Welcome to Micro Lab

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Welcome to Micro Lab!!!!

• TA Barb Honchak TAA Katie Conway
• Section E
• Email bmh43_at_dana.ucc.nau.edu
• Class web site http//www4.nau.edu/biology/bio205
  
• My open hours
• Tues. 4-5 Rm 125
• Wend. 4-5 Rm 124

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Today

• Orientation and Introduction
• Lab Rules
• Laboratory Protocol and safety
• Safety Quiz
• Required Laboratory Materials
• Lab Reports and Rubrics
• Mini Writing Assignment
• Syllabus
• Environmental Isolate
• Microscopy
• Care and Use
• Parts and function

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Lab Rules pg vii-ix in your manual

• Dress code and conduct
• No shorts-legs must be covered
• No open toed shoes
• Long hair must be pulled back
• You must wear a lab coat while in the lab
• You must come to class on time

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Protocol and Safetypg vii-ix in your manual

• Initial and sign handout
• Locate safety equipment in lab
• Locate disposal containers
• Safety quiz

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Required Lab Materialspage x in your manual

• Lab coat bookstore
• Sharpie bookstore
• Lab manual Kinko's
• Photo atlas bookstore
• Writing guide bookstore

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Mini Writing Assignments

• The purpose of these assignments are to help
  prepare you for your lab reports!!!!
• You have 4 mini writing assignments
• MWA1 is due next week

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Lab Reportspage xi in manual

• You have 3 lab reports
• You will have 2 attempts for the first two
  reports
• The final report you will only have one attempt
• Rubrics

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Syllabuspages i-v

• Take your syllabus out of your lab manual and
  tear it up!

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Microbiology Lab
Exercise 1

• Beginning the Environmental Isolate
• Introduction to the Compound Microscope

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ENVIRONMENTAL ISOLATE

• This is a semester long project and the subject
  of your final lab report (see 5-9)
• SO prepare to get friendly with your bacteria.
• Learn about media, Stains, Biochemical and
  Physiological test
• Learn to love Bergeys

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Solid Media are Prepared using agar

• -Agar A complex galactan (gelatine) which is
  widely used (in gel form) as a base for many
  kinds of solid and semi-solid microbiological
  medium.
• Agar is produced by many Rhodophyceae algae
  (red-brown algae) and is obtained commercially
  from e.g. Gelidium and Gracilaria spp

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Some Properties of Agar

• Once set, the agar is stable at temperatures up
  to 65C or higher.
• Few organisms are able to degrade agar although
  it does dehydrate.
• Agar is a transparent gelatine-like substance
  that is formed when a mixture of agar and water
  is heated to gt100C and then cooled gelling
  occurs at 40-45C.
• Growth media is prepared by adding nutrients,
  selective agents etc to agar usually before
  autoclaving.

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Our First Medium Trypticase Soy Agar (medium de
jour)

• -Trypticase Soy Agar (TSA) a general purpose
  medium, agar-based containing a pancreatic digest
  of casein (trypticase, tryptone), a papain digest
  of soybean meal (phytone), and NaCl (0.5 w/v).
• -Trypticase Soy Broth (TSB) is the same general
  purpose medium without agar base.

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Collecting Your Environmental Isolate Part 2

• Labeling a plate
• What is a control and why are they important
• A reference point in a controlled experiment in
  which a set of conditions does not vary
• Removing and handle sterile swabs
• Streaking a plate

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Collecting Your Environmental Isolate

• Collection procedures and location
• Mini Field Trip
• 10 min to collect. BE BACK BY ____
• Check your plates for label and incubate on the
  appropriate section shelf at RT

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Finally, to the Scopes!!!
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Introduction toMicroscopy using the Compound
Microscope

• The Olympus CH-2
• Phase Contrast Compound Microscope

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MICROSCOPY Several Methods and
their uses

• Bright-field
• The most common form of light microscopy.
  Extensively used for the visualization of
  microorganisms usually necessary to stain
  specimens for viewing.

Enterococcus
Anthrax spore stain
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MICROSCOPYSeveral Methods and their uses

• Dark-field Used for viewing live, unstained
  material. The specimen appears bright on a dark
  background. For low- or medium-power work.

Diatom
Rotifer
Cyanobacteria Nostoc
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MICROSCOPY Part 1 Several Methods
and their uses
Amoeba

• Phase contrast Microscopy used when a colorless
  specimen, which absorbs little light, (e.g. a
  non-pigmented living cell) is not clearly visible
  by bright-field microscopy

Bacteria
Rotifer
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MICROSCOPY Several Methods and
their uses

• Fluorescence microscopy
• Fluorochrome treated specimens (fluorescent
  stained) are irradiated with ultra-violet
  radiation and the light emitted forms the image
  of the specimen in a manner similar in principal
  to that in bright-field microscopy. The
  fluorescent scope is designed so that the
  specimen can be illuminated at one wavelength of
  light and observed by light emitted at a
  different wavelength.

Yersinnia pestis
Rhizopus rot-Black bread mold
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MICROSCOPY Several Methods and their uses

• Nomarski differential interference contrast
  microscopy a combination of light waves that are
  out of phase with each other and produces
  interference that alters the amplitude of the
  light waves. It produces high contrast images of
  unstained, transparent specimens in what appear
  to be three dimensions.

Heliozoan Actinomycies
Amoeba Nucleus and Vacuole
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MICROSCOPY Several Methods and their uses

• Electron microscopy microscopy in which an
  electron beam interacts with a specimen and
  contributes to an image of the object. Electron
  microscopy is used for examining viruses,
  macromolecules, and the ultra structure of cells.
  (electronically colored)

 Vibrio parahaemolyticus
Mixed Bacteria
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What Determinesthe Image you see?

• Magnification The extent to which the image of
  an object is larger than the object itself.
• Resolution The ability of a microscope to reveal
  fine detail in a specimen.
• Contrast The use of elements, such as colors,
  light, forms, or lines, in proximity to produce
  an intensified effect.
• We will go into more detail on these next week!

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Knowing your Microscope

• Parts and Maintenance

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All The Bits
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Bit By Bit
Ocular or eyepiece
Diopter adjustment
Observation tube clamping screw Do
Not Touch
Binocular head with observation tube
Arm
Revolving nosepiece
Objective lens 10x, 40x, 100x
Mechanical Stage with slide clip
Condenser
Condenser height adjustment knob
Fine focus knob
Iris diaphragm lever
Course focus adjustment
Phase-contrast condenser ?? (Should be
here)
Rheostat
Power switch
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THE MICROSCOPEParts Maintenance and Functions
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THE MICROSCOPEParts Maintenance and Functions

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THE MICROSCOPEParts Maintenance and Functions
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THE MICROSCOPEParts Maintenance and Functions
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THE MICROSCOPE its parts, their function,
maintenance and use
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THE MICROSCOPE its parts, their function,
maintenance and use
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THE MICROSCOPE its parts, their function,
maintenance and use
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Practice maintenance of scopes

• Clean lenses with lens paper and lens cleaner
• Clean condenser lens with lens paper and lens
  cleaner
• Clean stage with kimwipe and 70 alcohol (do not
  go over condenser lens with kimwipe!!!!!!!)
• Clean body with 70 alcohol and kimwipe