2 Growth MediumA growth medium or culture medium is a liquid or gel designed to _______ ____ _______ of microorganisms or cellsThe most common growth media for microorganisms are ________ _______ and _______ plates
3 Defined MediaAn important distinction between growth media types is that of ________ versus __________ mediaA defined medium will have _________ quantities of all ingredients
4 Undefined MediaAn 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 CultureThe most common growth media for microorganisms are ________ _______(liquid in a test tube)Liquid media are often mixed with ______ and poured into ______ _______ to solidifyThese 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 it2) ______ – test tubes smaller to store3) ________ – won’t dry out4) _______ – once the microorganism is transferred, you can store it for up to 6 months in a refrigerator
11 Aseptic TechniqueAseptic 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 TechniqueSterile surfaces must be protected from ________ ___ ____ ____ or on non-sterile surfacesIn sterile technique, only ________ surfaces touch other ________ surfaces and ____ exposure is kept to a minimum
13 Aseptic TechniqueIt 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 TechniqueIn 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 PatternsSpray 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 PatternsWash hands _______ and _______ lab.
20 Safe PatternsYou should have only the _________ ______ ________ and the written _____ __________ on your bench top or desk.
21 Safe PatternsKeep 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 PatternsHold 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 PatternsWhen 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 PatternsTo __________ 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 PatternsDo _______ 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 toreduce the possibility of water_______________ that may be on the liddropping onto the agar, causing fluid torun 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 = 121ºCAt 20 PSI the Boiling Point = 126ºCMinimum = ____ ____ PSIA pressure cooker is the same as an ____________
32 Boiling_________ kills _______ MO, fungi, and viruses but a few are ____________.Example: Inf. Hepatitis, EndosporesBut very good nonprofessional use
33 Technique depends on article: EquipmentTechnique depends on article:Does it _____ or _____?Do you want it back?
34 Equipment Autoclave If it _____ into machine and ____ ____ ___ 121ºC (15 PSI) minimum