LABORATORY SKILLS
MIC-240
Dr. Kahkashan Perveen
Bulding. 4, Floor 1, Room 415
E-mail: kperveen@ksu.edu.sa
Dr. K. Perveen. MIC 240, K.S.U
• Tests
and Exams:
REPORTS
10 %
QUIZ
20 %
ASSIGNMENT
15 %
CLASS PERFORMANCE
10 %
FINAL EXAM
45%
Dr. K. Perveen. MIC 240, K.S.U
Part 1:Microbiology
Laboratory, Organization and
Management
a. Laboratory Safety Rules
b. Apparatus and
Equipments
Dr. K. Perveen. MIC 240, K.S.U
Laboratory Safety Rules
1. Always wear a laboratory coat.
2. Put nothing in mouth which may have
come in contact with infectious
material.
3. Eating and drinking in the laboratory
are not permitted at any time.
Dr. K. Perveen. MIC 240, K.S.U
4. Keep
your workspace free of all
unnecessary materials.
5. Never pipette by mouth. Use the
safety pipetting devices which are
provided.
Dr. K. Perveen. MIC 240, K.S.U
6. Throw
off used pipettes
appropriate containers.
in
the
7. Any infectious material which may
accidentally fall from pipettes to the
laboratory bench or floor should be
covered with a disinfectant and
reported
to
any
instructor
immediately.
Dr. K. Perveen. MIC 240, K.S.U
8.
Any spilled or broken containers of
culture material should be covered
with disinfectant and then brought to
the attention of an instructor.
Dr. K. Perveen. MIC 240, K.S.U
9. Replace caps on reagents, solution
bottles, and bacterial cultures after.
10. Do not open Petri dishes in the lab
unless absolutely necessary.
11. Take care of the microscope.
12. Label everything clearly.
Dr. K. Perveen. MIC 240, K.S.U
13. When finished for the day, dispose of
all used glassware and cultures in the
appropriate
containers,
clear
workbench and wash the top with a
disinfectant.
Dr. K. Perveen. MIC 240, K.S.U
14. Make sure all burners are turned
off at the end of the laboratory
period.
15. Take off your gloves when you
leave the laboratory.
Dr. K. Perveen. MIC 240, K.S.U
16. Wash hands thoroughly with soap
and water
laboratory.
before
leaving
the
17. Familiarize yourself with the
location of safety equipment in the
laboratory.
Dr. K. Perveen. MIC 240, K.S.U
Laboratory Safety Equipment
Eyewash and shower
Sinks
Fire Extinguisher
First Aid Kit
Emergency Gas Valve
Dr. K. Perveen. MIC 240, K.S.U
Apparatus and
Equipments
Dr. K. Perveen. MIC 240, K.S.U
1. Microscope
Used to observe very small organisms
Dr. K. Perveen. MIC 240, K.S.U
2. Autoclave
It is a wet/ type sterilizer.
Used to sterilize culture media, glassware
etc.
Usually it operates at 15 lb/sq. inch steam
pressure (121.5 o C) for 30 min.
Dr. K. Perveen. MIC 240, K.S.U
3. Incubator
Provide suitable temperature for the
growth of organism
Dr. K. Perveen. MIC 240, K.S.U
4. Hot Air Oven
It is a dry air type sterilizer.
It is used for sterilizing laboratory glass
ware.
It operates at a temperature of 160 to 180
oC for one and a half hour.
Dr. K. Perveen. MIC 240, K.S.U
5. Laminar Air Flow
It is a chamber which provide microbe free
environment.
It is transfer of media
for culturing bacteria or fungi
or any microbes.
Dr. K. Perveen. MIC 240, K.S.U
6. Centrifuge
It spins liquid samples to separate their
components
Dr. K. Perveen. MIC 240, K.S.U
7. Spectrophotometer
uses wavelength to determine the
concentration of a compound or particles in a
solution or suspension.
Dr. K. Perveen. MIC 240, K.S.U
8. Micropipette
Pipettes are used to accurately measure
and dispense small volumes of liquid.
Dr. K. Perveen. MIC 240, K.S.U
9. Inoculating loops &
Inoculating needles
Used for inoculating microbes in the liquid
media & solid media
Dr. K. Perveen. MIC 240, K.S.U
10. Bunsen burner
Source of flame
Dr. K. Perveen. MIC 240, K.S.U
11. Slide
Glass support for specimens
Dr. K. Perveen. MIC 240, K.S.U
12.Cover Slip
Glass cover for specimens
Dr. K. Perveen. MIC 240, K.S.U
13. Petri Dishes
Dr. K. Perveen. MIC 240, K.S.U
14. Gram Staining Kits
Dr. K. Perveen. MIC 240, K.S.U
15. Growth Media/Culture
Media
Dr. K. Perveen. MIC 240, K.S.U
16. Disinfectant
Clorox Bleach, diluted to 5-10% is the best
cleaning agent for labs.
Dr. K. Perveen. MIC 240, K.S.U
Microscopy
Dr. K. Perveen. MIC 240, K.S.U
Compound Microscope
Instrument
for
observing
small objects
Magnify
images up to
2000X their
size
Dr. K. Perveen. MIC 240, K.S.U
Different parts of
a microscope
Dr. K. Perveen. MIC 240, K.S.U
Revolving
nosepiece
Eyepiece
Clip
Body tube
Coarse
adjustment
Fine adjustment
Condenser
Arm
Iris diaphragm
Stage
Objective
Condenser
control knob
Mirror
Base
Dr. K. Perveen. MIC 240, K.S.U
Microscope
Dr. K. Perveen. MIC 240, K.S.U
Name
Characteristics
Magnifying power
Scanning power
shortest objective, red stripe
4X
Low power
next shortest, yellow stripe
10 X
High-dry power
intermediate length, blue
40, 43 or 45 X
stripe
Oil immersion
longest, black stripe
Dr. K. Perveen. MIC 240, K.S.U
100 X
Resolution
It is the ability to differentiate two objects
close together as being separate.
Dr. K. Perveen. MIC 240, K.S.U
Magnification
Total magnification: To calculate
the total magnification of any
specimen being viewed multiply
the power of the eye piece (ocular
lens) by the power of the objective
lens being used.
Dr. K. Perveen. MIC 240, K.S.U
Example
If
the eye piece magnifies = 10X
the objective lens magnifies = 40X,
then 10 x 40 = 400X (total
magnification)
Dr. K. Perveen. MIC 240, K.S.U
Care of the microscope
Mentioned in the text
Dr. K. Perveen. MIC 240, K.S.U
Types of Microscopy
Brightfield Microscope: This
microscope is used to observe
nonviable, stained preparation.
Darkfield Microscope
Phase-Contrast Microscope: The
unstained microorganisms are
observed easily.
Dr. K. Perveen. MIC 240, K.S.U
•
•
•
•
Fluorescent Microscope: This microscope
is used to observe the specimens that are
chemically treated with a fluorescent dye.
The specimens are illuminated with an
ultraviolet light.
This microscope is used for the detection of
antigen-antibody reactions.
The fluorescent portion of the dye becomes
visible against the black background.
Dr. K. Perveen. MIC 240, K.S.U
Fluorescent image of cultured rat-brain
cells. Living cells stain with calcein (left)
and dead cells stain with propidium iodide
(right).
Dr. K. Perveen. MIC 240, K.S.U
Electron Microscope: It
produces an electronicallymagnified image of a
specimen for detailed
observation.
It is used to observe
submicroscopic cellular
particles as well as viral
agents.
The specimen is illuminated
by a beam of electrons
rather than light.
Dr. K. Perveen. MIC 240, K.S.U
How to Make a Wet Mount
Water drop
Cover slip
Object/specimen
Slide
Dr. K. Perveen. MIC 240, K.S.U
Technique for Adding a Stain
when making a Wet Mount
Dr. K. Perveen. MIC 240, K.S.U
Exercise 1: Prepare the wet mount of the
yeast from the culture provided to u.
Observe the slide under low and high
power and draw the diagram in the result
sheet.
Dr. K. Perveen. MIC 240, K.S.U
Observations and Results
Exercise 1a
Name:___________________________________________
I.D No.___________________________________________
Date:__________________
Group:________________
Observation
Low Power (10x)
Draw yeast
structure
Magnification
Dr. K. Perveen. MIC 240, K.S.U
High Power (40x)
Dr. K. Perveen. MIC 240, K.S.U
Thanks
Dr. K. Perveen. MIC 240, K.S.U