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    Practical

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    Sana Noor
    The document discusses the five steps in culturing microbes: inoculation, isolation, incubation, inspection, and identification. Innoculation involves introducing bacteria into a growth medium using aseptic technique. Isolation involves streaking colonies on differential media to obtain a pure culture of one microbe. Incubation allows microbes to grow under optimal conditions. Inspection involves observing colony morphology and Gram staining. Identification involves correlating observation data to identify the microbe to the species level. The document also compares light and electron microscopes, noting their differences in energy source, cost, magnification, and viewing methods.

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    Practical

    Uploaded by

    Sana Noor
    4 views25 pages
    The document discusses the five steps in culturing microbes: inoculation, isolation, incubation, inspection, and identification. Innoculation involves introducing bacteria into a growth medium using aseptic technique. Isolation involves streaking colonies on differential media to obtain a pure culture of one microbe. Incubation allows microbes to grow under optimal conditions. Inspection involves observing colony morphology and Gram staining. Identification involves correlating observation data to identify the microbe to the species level. The document also compares light and electron microscopes, noting their differences in energy source, cost, magnification, and viewing methods.

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    The document discusses the five steps in culturing microbes: inoculation, isolation, incubation, inspection, and identification. Innoculation involves introducing bacteria into a growth medium using aseptic technique. Isolation involves streaking colonies on differential media to obtain a pure culture of one microbe. Incubation allows microbes to grow under optimal conditions. Inspection involves observing colony morphology and Gram staining. Identification involves correlating observation data to identify the microbe to the species level. The document also compares light and electron microscopes, noting their differences in energy source, cost, magnification, and viewing methods.

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    Download as ppt, pdf, or txt
    4 views25 pages

    Practical

    Uploaded by

    Sana Noor
    The document discusses the five steps in culturing microbes: inoculation, isolation, incubation, inspection, and identification. Innoculation involves introducing bacteria into a growth medium using aseptic technique. Isolation involves streaking colonies on differential media to obtain a pure culture of one microbe. Incubation allows microbes to grow under optimal conditions. Inspection involves observing colony morphology and Gram staining. Identification involves correlating observation data to identify the microbe to the species level. The document also compares light and electron microscopes, noting their differences in energy source, cost, magnification, and viewing methods.

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    Microbiology

    Culturing Microbes
    The Five “I’s
    Innoculation: Producing a pure culture
    Isolation: Colony on media, one kind of microbe,
    pure culture
    Incubation: growing microbes under proper
    conditions
    Inspection: Observation of characteristics (data)
    Identification: use of data, correaltion, to ID
    organism to exact species
    Microbiology
    Culturing Microbes
    The Five “I’s
    Innoculation: Producing a pure culture
    Introduce bacteria into a growth medium using “aseptic technique” to
    prevent contamination. Tools: Bunsen burner, loop. Needle, etc.
    Microbiology
    Innoculation: Producing a pure culture
    Introduce bacteria into a growth medium using “aseptic technique” to
    prevent contamination. Tools: Bunsen burner, loop. Needle, etc.
    Microbiology
    Isolation: Colony on media, one kind of
    microbe, pure culture: isolation on general
    and special “differential media”
    General growth media: NA, TSA
    Differential: Mac, EMB, SS
    These have dyes, salts, inhibiting
    agents : see differences on
    plates
    Microbiology
    Isolation: Colony on media, one kind of
    microbe, pure culture
    Microbiology
    Isolation: Colony on media, one kind of
    microbe, pure culture – Streak Plates
    Microbiology
    Isolation: Colony on media, one kind of
    microbe, pure culture. Many colonies? Use
    a needle, pick one, and redo streak plate
    Microbiology
    Differential: Mac, EMB, SS
    These have dyes, salts, inhibiting
    agents : see differences on plates
    Microbiology
    • Blood agar : rich with nutrients, can see a
    difference, thus differential; much more
    later
    Microbiology
    • Incubation: Allow organisms to grow under the optimal
    conditions
    • Temperature, with or without oxygen etc
    Microbiology
    • Incubation: Allow organisms to grow under the optimal conditions
    • Temperature, with or without oxygen etc
    • Candle jar reduces oxygen
    Microbiology
    • Inspection: Observation, description
    • Colony Morphology, Microscopic examination (grams
    stain)
    • Systematic recording of “DATA”
    Microbiology
    • Microscopic study: Gram + bacilli, Gram -
    bacilli
    Microbiology
    • Microscopic study: Acid fast, and capsule
    Microbiology
    • Identification: Correlating data from all observations to
    ID organism to species
    • Resources: flow charts, Bergey’s manual etc.
    • Ex. Gram – bacilli, ferments lactose, green sheen on
    EMB: E.coli
    Microbiology
    • Identification: Correlating data from all observations to
    ID organism to species
    • Gram + cocci, grape like clusters, golden yellow colonies, catalase
    +, coagulase +, resistant to Methicillin (MRSA)
    • Staphylococcus aureus
    Microbiology

    Microscopy
    Light microscope: Visible Light is the energy source
    Microbiology

    Light can be described as a form of energy that moves in “waves” .


    Wavelengths of light in the visible spectrum are used in most
    microscopes. Remember the “prism”? Light is composed of different
    colors of light. Each color has different wavelength. Longer
    wavelengths have less energy (red end). Shorter; more energy (violet to
    UV).
    Microbiology
    When light strikes an object the light can be:
    Reflected – Bounces off (Mirror)
    Transmitted – Passes through (GLASS)
    Absorbed – Soaked (black colored paper)
    Diffracted – Scattered as it passes through
    (bugs on a dirty windshield)
    Refracted – Bent as it passes (objects seen
    under water) Glass lenses
    Refractive index: degree of bending,
    based on lens material and shape
    of lens
    Microbiology
    So What? It is a big deal. When light in a scope strikes an
    object (stained bacteria on a slide) some of the light is:
    Absorbed A pattern is collected by the lenses and our
    Refracted eyes see a magnified “object”
    Diffracted
    Reflected
    Transmitted
    Microbiology

    • Types of Light Microscopes


    – Brightfield – most common, objects are dark
    against a bright background
    – Darkfield - special condenser, objects are
    light against a dark background – used to see
    live microbes unstained (spirochetes in fluid)
    – Phase contrast – expensive condenser and
    internal lens components, change “phase of
    light”, so live specimens appear with more
    internal contrast
    – Fluorescence – fluorescent dyes and UV light
    Microbiology
    • Electron Microscope: energy source for magnification is a beam of
    electrons (negative charged subatomic particles
    Microbiology

    • Transmission electron microscope – very


    high magnification (100,000 X)
    • Scanning: tremendous surface detail
    • Transmission Scanning
    Microbiology
    • Compare and contrast Light and Electron Microscope
    • Light Electron
    • Energy – light Energy – electron
    beam
    • Cost - $1200 Cost – $120,000
    • Simple to use Complex processes.
    trained technician
    • Magnification – 1200X Magnification –
    100,000X
    • Viewed by eye, camera Viewed with CRT,
    photos
    Microbiology
    • Compare and contrast Light and Electron Microscope

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