


Autoclave Multiuser Courtesy

  • NEVER stop someone else's cycle before it is done!
  • Pick up your items in a timely manner!
  • Be patient and make sure your cycle timer actually starts (this means the jacket has reached appropriate pressure and temperature).
  • If you change values, make note of what they were before you change them because you will have to change them back to their previous settings after your run is finished.
  • Do not leave anything in the autoclave room
  • Dispose of biohazardous waste bags in the trashcans that are provided and do not overfill them (bring them down to our lab trash cans if necessary)
  • Use trays under autoclaving materials in case of breakages, melting, etc. so that the autoclave doesn't get stopped up or broken
  • Don't overfill biohazardous waste bags. You should be able to close the bag. Loosely tape the bag (or use a rubber band). Do not twist the bag, or it won't sterilize properly if the bag is sealed.
  • Dispose of biohazardous waste bags in the trashcans that are provided and do not overfill them (bring them down to our lab trash cans if necessary)
  • See Safety section...


Safety

  • Make sure plastic materials are compatible with being autoclaved.
  • Make sure the autoclave door is fully closed and latched and the correct cycle is selected before starting the cycle.
  • Wear heat resistant gloves when opening the autoclave door after a cycle and while removing hot items.
  • If the door must be opened and the "Hot Liquids" warning is on the display, wear the heat resistant gloves and even eye and face protection. Then, stand behind the door when opening and beware of a rush of steam (only open a little at first to let the majority of the steam escape up instead of out and in your face).
  • NEVER put solvents, volatile or corrosive chemicals (such as phenol, chloroform, bleach, etc.) or radioactive materials in an autoclave.


REMEMBER: When you autoclave, make sure that you leave all caps or coverings slightly unscrewed or loosened so that pressure won't build up inside the tubes or bottles. When finished autoclaving, immediately tighten all caps/covers to prevent contamination. However, DO NOT overtighten or we will not be able to loosen the cap later when it is cooled (if this happens, leave stuck-shut glassware in the transfer hood while working with the flame on for awhile and it should warm up a little to make it easier to loosen). If you forget to loosen your caps, your glassware will explode and break the autoclave. The same goes for anything with aluminum foil...don't tighten it until it has finished autoclaving.


If anything happens while you are working with the autoclave or if you go to use one and notice it is malfunctioning, ask other people in labs by the autoclave or other members of our lab for help. If there is a bad message, then write it down and ask Kathy Vinson in the Cellular Biology Office (room 724) to contact the autoclave repairman and relay the message to him.

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