Welcome to
Microbiology Lab
Tue 6:00-9:10pm
Instructor: Huma Malik
Email: Huma.Malik@rccd.edu
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
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
• You will receive one grade for Microbiology lecture
and lab:
• Lab = 30%
• Lecture = 70%
Wk
Date
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-Mar
4
11-Mar
8A Motility Hanging drop, 9 Acid Fast stain, 4D Streak plates
(Quiz #2 - metrics)
5
18-Mar
Read 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
Lab Assignment
7 Endospore stain, Read 8B motility tall, 4 (23) Oxygen Req's, 16 Catalase Test
(Quiz #1 - metrics)
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-May
Disease 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
16
Disease Case Studies; 36 Final Project Due - you should be checked out by June 5
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
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
10X
100X
(oil immersion)
1,000X
2. Microscopy
Where Are Microorganisms Found and How Do We Study Them?
Lab 2
• Bacterial cells have certain shapes and arrangements
Bacteria - 1000x
Coccus (round)
Rod (bacillus)
Spirochete
Single
Pair
Single
Tetrad
Chain
Chain
Cluster
(masses)
______________________________________________________________________________
2. Microscopy
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
Giardia
Amoeba
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
3. Ubiquity of Microbes
• 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