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Introduction to the Microbiology Lab

Introduction to the Microbiology Lab. Exercises 1-2, 2-1, 3-1, and 3-3. Laboratory Safety. Bio Safety Levels. Student Conduct. Do NOT smoke, eat, drink, or bring food or drinks into the laboratory room Do NOT apply cosmetics or handle contact lenses

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Introduction to the Microbiology Lab

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  1. Introduction to theMicrobiology Lab Exercises 1-2, 2-1, 3-1, and 3-3

  2. Laboratory Safety

  3. Bio Safety Levels

  4. Student Conduct • Do NOT smoke, eat, drink, or bring food or drinks into the laboratory room • Do NOT apply cosmetics or handle contact lenses • Wash your hands thoroughly with soap and water after handling microbes AND before leaving the laboratory room each day • Do NOT remove organisms or chemicals from the laboratory • Work carefully and methodically • COME PREPARED TO LAB FOR THAT DAY’S WORK

  5. Basic Laboratory Safety • Wear your lab coat and gloves while working in the laboratory • Remove them before leaving • Discard gloves appropriately (not in standard trash) • NO OPEN-TOED SHOES OR SANDALS • Turn off Bunsen burners when not in use • Secure loose hair and clothing when working with an open flame • Never pipette by mouth

  6. Safety Equipment • Know the location of the First Aid Kit • Know the location and proper way to use the Fire Blanket, Shower, and Fire Extinguisher • Know the location and proper way to use the Eye Wash Basin • NOTIFY YOUR INSTRUCTOR OF ANY EVENT THAT NECESSITATES THE USE OF THESE

  7. Reduce Risk of Contamination • Clean bench with disinfectant before and after lab • 70% alcohol can also be used as a disinfectant • Allow to air dry rather than wipe up • Never lay culture tubes on bench top • To clean spills: cover with paper towels, saturate with disinfectant and leave for 20 minutes • Discard in appropriate container • Place non-essential books, etc. out of the way of your experiments

  8. Disposing of Contaminated Materials • Broken glassware, needles, wooden cotton swabs, etc. MUST be placed in the “sharps” container • Culture tubes are to have any labels removed and placed upright in racks on the “kill” cart • Petri dishes can be discarded into solid waste on the “kill” cart WITHOUT removing the labels • Gloves are to be discarded in the appropriate biohazard containers

  9. Care of Microscopes • Microscopes are located at each lab station • Following their use, microscopes: • Should have all oil removed from the lenses using LENS PAPER and cleaning solution • YOUR INSTRUCTOR WILL INSPECT THEM BEFORE YOU ARE PERMITTED TO LEAVE • Scanning (4X) lens should be in place • Stage should be at lowest setting • Rheostat should be adjusted to its lowest setting • Cord should be wrapped carefully around the base and not “flopping” around

  10. Care of the Lab • Place seats back under benches before you leave • Wipe down the sink-tops with paper towel if you are among the last students in the lab • Remove any paper towels left behind by less conscientious students THE LAB SHOULD LOOK AS NICE AFTER YOU ARE DONE AS IT DID BEFORE YOU STARTED

  11. Scavenger Hunt

  12. Essential Equipment • Security Phone • Disinfectant • Antiseptic Hand Solution • Fire Extinguisher • Emergency Shower • With Eye Wash • Fire Blanket • MSDS Folder • Biohazard Waste Container • Biohazard Sharps Container • Broken Glass Container • “Kill” Cart • Incubators • Refrigerated Cold Room • Biological Safety Cabinet • Vortex Mixers • Microscopes • First Aid Kit • Chemical Spill Kit • Microbiology Lab Tools • Autoclave • Fire Alarm • Escape Routes • Gas Shut-off

  13. New Experiments

  14. GLO-Germ Hand Wash Sanitation System Exercise 1-2

  15. Glo Germ Hand Wash System • Effective hand washing minimizes transmission of pathogens by health-care workers and food handlers • How Effective Is Your Hand Washing Technique? • Follow the directions outlined in the lab manual… • Work in groups of 2 with one person applying the Glo Germ gel and the other person holding the UV lamp • Do NOT hold UV lamp if you also have Glo Germ applied • TURN UV LAMP OFF WHEN EXERCISE IS COMPLETE

  16. The Ubiquity of Microorganisms Exercise 2-1

  17. Ubiquity of Microorganisms • Many microbes can easily be isolated from a variety of environments • Soil, water, plants, and animals • Just how ubiquitous are microbes? • Follow the directions outlined in the lab manual… • You will work in groups of 4 • Incubate plates 1, 2, and 8 at 25oC. • Incubate plates 3, 4, 5, 6, and 7 at 37oC.

  18. Introduction to the Light Microscope Exercise 3-1

  19. Principles of Light Microscopy • Illumination • Stains add contrast to specimens • Magnification • Compound microscope has paired ocular lenses and an objective lens • Resolution (Resolving Power) • Clarity of an object viewed under the microscope • Distance that two points must be apart to appear as two distinct points • Resolution of typical light microscope is 0.2μm

  20. Parts of the Light Microscope Involved in Illumination Involved in Magnification/Focus Stage Stage Mechanisms Course/Fine Adjustment Revolving Nosepiece Objective Lenses 4X, 10X, 40X 100X (Oil Immersion) Ocular Lenses • Base • On/Off Switch • Lamp (Bulb) • Light Intensity Knob • Sub-stage Condenser • Iris Diaphragm

  21. Microscopic Examination of Eukaryotic Microbes Exercise 3-3

  22. Examination of Eukaryotic Microbes • Follow the directions outlined in the lab manual… • You will work individually with your own microscope, but feel free to divide the specimens up equally for purposes of seeing everything: Oscillatoria sp. Amoeba Saccharomyces cerevisiae Giardia lamblia Aspergillussp. Spyrogyra sp. Rhizopussp. Diatoms Euglena sp. Volvoxsp. Paramecium caudatumDog tapeworm Human louse Plasmodium sp. (in RBCs)

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