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Aseptic Technique: Media and Equipment

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Presentation on theme: "Aseptic Technique: Media and Equipment"— Presentation transcript:

1 Aseptic Technique: Media and Equipment

2 Growth Medium A growth medium or culture medium is a liquid or gel designed to _______ ____ _______ of microorganisms or cells The most common growth media for microorganisms are ________ _______ and _______ plates

3 Defined Media An important distinction between growth media types is that of ________ versus __________ media A defined medium will have _________ quantities of all ingredients

4 Undefined Media An undefined medium has some complex ingredients that consist of a mixture of many chemical species in _________ _____________ Undefined media are sometimes chosen based on _______ and sometimes by ____________ - some microorganisms have never been cultured on defined media.

5 Undefined Media We will use this in the lab Nutrient Agar contains:
________ _______ ___ ________ ________ ________ ________

6 Forms of Culture The most common growth media for microorganisms are ________ _______(liquid in a test tube) Liquid media are often mixed with ______ and poured into ______ _______ to solidify These agar plates provide a _______ medium on which microbes may be cultured. They remain solid, as very few bacteria are able to decompose agar.



9 Slants There are several reasons a slant is best for culture storage:
1) ________ ______ – slant increases it 2) ______ – test tubes smaller to store 3) ________ – won’t dry out 4) _______ – once the microorganism is transferred, you can store it for up to 6 months in a refrigerator


11 Aseptic Technique Aseptic technique refers to a procedure that is performed under ________ ____________ This includes ________ and _________ techniques (such as with microbiological cultures) It includes techniques like _________ ______________ (Bunsen burner)

12 Aseptic Technique Sterile surfaces must be protected from ________ ___ ____ ____ or on non-sterile surfaces In sterile technique, only ________ surfaces touch other ________ surfaces and ____ exposure is kept to a minimum

13 Aseptic Technique It is important in microbiology to work with ________ __________ This is difficult because the world around us is covered with ________ (even on dust particles in the air) In order to protect broth, plates, slants and pure cultures from MO’s around us, we practice _________ __________

14 Aseptic Technique In this class you will need to practice sterile technique when we _________ a pure culture into ______ __________ (tube of sterile broth or an agar plate)

15 Aseptic Technique “A” = Negative prefix “Septic” = Infection
All techniques and procedures which __________ _______________

16 Safe Patterns Safe patterns include: Standard _______ _________
______________ Techniques ______ and ________ of Equipment _________ Hands

17 Safe Patterns ________ all materials before beginning

18 Safe Patterns Spray the lab top down with a _________ _________ or a ______ _______ __________ and allow this to stand for a _______. You may then wipe down the bench with the paper towel.

19 Safe Patterns Wash hands _______ and _______ lab.

20 Safe Patterns You should have only the _________ ______ ________ and the written _____ __________ on your bench top or desk.

21 Safe Patterns Keep petri dishes and test tubes ___________ as much as possible. If top must be removed completely do ______ _____ _____on the lab top. This lowers the probability of contamination and prevents “false positive” results.

22 Safe Patterns Hold bottles and tubes ____ ___ _______ to minimize the amount of airborne microbes that can fall into them (blue circle). Remove the caps as shown above and ___ _____set the caps down. Keep the mouth of the cap facing _________ (red circle).

23 Safe Patterns When using metal ________ ________, HEAT the _______ piece of metal of the inoculation instrument in the flame: it should be RED HOT. Be sure to ________ your inoculation instrument ________ picking the inoculum (broth or agar).

24 Safe Patterns To __________ a Petri plate: Lift one edge of the Petri plate cover to gain access to the culture medium. Keep the cover over the plate bottom to prevent ______ and __________ from falling onto the agar.

25 Safe Patterns Report _________ to me immediately
Cover the spill with ______ ________ and squirt _________ onto the towels. Wait _____ _________ then clean up the spill.

26 Safe Patterns ________ all test tubes and petri plates with your name (initials), date, and name of organism ________ you add any solutions, bacteria, etc. 

27 Safe Patterns Do _______ dump ANY microbial suspension down the ______ or in the _______ ______. I will collect them for proper disposal.

28 Safe Patterns Place _____ ________in racks when working at your table:
never lay the tubes down—____ _____ Keep test tube ______ and petri dish _______ on media to reduce _______________ (matters not whether it is sterile media or already cultured).

29 Safe Patterns All agar plates are incubated _______ ______
to reduce bacterial _______________ and to reduce the possibility of water _______________ that may be on the lid dropping onto the agar, causing fluid to run across the agar medium.  

30 Media __________ after preparation, in storage, or working containers. Handle _____________. If no goofs – Sterile ___________

31 E. Autoclave _____________ ________
Water boils at 100ºC, but if pressure is __________ the boiling point _______ At 15 PSI the Boiling Point = 121ºC At 20 PSI the Boiling Point = 126ºC Minimum = ____ ____ PSI A pressure cooker is the same as an ____________

32 Boiling _________ kills _______ MO, fungi, and viruses but a few are ____________. Example: Inf. Hepatitis, Endospores But very good nonprofessional use

33 Technique depends on article:
Equipment Technique depends on article: Does it _____ or _____? Do you want it back?

34 Equipment Autoclave If it _____ into machine and ____ ____
___ 121ºC (15 PSI) minimum

35 Equipment Boiling ________ in a lab

36 Equipment Oven _____ ______ used for metal and glass
____ _________ minimum

37 Equipment ___________ – Burning ________ Plus Loops, test tubes

38 Equipment _____________ Equipment Usually _______ and ______ materials
Gas: _______ _______ ____________

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