Welcome to Microbiology Lab Tue 6:00-9:10pm Instructor: Huma Malik Office location: MTSC 339 Office hours: Tue 5:00-6:00pm (or by appt.)

Introduction Lab safety rules! Attendance: Don’t be late to class! Make-up lab MAY be possible if they are arranged sufficiently ahead of time and with instructor permission Cheating and plagiarism will not be tolerated! Copying someone else’s work or cheating on quizzes or a practical will result in an “F” grade Use of cell phones is NOT allowed!

Materials and supplies Provided to student: Microscope (assigned according to student’s station number) Numbered slide box with slides Inoculation loop, needle, slide holder, blotting paper, & lens paper Each student must provide the following: Safety glasses Lab coat Gloves  preferably non-latex (nitriles, etc) Masking tape for labeling Colored pencils for recording results Sharpie Lab manual – MUST have by Lab #2!

Grading policies You will receive one grade for Microbiology lecture and lab: Lab = 30% Lecture = 70% 1 Homework assignment10 pts 4 Metrics quizzes (5 pts each)20 pts 2 Quizzes (10 pts each)20 pts 6 (3x5) Cards5 pts Practical I75 pts Practical II (Final Exam)85 pts Water analysis paper35 pts Unknown determination project100 pts TOTAL350

WkDateLab Assignment 118-Feb Laboratory Protocol & Safety; 1 Metric System, 2 Microscopy, 3 Ubiquity of Microbes 225-Feb Read 3 Colony morphology, 4 (A&B) Transfer Techniques (23), 6 Gram stain (HW due) 34-Mar 7 Endospore stain, Read 8B motility tall, 4 (23) Oxygen Req's, 16 Catalase Test (Quiz #1 - metrics) 411-Mar 8A Motility Hanging drop, 9 Acid Fast stain, 4D Streak plates (Quiz #2 - metrics) 518-Mar Read 4D; Demo 10 Capsule; Review Staining Techniques & Colony Morphology (Quiz #3 - metrics) 625-Mar Practicum I 71-Apr Discuss Metabolism & Media; Read Demo 13, Inoculate Exp. 15, 17, 19, 20; & Begin Apr Quiz #4 Lab Media; Read Tests 15, 17, 19, 20 and Demos 18, 21 & 22; Do 24; Cont Apr SPRING BREAK 922-Apr Read 24 and Demos 11, 12, 14, 25, 26, 27; Cont Apr Read and discuss Demos May Quiz #5 Biochemicals, Review - ALL exercises to date! 1213-May Practicum II / BREAK / Orientation, Begin Final Project May Disease Case Studies; Continue 36 Orientation and Progress (Quiz # 6 - metrics) (Water analysis paper due) 1427-May Disease Case Studies; Continue Jun Disease Case Studies; 36 Final Project Due - you should be checked out by June 5 16 Finals - Microbiology Lecture Final is

Lab #1 1. Metric System, 2. Microscopy, and 3. Ubiquity of Microbes Spring 2014

1. Metric System Base units: meter, liter, gram Helps simplify very large and very small values Use dimensional analysis

Dimensional analysis Three steps: Set up the units Put in values Plug and chug Can be used to convert within metric system Can be used to convert between metric system and English units (pound, miles, gallons) Final answers must be in scientific notation

2. Microscopy

Different types of microscopes: Brightfield – we will use these Darkfield Phase contrast Fluorescence Electron microscope Scanning electron microscope Magnification: how large an image looks Resolution: mathematical expression of the ability of a lens system to distinguish detail clearly 2. Microscopy

How to use a microscope? Video: Video on oil immersion: Pg. 10 and 11 of lab manual Key points Start on lowest power (4 or 10X) Use coarse adjustment to bring image into focus Once focused on lowest power then change over to the next objective (40X) – DO NOT MOVE STAGE UP OR DOWN – JUST USE FINE FOCUS Once focused at 40X, put one drop of immersion oil and switch over to the 100X – DO NOT MOVE STAGE UP OR DOWN – JUST USE FINE FOCUS Immersion oil is ONLY used with 100X objective! 2. Microscopy

Properly putting away the microscope: Light off Wrap the cord Stage all the way down On lowest objective (4X) Iris diaphragm open Clean any oil on 100X using LENS paper. Also, clean other objectives with lens paper Clean ocular lenses! DO NOT ROTATE THE HEAD WITH THE OCULAR LENSES! 2. Microscopy

How can you determine the Total Magnification (TM) of a specimen being observed under the microscope? Ocular Lens Objective Lens Total Magnification 10X4X40X 10X 100X 10X40X400X 10X 100X (oil immersion) 1,000X 2. Microscopy

Bacterial cells have certain shapes and arrangements

Bacillus, chains Bacillus, singles 2. Microscopy

Cocci, clusters Cocci, chains 2. Microscopy

Cocci, tetrads 2. Microscopy

Prokaryotes: no nucleus (bacteria and archaea) Eukaryotes: nucleus and membrane bound organelles Protozoa: Giardia, Amoeba, Trypanosoma Unicellular Fungi: Pencillium Unicellular algae: Chlorella Viruses – can’t see with a light microscope AmoebaGiardiaTrypanosoma 2. Microscopy

Each student Find your microscope (same number as your seat in class) Practice using the microscope with prepared bacterial slides: Bacillus/ rod shaped bacteria Cocci/ sphere shaped bacteria Spirilla shpated bacteria Observe eukaryotic cells setup at Demo stations A through D Record shape, arrangement, and TM for each on Pg. 14 and Microscopy

Where are microorganisms found? Each student (Pg. 17): Obtain 1 petri plate with tryptic soy agar (food for microbes) Keep the plate closed with the lid on until ready to use! Take petri plate home (keep closed!) Expose plate to air (30min) or touch an object of interest (fingers, lips, coin, food, etc.) onto the petri plate Put the lid back on Label with what you sampled Place the petri plate in a zip-lock bag for 1 week  bring it to class next week 3. Ubiquity of Microbes