How to Use an Autoclave

Would you want a surgeon to use a
scalpel that has bacteria growing on it? Of course not! Surgical instruments must be sterilized
that means no living organisms are present. There are lots of products that need to
be sterilized. Pharmaceuticals… equipment… solutions, medical waste, even
tattoo needles. If bacteria, fungi, or viruses are
present the result could be deadly. So how do we sterilize materials in the
lab? We use a device called an autoclave. Basically an autoclave is a powerful
pressure cooker that uses heat, steam, pressure to destroy microorganisms. First we’ll sterilize as a solution using the
slow exhaust cycle And second, we’ll sterilize some empty bottles using
fast exhaust. We need to sterilize this flask
containing an agar bacterial media. Cover the top with foil and place a
piece of autoclave tape over the foil. Autoclave tape looks like regular
masking tape the difference is that it has a special
ink that causes black diagonal lines to appear when exposed to high temperature. Turn on the power and make sure the
drain valve is closed. Add de-ionized water to the level
indicator line. Place the flask of culture media into
the basket. Insert the basket, close the lid, and turn the handle to create an
airtight seal. Use the control panel to set the mode to
sterilize, the temperature to 121 degrees celsius and the
pressure to 15 psi. The slow exhaust mode prevents the
liquid from boiling over and out of the flask. Run the autoclave for
at least 15 minutes. While it’s running, enter the date and time
and the operation details in the log and initial it. Once the cycle ends and the
pressure gauge reads 0 psi slowly open the door wearing heat
resistant gloves. The black lines are visible on the
autoclave tape indicating that the proper temperature was reached. Once the culture media has cooled to
about 45 degrees celsius it’s ready to be poured into petri dishes. Now let’s run the second autoclave cycle
this time using the fast exhaust to sterilize some empty bottles. Tighten the cap, but then loosen
it about half way. Place the autoclave tape over the lid
and put the bottles into the basket. Set the autoclave to 121 degrees celsius,
15 psi… and a 20-minute runtime. Select the fast exhaust mode. Complete and initial the log. Once the cycle ends and the pressure gauge
reads 0 psi, slowly open the door and remove the basket the black lines are visible on the tape
indicating that the proper temperature was reached. The autoclave is now ready for another
run. Remember slow exhaust is for liquids
and medical waste. Fast exhaust is presented glassware an
instruments. Now you’re ready to go to the lab and
use an autoclave.

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23 Responses

  1. Jesus Palacios says:

    Thank you for the video. Do you know any FDA or ISO guideline for ussing propperly the autoclave? Thanks.

  2. Ammar Hussain says:

    Thank you very much… it is really a good video to understand the basic mechanism of autoclave… GOD bless you
    muffin choc
    answer to your question is "yes",
    you would use the same procedure but "fast exhaust" one because you are just sterilizing beakers (which are empty glasswares)…
    remember… liquid turns solid slowly, so slow exhaust for liquid media's
    and …. solid melts fast, so fast exhaust for solid things (i.e. instruments or empty glass wares)

    Dr. Ammar

  3. Jalal Alakhras says:

    thanks very much ,, it helped me a lot 

  4. Nudrat Fatima says:

    very easy for us to explain students

  5. Bill K says:

    Important Videos For New Tattoo Artist!
    Take Care, Bill Kearney 🙂

  6. Gediminas says:

    Is vacuum also used?

  7. UsedStryker says:

    An autoclave is an expensive medical tool and should be handled properly. If you are in doubt on what to do, always consult the user’s guide.

  8. Elouise Gaylard says:

    is it 121F ? struggling here in Aus

  9. Tomson says:

    love educational videos like this helped me alot:)

  10. akhan shamenov says:

    Using autoclave is important when preparing for a surgical procedures. I wonder if it uses more energy than kettle or microwave.

  11. angela says:

    when should I turn off the autoclave? after or before opening the lid once the cycle is done? help please:)

  12. vveenaaa says:

    the embed code doesnt works in powerpoint .pls help

  13. Jillian says:

    Cramming for a certification exam tomorrow. Thank you!

  14. Frederick-Nrunk Kamara says:

    Autoclaves have come along way since I autoclaved media broth

  15. Diarmuid Mac Mahon says:

    how does the pressure clean the inside of the containers? surely some extremophiles type bacteria could live on the inside of the sealed containers

  16. Goku says:

    These still can't destroy prions! D:

  17. A jay Johnson says:

    Fun Gee

  18. Binny Kakkassery says:

    Is this use for retort pack

  19. Autoclave Depot says:

    Nice job mate!

  20. Raisha Haddad says:

    I love and supported the people who making this educational videos ???

  21. Cebidex Pharmaceutical says:

    thanks fast and to the point information…

  22. Panda ReefNguyen1994 says:


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