Welcome to Microbiology Lab

Tue 6:00-9:10pm

Instructor: Huma MalikEmail: Huma.Malik@rccd.eduOffice location: MTSC 339Office 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 assignment 10 pts

4 Metrics quizzes (5 pts each) 20 pts

2 Quizzes (10 pts each) 20 pts

6 (3x5) Cards 5 pts

Practical I 75 pts

Practical II (Final Exam) 85 pts

Water analysis paper 35 pts

Unknown determination project 100 pts

TOTAL 350

Wk Date Lab Assignment

1 18-Feb Laboratory Protocol & Safety; 1 Metric System, 2 Microscopy, 3 Ubiquity of Microbes

2 25-Feb Read 3 Colony morphology, 4 (A&B) Transfer Techniques (23), 6 Gram stain (HW due)

3 4-Mar7 Endospore stain, Read 8B motility tall, 4 (23) Oxygen Req's, 16 Catalase Test (Quiz #1 - metrics)

4 11-Mar8A Motility Hanging drop, 9 Acid Fast stain, 4D Streak plates (Quiz #2 - metrics)

5 18-MarRead 4D; Demo 10 Capsule; Review Staining Techniques & Colony Morphology (Quiz #3 - metrics)

6 25-Mar Practicum I

7 1-Apr Discuss Metabolism & Media; Read Demo 13, Inoculate Exp. 15, 17, 19, 20; & Begin 29

8 8-Apr Quiz #4 Lab Media; Read Tests 15, 17, 19, 20 and Demos 18, 21 & 22; Do 24; Cont. 29

15-Apr SPRING BREAK

9 22-Apr Read 24 and Demos 11, 12, 14, 25, 26, 27; Cont. 29

10 29-Apr Read and discuss Demos 30 - 35

11 6-May Quiz #5 Biochemicals, Review - ALL exercises to date!

12 13-May Practicum II / BREAK / Orientation, Begin Final Project 36

13 20-MayDisease Case Studies; Continue 36 Orientation and Progress (Quiz # 6 - metrics) (Water analysis paper due)

14 27-May Disease Case Studies; Continue 36

15 3-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

MicrobesSpring 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: http://youtu.be/X-w98KA8UqU

• Video on oil immersion: http://youtu.be/4ulrRzI_5Qc

• 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

10X 4X 40X

10X 10X 100X

10X 40X 400X

10X100X

(oil immersion)1,000X

2. Microscopy

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

AmoebaGiardia Trypanosoma

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 15

2. 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