1. Autoclave Use and Safety
University of Massachusetts
Boston
Biology Department
M. Kelly

2. An autoclave is used to sterilize utensils, containers, media, soil, used infectious materials and other waste. Never autoclave toxic, flammable or radiological agents.

3. Standard Temperature and Pressure
121° C (250° F) 15 psi Temperature and pressure are preset – don’t change these Time: varies You can set the autoclave to run for the time you choose.

4. Proper Autoclaving Containers
Containers used for autoclaving must be able to withstand high temperature and pressure without melting or disintegrating.
Use: stainless steel, polypropylene or polycarbonate
If you are unsure whether a container is autoclavable or not, DON’T USE IT
Over time, the integrity of the container decreases

5. Autoclaving Liquids
Use a container that is twice the volume of the liquid you are autoclaving, for example, use a 2 liter flask to autoclave 1 liter of media
Always Always Always use a secondary container under your container of liquid
Loosely cover a container of liquid to be autoclaved with aluminum foil or a cap; never tighten a cover.

6. There are 3 types of cycles (I only use 2): FAST LIQUID DRY
Autoclave Cycles
There are 3 types of cycles (I only use 2): FAST LIQUID DRY

7. Autoclave Cycles
Fast – Fast Exhaust – after autoclaving, valves open and chamber rapidly returns to atmospheric pressure
Use the fast exhaust to sterilize empty glassware, toothpicks, soil, pipettes and metal utensils (anything that does not contain liquid).

8. Autoclave Cycles
Liquid – Slow Exhaust – after autoclaving, valves partially open to allow super-heated liquids to cool slowly and reduce boil-over.
Use the slow exhaust when sterilizing liquids such as media and water. Also use slow exhaust when sterilizing waste plates of solidified media because it will melt in the autoclave.

9. How Long to Autoclave Dry goods @ 121 °C Time (min)
Glassware, empty, inverted
Instruments, wrapped
Utensils, wrapped

10. How Long to Autoclave
Biohazardous waste 121 °C, loosely tied Time (min) 2 or more bags bag, full 90 1 bag, half full 60

11. How Long to Autoclave
Liquids, in bottles with vented caps Size (ml) Time (minutes)

12. How Long to Autoclave The time you need to autoclave will depend upon:
The volume in the flask
The number of flasks in the autoclave
How far apart the flasks are
How high the sides are on the container
The material of the container

13. Loading the Autoclave
Never overload the autoclave; steam needs to circulate
Never autoclave clean items and waste together
Use indicator tape to confirm temperature
Fill in log

14. Unloading the Autoclave
Turn autoclave OFF
Pressure gauge must be at 0 psi to open door
Open door very slowly and stand back from the door
Allow all steam to escape from autoclave
Open door; let materials cool; don’t remove any liquids that are boiling
Use gloves, have feet covered, lab coat

15. Smell
Autoclaved waste usually doesn’t smell good. Be sure to let any waste cool completely in the autoclave room before it is put outside in the trash. Our neighbors in the offices across the hall don’t want to smell trash all afternoon and I wouldn’t either.

16. Cleaning the Filter
Occasionally clean out the filter before you autoclave
Filter is located under the rack that sits on the bottom of the autoclave
Unscrew the nut (when it’s cool) and lift out the small filter, dump out any material in it and replace the nut

17. Troubleshooting
If you have any problems with the autoclave, record it in the log
Tell you supervisor
Leave a note by the autoclave stating what the problem is
If you supervisor is not around you can tell:
Maureen Kelly
Charlie King
Maria Mahoney

18. The Autoclave Room
Please help to keep the autoclave room clean and neat by cleaning up after yourself. Rinse out any containers used for autoclaving. Clean up any spills; put broken glass in the box and sweep up the floor. If you are waiting for your items to finish autoclaving, take a few minutes to clean up the sink or the counter, sweep the floor or put away the racks.

19. Finally
If you are unsure of something we want you to ASK! We appreciate the work you do for us and the department We want you to be safe.