Chapter 2

PowerPoint® Lecture
Presentations prepared by
John Zamora
Middle Tennessee State
University
CHAPTER 2
Microbial Cell
Structure and
Function
© Pearson Education Limited 2015

I. Microscopy
• 2.1 Discovering Cell Structure: Light Microscopy
• 2.2 Improving Contrast in Light Microscopy
• 2.3 Imaging Cells in Three Dimensions
• 2.4 Probing Cell Structure: Electron Microscopy

2.1 Some Principles of Light Microscopy
• Compound light microscope uses visible light to

illuminate cells
• Many different types of light microscopy:

• Bright-field
• Phase-contrast
• Dark-field
• Fluorescence

Light Microscopy

• Bright-field scope
Specimens are visualized
because of differences in
contrast (density)
between specimen and
surroundings (Figure
2.2)

Figure 2.1b
Magnification Light path

100, 400, Visualized
1000 image
Two sets of lenses Eye
form the image
Objective lens and
Ocular lens
ocular lens 10
Intermediate image
(inverted from that
of the specimen)
Upper limit for

10, 40, or Objective lens
magnification is 100 (oil)
~2,000 Specimen
None Condenser lens
Light source
Total magnification = objective magnification  ocular magnification

© 2012 Pearson Education, Inc.
• Magnification: the ability to make an object larger
• Resolution: the ability to distinguish two adjacent

objects as separate and distinct
• Resolution is determined by the wavelength of light
used and numerical aperture of lens
• Limit of resolution for light microscope is about

0.2 μm

2.2 Improving Contrast in Light Microscopy
• Improving contrast results in a better final image
• Staining improves contrast

• Dyes are organic compounds that bind to specific
cellular materials
• Examples of common stains are methylene blue,

safranin, and crystal violet

Staining

Figure 2.3
I. Preparing a smear
Spread culture in thin Dry in air

film over slide
II. Heat fixing and staining
Pass slide through Flood slide with stain;

flame to heat fix rinse and dry
III. Microscopy
Slide Oil
Place drop of oil on slide;

examine with 100
objective lens
• Differential stains: the Gram stain
• Differential stains separate bacteria into groups
• The Gram stain is widely used in microbiology

(Figure 2.4a)
• Bacteria can be divided into two major groups:
gram-positive and gram-negative
• Gram-positive bacteria appear purple, and gram-negative

bacteria appear red after staining (Figure 2.4b)
Step 1 Flood the heat-fixed
smear with crystal
Result: violet for 1 min
All cells purple
Step 2 Add iodine solution

for 1 min
Result:
All cells
remain purple
Step 3 Decolorize with

alcohol briefly
Result: — about 20 sec
Gram-positive
cells are purple;
gram-negative
cells are colorless
Step 4 G- Counterstain with

Result: safranin for 1–2 min
Gram-positive
(G+) cells are purple; G+
gram-negative (G-) cells
are pink to red

• Phase-Contrast Microscopy
• Allows for the visualization of

live samples
• Invented in 1936 by Frits Zernike
• Phase ring amplifies differences
in the refractive index of cell and Bright-field Phase-
surroundings contrast
• Improves the contrast of a
sample without the use of a stain
• Resulting image is dark cells on
a light background (Figure 2.5)
Dark-field

• Dark-field microscopy
• Allows for the visualization of live samples
• Light reaches the specimen from the sides
• Light reaching the lens has been scattered by specimen
• Image appears light on a dark background (Figure 2.5)
• Excellent for observing motility

• Fluorescence microscopy
• Used to visualize specimens that fluoresce
• Emit light of one color when illuminated with another color
of light (Figure 2.6)
• Cells fluoresce naturally (autofluorescence) or after they

have been stained with a fluorescent dye such as DAPI
• Widely used in microbial ecology for enumerating

bacteria in natural samples

Figure 2.6
Cyanobacteria Cyanobacteria
E. Coli stained with DAPI

2.3 Imaging Cells in Three Dimensions
• Differential Interference Contrast (DIC) Nucleus
Microscopy (a form of light mscope)

• Uses a polarizer to create two distinct
beams of polarized light
• Gives structures such as endospores,

vacuoles, and granules a 3-D appearance
• Structures not visible using bright-field

microscopy are sometimes visible using
DIC
2.3 Imaging Cells in Three Dimensions
Confocal Scanning Laser Microscopy
(CSLM)
• Uses a computerized microscope
coupled with a laser source to
fluorescent microscope to generate
a 3-D image (Fig 2.8)
• Computer can focus the laser on
single layers of the specimen
• Different layers can then be
compiled for a three-dimensional
image
• Resolution is 0.1  m for CSLM
2.4 Electron Microscopy
• Electron microscopes use
electrons instead of photons Electron
source
to image cells and structures
(Figure 2.9)
Evacuated
• Two types of electron chamber
Sample
microscopes: port
• Transmission electron
microscopes (TEM)
Viewing
• Scanning electron screen
microscopes (SEM)
2.4 Electron Microscopy Cytoplasmic DNA
membrane Septum Cell wall (nucleoid)
Transmission Electron
Microscopy (TEM)
• Electromagnets function as
lenses
• System operates in a Thin section of dividing bacteria
vacuum
• High magnification and
resolution (0.2 nm)
• Enables visualization of
structures at the molecular
level (Figure 2.10a and b)
• Specimen must be very thin
(20–60 nm) and be stained
Negatively stained hemoglobin
2.4 Electron Microscopy
Scanning Electron Microscopy (SEM)

• Specimen is coated with a thin
film of heavy metal (e.g., gold)
• An electron beam scans the
object
• Scattered electrons are collected
by a detector and an image is
produced (Figure 2.10c)
• Even very large specimens can
be observed A single cell is about 0.75m wid
• Magnification range of 15–
100,000
Electron Microscopy

II. Cells of Bacteria and Archaea
• 2.5 Cell Morphology
• 2.6 Cell Size and the Significance of Being Small

2.5 Cell Morphology
• Morphology = cell shape

• Major cell morphologies (Figure 2.11)
• Coccus (pl. cocci): spherical or ovoid
• Rod or Bacil: cylindrical shape
• Spirillum: spiral shape
• Cells with unusual shapes

• Spirochetes, appendaged bacteria, and filamentous
bacteria
• Many variations on basic morphological types

Coccus Spirochete
Stalk Hypha
Rod Budding and
appendaged bacteria
Spirillum
Filamentous bacteria

2.5 Cell Morphology
• Morphology typically does not predict physiology,

ecology, phylogeny, etc. of a prokaryotic cell
• May be selective forces involved in setting the
morphology
• Optimization for nutrient uptake (small cells and those
with high surface-to-volume ratio)
• Swimming motility in viscous environments or near
surfaces (helical or spiral-shaped cells)
• Gliding motility (filamentous bacteria)
2.6 Cell Size and the Significance of Being
Small
• Size range for prokaryotes: 0.2 µm to >700 µm in
diameter
• Most cultured rod-shaped bacteria are between 0.5 and
4.0 µm wide and < 15 µm long
• Examples of very large prokaryotes
• Epulopiscium fishelsoni (Figure 2.12a)
• Thiomargarita namibiensis (Figure 2.12b)
• Size range for eukaryotic cells: 10 to >200 µm in

diameter

Examples of very large
prokaryotes
Epulopiscium fishelsoni
Thiomargarita
namibiensis

Small
• Surface-to-volume ratios, growth rates, and
evolution
• Advantages to being small (Figure 2.13)
• Small cells have more surface area relative to cell volume
than large cells (i.e., higher S/V)
• Support greater nutrient exchange per unit cell

volume
• Tend to grow faster than larger cells

© Pearson Education Limited 2015 Figure 2.13
Small
• Lower limits of cell size
• Cellular organisms <0.15 µm in diameter are unlikely
• Open oceans tend to contain small cells (0.2–0.4 µm in

diameter)

III. The Cytoplasmic Membrane and Transport
• 2.7 Membrane Structure
• 2.8 Membrane Functions
• 2.9 Nutrient Transport

2.7 Membrane Structure
• Cytoplasmic membrane
• Thin structure that surrounds the cell
• Vital barrier that separates cytoplasm from environment
• Highly selective permeable barrier; enables

concentration of specific metabolites and excretion of
waste products

• Composition of membranes
• General structure is phospholipid bilayer (Figure 2.14)
• Contain both hydrophobic and hydrophilic components
• Can exist in many different chemical forms as a result of

variation in the groups attached to the glycerol backbone
• Fatty acids point inward to form hydrophobic
environment; hydrophilic portions remain exposed to
external environment or the cytoplasm

Glycerol
Fatty acids
Phosphate
Ethanolamine
Hydrophilic
region
Hydrophobic
Fatty acids
region
Hydrophilic
region
Glycerophosphates
Fatty acids

Membrane Structure

• Cytoplasmic membrane (Figure 2.15)

• 8–10 nm wide
• Embedded proteins
• Stabilized by hydrogen bonds and hydrophobic

interactions
• Mg2+ and Ca2+ help stabilize membrane by forming ionic

bonds with negative charges on the phospholipids
• Somewhat fluid

Out
Phospholipids
Hydrophilic
groups
6–8 nm
Hydrophobic
groups
In
Integral
membrane
proteins Phospholipid
molecule

• Membrane proteins
• Outer surface of cytoplasmic membrane can interact with a
variety of proteins that bind substrates or process large
molecules for transport
• Inner surface of cytoplasmic membrane interacts with
proteins involved in energy-yielding reactions and other
important cellular functions
• Integral membrane proteins
• Firmly embedded in the membrane
• Peripheral membrane proteins
• One portion anchored in the membrane
2.7 The Cytoplasmic Membrane
• Membrane-Strengthening Agents
• Sterols
• Rigid, planar lipids found in eukaryotic membranes
Strengthen and stabilize membranes
• Hopanoids
• Structurally similar to sterols
• Present in membranes of many Bacteria

• Archaeal membranes
• Ether linkages in phospholipids of Archaea (Figure 2.16)
• Bacteria and Eukarya that have ester linkages in

phospholipids
• Archaeal lipids lack fatty acids; have isoprenes instead
• Major lipids are glycerol diethers and tetraethers

(Figure 2.17a and b)
• Can exist as lipid monolayers, bilayers, or mixture

(Figure 2.17)
Ester
Ether
Bacteria Archaea
Eukarya
Isoprene

Phytanyl
Major lipids are

CH3 groups
glycerol diethers
Glycerol diether Isoprene unit
Biphytanyl
and tetraethers
Can exist as lipid
monolayers,
Diglycerol tetraethers
bilayers, or mixture
Crenarchaeol
Out Out
Glycerophosphates
Phytanyl
Biphytanyl or
crenarchaeol
Membrane protein
In In
Lipid bilayer Lipid monolayer

Figure 2.17
2.8 Functions of the Cytoplasmic Membrane
• Permeability Barrier
• Polar and charged molecules must be transported
• Transport proteins accumulate solutes against the
concentration gradient
• Protein Anchor
• Holds transport proteins in place
• Energy Conservation
Permeability barrier: Protein anchor: Energy conservation:

Prevents leakage and functions Site of many proteins that Site of generation and use of the
as a gateway for transport of participate in transport, proton motive force
nutrients into, and wastes out bioenergetics, and chemotaxis
of, Education
© Pearson the cellLimited 2015
2.9 Nutrient Transport
• Carrier-Mediated Transport Systems

• Show saturation effect
Rate of solute entry

• Highly specific
Transporter saturated
with substrate
Transport
Simple diffusion

External concentration of solute
• Three major classes of transport systems in

prokaryotes (Figure 2.20)
• Simple transport
• Group translocation
• ABC system
• All require energy in some form, usually proton

motive force or ATP

Simple transport: Out In
Driven by the energy
in the proton motive
force
Transported
substance
Group translocation: P
Chemical modification
of the transported R~ P
substance driven by
phosphoenolpyruvate
1
2
ABC transporter:
Periplasmic binding 3
proteins are involved ATP ADP + Pi
and energy comes
from ATP.

• Three transport events are possible: uniport,
symport, and antiport
• Uniporters transport in one direction across the
membrane
• Symporters function as co-transporters
• Antiporters transport a molecule across the
membrane while simultaneously transporting another
molecule in the opposite direction

• Simple Transport:
• Lac Permease of Escherichia coli
• Lactose is transported into E. coli by the simple
transporter lac permease, a symporter
• Activity of lac permease is energy driven
• Other symporters, uniporters, and antiporters
Out
In
Sulfate Potassium Phosphate Sodium-proton Lac permease
symporter uniporter symporter antiporter (a symporter)
Group Translocation:
• The Phosphotransferase System in E. coli
• Type of group translocation: substance transported is
chemically modified during transport across the
membrane
• Best-studied system
• Moves glucose, fructose, and mannose
• Five proteins required
• Energy derived from phosphoenolpyruvate
Glucose
Out
Cytoplasmic
membrane
Nonspecific components Specific components

Enz
IIC Direction
PE of glucose
transport
Enz HPr Enz Enz
IIa IIb
Pyruvate
In
Direction of P transfer
Glucose 6–P
• ABC (ATP-binding cassette) systems (Figure 2.23)

• >200 different systems identified in prokaryotes
• Often involved in uptake of organic compounds (e.g., sugars,

amino acids), inorganic nutrients (e.g., sulfate, phosphate),
and trace metals
• Typically display high substrate specificity
• Gram-negatives employ periplasmic-binding proteins and

ATP-driven transport proteins
• Gram-positives employ substrate-binding proteins and

membrane transport proteins
IV. Cell Walls of Bacteria and Archaea
• 2.10 Peptidoglycan
• 2.11 LPS: The Outer Membrane
• 2.12 Archaeal Cell Walls
• Structural differences between cell walls of

gram-positive and gram-negative Bacteria are
responsible for differences in the Gram stain
reaction
Gram-negative
cell wall
• Two layers:
LPS and
peptidoglycan
Gram-positive
cell wall
• One layer:
peptidoglycan

2.10 Peptidoglycan
• Peptidoglycan (Figure 2.25)

• Rigid layer that provides strength to cell wall
• Polysaccharide composed of:

• N-acetylglucosamine and N-acetylmuramic acid
• Amino acids
• Lysine or diaminopimelic acid (DAP)
• Cross-linked differently in gram-negative bacteria and

gram-positive bacteria (Figure 2.26)

N-Acetylglucosamine ( G ) N-Acetylmuramic acid ( M )
Glycan tetrapeptide
N-Acetyl
group
Lysozyme-
sensitive
Peptide
bond
cross-links
L-Alanine
D-Glutamic acid
Diaminopimelic
acid
D-Alanine

Polysaccharide
backbone
Interbridge
Peptides
Escherichia coli
(gram-negative)
Staphylococcus aureus
(gram-positive)

• Gram-positive cell walls
(Figure 2.27)
Peptidoglycan
• Can contain up to 90%

cable
peptidoglycan
• Common to have teichoic

acids (acidic substances) Wall-associated Teichoic acid Peptidoglycan Lipoteichoic
embedded in their cell wall protein acid
• Lipoteichoic acids:
teichoic acids covalently
bound to membrane lipids
Cytoplasmic membrane

2.10 Peptidoglycan
• Prokaryotes that lack cell walls

• Mycoplasmas
• Group of pathogenic bacteria
• Thermoplasma
• Species of Archaea
Acid-Fast Stain (Ziehl–Neelsen stain)

2.11 LPS: The Outer Membrane
• Total cell wall contains ~10% peptidoglycan

• Most of cell wall composed of outer membrane,
aka lipopolysaccharide (LPS) layer
• LPS consists of lipid A, core polysaccharide and O-
polysaccharide (Figure 2.28)
• LPS replaces most of phospholipids in outer half of outer
membrane (Figure 2.29)
• Endotoxin: the toxic component of LPS

O-specific Core polysaccharide
polysaccharide
Lipid A Protein Out
Lipopolysaccharide
(LPS)
Porin
Outer 8 nm
membrane
Cell
wall
Phospholipid
Peptidoglycan
Periplasm
Lipoprotein
Cytoplasmic
membrane
In
Outer membrane
Periplasm
Cytoplasmic
membrane

2.11 LPS: The Outer Membrane
• Periplasm: space located between cytoplasmic

and outer membranes (Figure 2.29)
• ~15 nm wide
• Contents have gel-like consistency
• Houses many proteins
• Porins: channels for movement of hydrophilic

low-molecular-weight substances (Figure 2.29c)

2.12 Archaeal Cell Walls
• No peptidoglycan
• Typically no outer membrane
• Pseudomurein
• Polysaccharide similar to peptidoglycan (Figure 2.30)
• Composed of N-acetylglucosamine and N-

acetylalosaminuronic acid
• Found in cell walls of certain methanogenic Archaea
• Cell walls of some Archaea lack pseudomurein

N--Acetyltalosaminuronic
acid( T )
Lysozyme-insensitive
N-Acetyl
N-Acetylglucosamine( G ) group
𝛃(1,3)
Peptide
cross-links
T G

2.12 Archaeal Cell Walls
• S-Layers
• Most common cell wall type
among Archaea
• Consist of protein or
glycoprotein
• Paracrystalline structure

V. Other Cell Surface Structures and Inclusions
• 2.13 Cell Surface Structures
• 2.14 Cell Inclusions
• 2.15 Gas Vesicles
• 2.16 Endospores

2.13 Cell Surface Structures
Acinetobacter
• Capsules and Slime Layers
– Polysaccharide layers
• May be thick or thin, rigid or
flexible
– Assist in attachment to
surfaces Rhodobacter capsulatus
Cell Capsule
– Protect against phagocytosis
– Resist desiccation
Rhizobium trifolii
• Fimbriae
• Filamentous protein structures
• Enable organisms to stick to surfaces or form pellicles
Flagella
Fimbriae

• Pili
• Filamentous protein structures (Figure 2.34)
• Typically longer than fimbriae
• Assist in surface attachment
• Facilitate genetic exchange between cells
(conjugation)
• Type IV pili involved in twitching motility
Virus-
covered
pilus

2.14 Cell Inclusions
• Carbon storage polymers

• Poly-β-hydroxybutyric acid (PHB): lipid (Figure 2.35)
• Glycogen: glucose polymer
• Polyphosphates: accumulations of inorganic phosphate
Sulfur globules: composed of elemental sulfur
• Carbonate minerals: composed of barium, strontium, and
magnesium (Figure 2.37)
• Magnetosomes: magnetic storage inclusions
• Carboxysomes: contain enzymes involved in CO2 fixation

β-carbon
Polyhydroxyalkanoate

Figure 3.27
Polyphosphate
Magnetotactic bacteria
Sulfur

Magnetosomes
Cyanobacterium Gleomargarita containing
granules of the mineral bensonite
2.15 Gas Vesicles
• Gas Vesicles
• Confer buoyancy in planktonic
cells
• Spindle-shaped, gas-filled
structures made of protein Anabaena
• Gas vesicle impermeable to

water

Microcyctis
2.15 Gas Vesicles Ribs
• Molecular Structure of
Gas Vesicles
• Gas vesicles are
composed of two
proteins: GvpA and
GvpA
GvpC
• Function by decreasing
GvpC
cell density

2.16 Endospores
• Endospores
• Highly differentiated cells resistant to heat, harsh
chemicals, and radiation
• “Dormant” stage of bacterial life cycle
• Ideal for dispersal via wind, water, or animal gut
• Present only in some gram-positive bacteria
Terminal Subterminal Central

spores
© Pearson Education Limited 2015 spores spores
Vegetative cell
Developing
endospore
Sporulating cell
Mature endospore

2.16 Endospores
Exosporium
Spore coat
• Endospore Structure Core wall
• Structurally complex Cortex
DNA
• Contains dipicolinic acid
• Enriched in Ca2+
• Core contains small acid-

soluble proteins (SASPs)

• The Sporulation Process
– Complex series of events
– Genetically directed
Coat
Spore coat, Ca2

Free endospore Maturation, uptake, SASPs,
cell lysis dipicolinic acid
Stage VI, VII

Growth Germination Stage V
Cortex
Vegetative Sporulation Cell wall
cycle stages Cytoplasmic
membrane
Cell Asymmetric
division cell division;
commitment Cortex Stage IV
to sporulation, formation
Stage I
Prespore
Septum
Engulfment
Mother cell
© Pearson Education Limited 2015 Stage II Stage III

VI. Microbial Locomotion
• 2.17 Flagella and Swimming Motility
• 2.18 Gliding Motility
• 2.19 Chemotaxis and Other Taxes

2.17 Flagella and Swimming Motility
• Flagella: structure that assists in swimming

• Different arrangements: peritrichous, polar,
lophotrichous (Figure 2.48)
• Helical in shape

The Prokaryotic Flagellum

• Flagellar structure of Bacteria

• Consists of several components (Figure 2.51)
• Filament composed of flagellin
• Move by rotation
• Flagellar structure of Archaea

• Half the diameter of bacterial flagella
• Composed of several different proteins
• Move by rotation
15–20 nm
L
Filament P
Flagellin MS
Hook
Outer
membrane
(LPS)
L
Ring
Rod
P
Periplasm Ring
Peptidoglycan
+ + ++ + + ++
MS Ring
Basal
body
−−−− C Ring −−−−
Cytoplasmic Mot protein Fli proteins Mot protein

membrane (motor switch)
45 nm
Rod
MS Ring
Mot
protein
C Ring

• Flagellar synthesis
• Several genes are required for flagellar synthesis and motility
• MS ring is made first
• Other proteins and hook are made next
• Filament grows from tip Filament

synthesis
Hook-
Late hook filament
Outer Cap junction
Early hook
membrane Filament
Motor (Mot)
MS/C ring proteins P ring L ring
Peptidoglycan Cytoplasmic membrane

• Flagella increase or decrease rotational speed in

relation to strength of the proton motive force
• Differences in swimming motions (Figure 2.54)

• Peritrichously flagellated cells move slowly in a straight
line
• Polarly flagellated cells move more rapidly and typically

spin around

Tumble − flagella
pushed apart
(CW rotation)
Bundled
flagella
(CCW rotation)
Flagella bundled
(CCW rotation)
Peritrichous
Reversible flagella
CCW rotation CW rotation
Unidirectional flagella
Cell
stops,
CW rotation reorients
CW rotation
Polar

2.18 Gliding Motility
• Gliding motility
• Flagella-independent motility (Figure 2.56)
• Slower and smoother than swimming
• Movement typically occurs along long axis of cell
• Requires surface contact
• Mechanisms
• Excretion of polysaccharide slime
• Type IV pili
• Gliding-specific proteins
In
Cytoplasmic
membrane
Peptidoglycan
Outer
membrane
Out Glide proteins

Movement of outer Surface
Movement of cell membrane glide proteins

2.19 Chemotaxis and Other Taxes
• Taxis: directed movement in response to chemical

or physical gradients
• Chemotaxis: response to chemicals
• Phototaxis: response to light
• Aerotaxis: response to oxygen
• Osmotaxis: response to ionic strength
• Hydrotaxis: response to water

• Chemotaxis
• Best studied in E. coli
• Bacteria respond to temporal, not spatial, difference in

chemical concentration
• “Run and tumble” behavior (Figure 2.57)
• Attractants and receptors sensed by chemoreceptors

Tumble Attractant
Tumble
Run
Run
No attractant present: Random Attractant present: Directed

movement movement

• Measuring chemotaxis (Figure 2.58)

• Measured by inserting a capillary tube containing an
attractant or a repellent in a medium of motile bacteria
• Can also be seen under a microscope

VII. Eukaryotic Microbial Cells
• 2.20 The Nucleus and Cell Division
• 2.21 Mitochondria, Hydrogenosomes, and

Chloroplast
• 2.22 Other Major Eukaryotic Cell Structures

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PowerPoint® Lecture

© Pearson Education Limited 2015

• 2.1 Discovering Cell Structure: Light Microscopy

• 2.2 Improving Contrast in Light Microscopy

• 2.3 Imaging Cells in Three Dimensions

• 2.4 Probing Cell Structure: Electron Microscopy

© Pearson Education Limited 2015

• Compound light microscope uses visible light to

• Many different types of light microscopy:

© Pearson Education Limited 2015

© Pearson Education Limited 2015

© Pearson Education Limited 2015

Magnification Light path

Upper limit for

Total magnification = objective magnification  ocular magnification

• Magnification: the ability to make an object larger

• Resolution: the ability to distinguish two adjacent

• Limit of resolution for light microscope is about

© Pearson Education Limited 2015

• Improving contrast results in a better final image

• Staining improves contrast

• Examples of common stains are methylene blue,

© Pearson Education Limited 2015

© Pearson Education Limited 2015

Spread culture in thin Dry in air

II. Heat fixing and staining

Pass slide through Flood slide with stain;

Place drop of oil on slide;

• Differential stains: the Gram stain

• Differential stains separate bacteria into groups

• The Gram stain is widely used in microbiology

gram-positive and gram-negative

• Gram-positive bacteria appear purple, and gram-negative

Step 2 Add iodine solution

Step 3 Decolorize with

Step 4 G- Counterstain with

© Pearson Education Limited 2015

• Allows for the visualization of

© Pearson Education Limited 2015

• Light reaches the specimen from the sides

• Light reaching the lens has been scattered by specimen

• Image appears light on a dark background (Figure 2.5)

• Excellent for observing motility

© Pearson Education Limited 2015

• Cells fluoresce naturally (autofluorescence) or after they

• Widely used in microbial ecology for enumerating

© Pearson Education Limited 2015

E. Coli stained with DAPI

• Differential Interference Contrast (DIC) Nucleus

Microscopy (a form of light mscope)

• Gives structures such as endospores,

• Structures not visible using bright-field

Scanning Electron Microscopy (SEM)

© Pearson Education Limited 2015

• 2.5 Cell Morphology

• 2.6 Cell Size and the Significance of Being Small

© Pearson Education Limited 2015

• Morphology = cell shape

• Cells with unusual shapes