Lecture 3

Microbial Cell Biology
Lecture 3 Lani Manahan-Suyom
HOW DO WE OBSERVE MICROBIAL CELLS?
Microscopy
DEFINITION OF TERMS
Resolution is the ability of the lenses to distinguish two points
Function of the wavelength used and the numerical aperture
Shorter wavelengths of light provide greater resolution
A microscope with a resolving power of 0.2 m (200 nm)
Numerical aperture light gathering ability of the microscope Refractive index is the light-bending ability of a medium
lecture 3 MICROBIAL CELL BIOLOGY 3
THE USE OF OIL IMMERSION

Immersion oil is used to keep light from bending
lecture 3 MICROBIAL CELL BIOLOGY
TYPES OF MICROSCOPES
Dark objects are visible against a bright background Light reflected off the specimen does not enter the objective lens.
Light objects are visible against a dark background Light reflected off the specimen enters the objective lens.
PHASE CONTRAST MICROSCOPY

Accentuates diffraction of the light that passes through a specimen
DIFFERENTIAL INTERFERENCE MICROSCOPY

Accentuates diffraction of the light that passes through a specimen; uses two beams of light
FLUORESCENCE MICROSCOPY
Uses UV light
Fluorescent substances absorb UV light and emit visible light
Cells may be stained with fluorescent dyes (fluorochromes)

CONFOCAL MICROSCOPY
Uses fluorochromes and a laser light The laser illuminates each plane in a specimen to produce a 3-D image
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ELECTRON MICROSCOPY
Uses electrons instead of light The shorter wavelength of electrons gives greater resolution
TRANSMISSION ELECTRON MICROSCOPY (TEM)

Ultrathin sections of specimens. Beam passes through specimen, then an electromagnetic lens, to a screen or film. Specimens may be stained with heavy metal salts.
10,000-100,000; resolution 2.5 nm
15
SCANNING ELECTRON MICROSCOPY

An electron gun produces a beam of electrons that scans the surface of a whole specimen
Secondary electrons emitted from the specimen produce the image
1000-10,000; resolution 20 nm
17
SCANNING TUNNELING MICROSCOPY

Scanning tunneling microscopy uses a metal probe to scan a specimen
Resolution 1/100 of an atom
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ATOMIC FORCE MICROSCOPY

Atomic force microscopy uses a metal and diamond probe inserted into the specimen Produces 3-D images Metal coat not needed
19
Looking into cell details

Size, shape and arrangement
Cell Morphology (shape)

SHAPE BASED ON Genetics
something to do with being monomorphic each bacteria has its own shape
Environmental Factors
pleiomorphism due to absence of cell wall
ex. Mycoplasma (rigidity: ergosterol)
CELL DIVISION AND SHAPES: COCCI
DIPLOCOCCI STREPTOCOCCI
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TETRAD
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STAPHYLOCOCCI
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CELL DIVISION AND SHAPES: SPIRALS
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OTHER SHAPES OF BACTERIA
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UNUSUAL SHAPES
Most bacteria are monomorphic A few are pleomorphic
Stella sp.
Haloarcula sp
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VARIOUS ARANGEMENTS
Pairs: diplococci, diplobacilli Clusters: staphylococci Chains: streptococci, streptobacilli
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CELL DIVISION AND SHAPES: BACILLI

STREPTOBACILLI
BACILLI
COCCOBACILLI DIPLOBACILLI
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Importance of cell size?
Size affects surface to volume ratio Nutrients and wastes can pass readily in and out of a smaller cell than larger cells accelerating cell growth
Bacteria
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Thiomargarita namibiensis
Epulopiscium fishelsoni
PROKARYOTIC CELL ORGANIZATION
34
TYPICAL BACTERIAL CELL
35
FUNCTIONS OF PROKARYOTIC STRUCTURES

Cell Wall Gives bacteria shape and protection from lysis in dilute solutions
Plasma Membrane
Selectively permeable barrier, mechanical boundary of cell, nutrient and waste transport, location of many metabolic processes (respiration, photosynthesis), detection of environmental cues for chemotaxis
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Capsules and Slime Layers Resistance to phagocytosis, adherence to surfaces Attachment to surfaces, bacterial mating Movement
Survival under harsh environmental conditions
Fimbriae and Pili
Flagella
Endospore

Nucleoid
Periplasmic Space
Ribosomes Gas Vacuole Inclusion Bodies
Localization of Genetic Material (DNA)

Contains hydrolytic enzymes and binding proteins for nutrient processing and uptake
Protein synthesis Buoyancy for floating in aquatic environments Storage of carbon, phosphate and other substances
THE PROKARYOTIC CELL WALL
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Cell Walls
Cell wall is a structure that completely surrounds the cell protoplast. (Almost) all bacteria have a cell wall.
IMPORTANCE OF CELL WALLS FOR BACTERIA

Gives shape and protection from osmotic lysis
May contribute to pathogenicity Protection from toxic substances Site of action of several antibiotics
CHRISTIAN GRAM AND HIS STAIN

Christian Gram developed a Stain (1884)
Some are colored red/pink Some are colored blue/violet
Divided the microbial world into 2: positive and negative

Reaction to the staining process
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Profile of the bacterial cell envelope
Gram-positive cell wall is thick homogeneous monolayer Gram-negative cell wall is thin heterogeneous multilayer Periplasmic space: space between the plasma membrane and the outer membrane (Gram negative bacteria)
Loose PG and some proteins For Gram + counterpart are exoenzymes
DIFFERENTIATING GRAM POSITIVE AND NEGATIVE WALLS
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Chemical nature of bacterial cell walls

Bacterial cell walls always contain murein, which is a type of peptidoglycan
Chemical nature of murein accounts for the function of the cell wall Murein is only found in the cell walls of bacteria
E. coli peptidoglycan

Peptidoglycan is made up of 2 amino sugars N-acetyl-glucosamine = G N- acetylmuramic acid = M 4 amino acids L-alanine = L-ala D-glutamic acid = D-glu diaminopimelic acid = DAP D-alanine = D-ala
Gram-negative murein. Murein is a polymer of the peptidoglycan subunit. The sugars form the glycan backbone (G-M-G-M-etc.) and the amino acids comprise the peptide side chains of the molecule.

Penicillin prevents formation of this Interpeptide bond
Lysozyme breaks this glycoside bond between M and G
Gram-negative murein showing the sites of action of the antibiotic penicillin and the enzyme lysozyme
Gram-positive murein has a thicker glycan a backbone and there are interpeptide bridges that join amino acid side chains together.
Penicillin blocks the Insertion of the interpeptide bridge
Lysozyme breaks the glycoside bond between M and G
Gram-positive murein showing the sites of action of the antibiotic penicillin and the enzyme lysozyme
Other characteristics of bacterial cell walls

Gram-positive cell walls contain teichoic acids
Teichoic acids are thought to stabilize the Gram positive cell wall and may be used in adherence.
TEICHOIC ACIDS
Teichoic acids:
Lipoteichoic acid links to plasma membrane Wall teichoic acid links to peptidoglycan
May regulate movement of cations Polysaccharides provide antigenic variation


Gram-negative cell walls include an outer membrane
CHARACTERISTICS
Lipopolysaccharides, lipoproteins, phospholipids. Forms the periplasm between the outer membrane and the plasma membrane.
Protection from phagocytes, complement, antibiotics.

O polysaccharide antigen, e.g., E. coli O157:H7. Lipid A is an endotoxin. Porins (proteins) form channels through membrane

Outer membrane of Gram-negatives has two important properties 1. It protects the cells from permeability by many substances including penicillin and lysozyme. 2. It is the location of lipopolysaccharide (endotoxin) which is toxic for animals.
Table: Correlation of the Gram stain with properties of bacterial cell walls
Property Thickness of wall Number of layers Peptidoglycan (murein) content Teichoic acids in wall Protein/lipoprotein content Lipopolysaccharide content Sensitivity to penicillin Gram-positive thick (20-80 nm) 1 >50% present 0-3% 0 sensitive Gram-negative thin (10 nm) 2-3 10-20% absent >50% 13 resistant resistant
Sensitivity to lysozyme sensitive
GRAM POSITIVE BACTERIA

react with the Gram stain to appear purple, whereas Gram negative bacteria don't react with the stain
cell wall of gram positive bacteria consists of several layer of peptidoglycan units In addition, the cell wall of gram positive bacteria contains teichoic acid, which consists of glycerol or ribitol and phosphate
GRAM NEGATIVE BACTERIA

only single layer of peptidoglycan, connected to a lipoprotein unit (Brauns lipoprotein) Outer membrane -lipopolysaccharide Lipid A portion two sugar units connected to a hydroxy fatty acid referred as an endotoxin since it is toxic in a host's bloodstream or gastrointestinal tract O-antigen subunit consists of sugar molecules the primary site recognized by antibodies
THE PRINCIPLE BEHIND GRAM STAINING

Points to Ponder:
1.
2.
Why are Blue-stained cells termed as Gram Positive and Redstained cells Gram negative and not the other way around?
As cells age, what properties do they loose? Gram positivity or Gram negativity?
Gram Staining Procedure

1. crystal violet. Primary stain. Both gram-positive and gram-negative bacteria become directly stained and appear purple after this step. gram's iodine solution. Mordant. This allows the stain to be retained better by forming an insoluble crystal violet-iodine complex. Both gram-positive and gram-negative bacteria remain purple after this step. ethyl alcohol. Decolorizer. This is the differential step. Gram-positive bacteria retain the crystal violet-iodine complex while gram-negative are decolorized. safranin (also a basic dye). counterstain Since the gram-positive bacteria are already stained purple, they are not affected by the counterstain. Gramnegative bacteria, that are now colorless, become directly stained by safranin. 2.
3.
4.
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Principle:
Gram-positive and gram-negative bacteria stain differently because of fundamental differences in the structure of their cell walls
gram-positive cell wall have thick layers of peptidoglycan (60-90%) Low lipid content
gram-negative cell wall contains a much thinner, single layer of peptidoglycan (10-20%) only two or three layers thick. high lipid content (outer membrane)
Effect of solvent/decolorizer, (ethanol)

dehydrates peptidogyclan layer dissolves the lipid layer from the gram-negative cells.
The removal of the lipid layer enhances the leaching of the primary stain from the cells into the surrounding solvent. Dehydration causes closing the pores as the cell wall shrinks during dehydration.
WHEN GRAM STAIN IS NOT APPLICABLE

Mycoplasmas
Lack cell walls Sterols in plasma membrane
Archaea
Wall-less, or Walls of pseudomurein (lack NAM and D amino acids)
THE CELL WALL AND OSMOTIC PROTECTION

Recall: Osmosis
Solutes are much more concentrated in bacterial cytoplasm than in most habitats
Water will move in Without cell wall: cell will swell and lyze
Importance: treatment with penicillin or lysozyme

Lysozyme: attacks PG by hydrolyzing the bonds that connect NAM and NAG Penicillin: inhibits PG synthesis
PROKARYOTIC CELL MEMBRANES

an absolute requirement for all living organisms
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THE PLASMA MEMBRANE

Contains both proteins and lipids (proportions vary widely) Lipids are amphipathic (with polar and nonpolar ends)
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WHATS UNIQUELY THEIRS

BACTERIA
Lacks sterols (cholesterols) instead hopanoids to stabilize bacterial membranes
ARCHAEBACTERIA
Have a monolayer with lipid molecules spanning the whole membrane (to be discussed later)
THE FLUID MOSAIC MODEL

By S. Jonathan Singer and Garth Nicholson Distinguish between two types of membrane proteins Peripheral proteins Loosely connected to the membranes and can easily be removed Soluble in aqueous solutions 20-30% of total membrane proteins Integral proteins (amphipatic) 70-80% of total membrane proteins Not easily extracted from membranes Insoluble in aqueous solutions when freed of lipids
THE FLUID MOSAIC MODEL (Singer and Nicholson)
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LIPID COMPONENTS OF EUBACTERIA AND ARCHAEA

EUBACTERIA
Ester linkage Weaker linkage
ARCHAEBACTERIA
Ether linkage Stronger linkage
lecture 3 MICROBIAL CELL BIOLOGY POINT OF LINKAGE = POINT OF BREAKDOWN
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TRANSPORT SYSTEMS IN MEMBRANES

Simple diffusion: Movement of a solute from an area of high concentration to an area of low concentration
Facilitative diffusion: Solute combines with a transporter protein in the membrane Osmosis
Movement of water across a selectively permeable membrane from an area of high water concentration to an area of lower water Osmotic pressure: The pressure needed to stop the movement of water across the membrane
HOW DO MATERIALS MOVE THROUGH THEIR MEMBRANES?
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HOW DO MATERIALS MOVE THROUGH THEIR MEMBRANES?
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PHOTOSYNTHETIC MEMBRANES: chlorosomes

may be part of the PM or close to it
Example: Chlorobium vesicles or chlorosomes
in green sulfur bacteria
lie close or under the PM (w/c contains enzymes and proteins needed for photosynthesis) but not continuous with it
PHOTOSYNTHETIC MEMBRANES: chromatophore

in purple non-sulfur bacteria
continuous with PM
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PHOTOSYNTHETIC MEMBRANES: photosynthetic lamellae

may be part of the PM or close to it Example: in purple sulfur bacteria gives surface area continuous with PM
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INTERNAL MEMBRANE SYSTEM

The Mesosomes
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MESOSOMES
Invaginations of the plasma membrane in the shape of vesicles, tubules or lamellae Present in both Gram positive and Gram negative bacteria (generally prominent in Gram +) Functions:
Postulated to be involved in cell wall formation during division Play a role in chromosome replication and distribution to daughter cells Involved in secretory processes
STRUCTURES EXTERNAL TO THE CELL WALL

S Layer
Glycocalyx Flagella Axial Filaments
Fimbriae and Pili

THE S LAYER
Paracrystalline surface layer common cell wall layer among Archaea and some Gram positive composed of interlocking proteins / glycoproteins Resistant to penicillin and lysozyme Functions:
protection against ion and pH fluctuations, and osmotic stress, predaceous bacteria selective sieve: allows passage f lowmolecular-weight and excluding large molecules and structures retain proteins near the cell surface
85
THE S LAYER
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GLYCOCALYX
Outside cell wall Usually sticky Extracellular polysaccharide Slime layer and capsule
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WHATS THE DIFFERENCE?

GLYCOCALYX
Network of polysaccharides extending from the surface of bacteria Encompass both capsules and slime layers
CAPSULE
Well-organized layer, not easily washed-off
SLIME LAYER
Diffuse, unorganized material, easily removed
FUNCTIONS OF GLYCOCALYX
protection from phagocytosis
contributes to bacterial virulence (degree to which it can cause disease)
pathogenic-encapsulated protein and polysaccharide-difficult to digest
protection from dessication

water cant escape
adherence to surfaces
in the intestine; can reproduce
source of nutrition
in the mouth; adheres to the teeth if no food available, glycocalyx will be utilized
EXTERNAL FILAMENTOUS APPENDAGES
90
PILI AND FIMBRIAE

FIMBRIAE (fimbria)
Short, fine, hair-like appendages thinner than flagella not involved in motility Visible using EM Composed of helically arranged protein subunits May be used for attachment to surfaces or host
PILI (pilus)
Also called sex pilus, Larger than fimbriae Genetically determined by sex factors or conjugative plasmids Required for bacterial mating
FIMBRIAE AND PILI
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HOW BACTERIA MATE: EXCHANGE OF DNA
93
FLAGELLA AND MOTILITY

FLAGELLA
Threadlike locomotor appendages extending outward from the plasma membrane and cell wall
helical protein filaments responsible for rapid free-swimming motility
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FLAGELLAR ARRANGEMENTS
4 arrangements of flagella are used for identification of bacteria
Monotrichous: 1 flagella at one end
Amphitrichous: both sides of flagella Lopotrichous: path of flagella at one end;
Peritrichous: all over the surface; around;

FLAGELLAR ULTRASTRUCTURE
3 PARTS
FILAMENT
Longest and most obvious, extends from the cell surface to the tip
BASAL BODY
Embedded in the cell
HOOK
Short, curved segment, links filament to its basal body
Flagellin
Protein subunits of filament (MW = 30, 000 - 60, 000)
THE RINGS OF THE BASAL BODY

GRAM POSITIVE GRAM NEGATIVE
L ring P ring S ring M ring
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ROTATION
Hypothesis: a flagellum rotates because of interactions between its S ring and M ring A rod or shaft extends from the hook and ends in the M ring Which can rotate freely in the plasma membrane S ring is attached to the cell wall in Gram + cells and does not rotate A turning motion will result if the two rings interact lecture 3 MICROBIAL CELL BIOLOGY 98
READING ASSIGNMENT
HOW DOES FLAGELLAR SYNTHESIS TAKE PLACE?
99
FLAGELLAR MOVEMENT
flagellar rotation depends on the cells continuous generation of energy (ATP-driven) RUN or SWIM- movement in one direction for a period of time
TUMBLE or TWIDDLE- abrupt or random changes in the direction which interrupts a run or swim
101
CHEMOTAXIS
102
AXIAL FILAMENT AND GLIDING MOTILITY

for gliding or spiral motion; unique to spirochetes
Borrelias Leptospires Treponemes
Endoflagella Anchored at one end of a cell Rotation causes cell to move

104
BACTERIAL ENDOSPORE
a sprecial resistant, dormant structure formed internal to the plasma membrane by certain G+ bacteria highly refractile structure can be seen; no need for staining in brightfield microscopy resistant to freezing temperature, heat, UV, radiation, dessication, chemical disinfectants some may remain viable for hundred or thousands or even millions of years
formed when essential nutrients become depleted

LOCATION AND SIZE
106
PARTS OF AN ENDOSPORE
EXOSPORIUM Thin delicate covering SPORE COAT Lies beneath exosporium; fairly thick Composed of several protein layers Impermeable and resistant to chemicals SPORE CELL WALL Inside the cortex Surrounds the protoplast or core CORE Has the normal cell structure such as ribosomes and nucleoid
CORTEX
Occupies half the spore volume Rests beneath the spore coat Made up of PG that is less cross-linked than vegetative cell lecture 3 MICROBIAL CELL BIOLOGY
107
Attributes of a spore cell

spore cell is dry 10-30% water content dipicolinic acid reacts with calcium ions to form calcium dipicolinate, which is a highly resistant chemical and maintains the dry environment within the cell Produce acid-soluble spore proteins within the cell - protect the cell from desiccation and protect the cells DNA from UV radiation. - serve as the only carbon and energy source upon germination Spores have a tough outer covering made of the protein keratin and are resistant to heat and chemicals. The keratin also resists staining, so extreme measures must be taken to stain the spore.
SPORE RESISTANCE
15% of spore dry weight consists of dipicolinic acid complexed with calcium ions DPA directly involved in spore heat resistance
But recent studies on mutant lacking DPA show resistance
Calcium DPA stabilizes spore nucleic acids Dehydration of the protoplast: important in heat resistance
The cortex may osmotically removed water from protoplast protecting it form heat and radiation damage
FACTORS:
Calcium DPA stabilization of components like DNA, protoplast dehydration, greater stability of cell proteins
THE SPORE FORMERS

Clostridium Bacillus Sporolactobacillus Sporosarcina
Can you name others?
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CYCLES OF SPORULATION AND GERMINATION
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STEPS IN ENDOSPORE FORMATION

Axial filament formation Septum formation Lysis of sporangium Engulfment of forespore Cortex formation Coat synthesis Completion of coat synthesis, increase in refractility and heat resistance
Free spore
Germination and outgrowth
SIGNIFICANCE OF ENDOSPORES
importance of endospores: the bacterium can await for favorable environmental condition good for generation
Tyndallization- endospores can be destroyed by breaking the spore coat through alternate heating and cooling
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THE CYTOPLASMIC MATRIX RIBOSOMES INCLUSIONS
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THE CYTOPLASMIC MATRIX

Substance lying between the plasma membrane and the nucleoid Largely water (about 70%)
Often packed with ribosomes Protoplast: plasma membrane + everything within
Cytoplasmic matrix is a major part of the protoplast
INCLUSION BODIES
Granules of organic or inorganic material that are often clearly visible in a light microscope present in a cytoplasmic matrix
Not bounded by a membrane and lie freely in the cytoplasm Examples: Polyphosphate granules, cyanophycin granules, glycogen granules Enclosed by a single-layered membrane (2-4 nm thick) Examples: PHB granules, some sulfur and glycogen granules, carboxysomes and gas vacuoles
Vary in composition (proteins, lipids, etc)

BACTERIAL INCLUSIONS (INCLUSION BODIES)

Metachromatic granules (volutin) Phosphate reserves Polysaccharide granules Energy reserves Example: Glycogen Lipid inclusions Energy reserves
Sulfur granules
Carboxysomes Gas vacuoles
Energy reserves
Ribulose 1,5-diphosphate carboxylase for CO2 fixation Protein covered cylinders
Magnetosomes
Iron oxide (destroys H2O2)

METACHROMATIC GRANULES
Example: volutin
Corynebacterium diptheriae
turns red if you use blue dye

represent reserves of inorganic phosphate used in ATP synthesis found also in algae, fungi, and protozoans
LIPID INCLUSIONS
common lipid storage is PHB (poly- hydroxybutyric acid)
detected by Sudan dyes ex. Mycobacterium, Bacillus, Azotobacter, Spirillum
SULFUR INCLUSIONS
energy reserve for sulfur bacteria Example: Thiobacillus
sulfur bacteria, derive energy by oxidizing sulfur and sulfur-containing compound
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CARBOXYSOMES
contain the enzyme RDP carboxylase
used by bacteria in CO2 fixation during photosynthesis
Example: Nitrifying bacteria, cyanobacteria, Thiobacillus
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GAS VACUOLES
Gas vesicles
hollow cavities found in aquatic prokaryotres, cyanobacteria, anoxygenic photosynthesizers and halobacteria maintain buoyancy of aquatic cells
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MAGNETOSOMES
inclusions of iron oxides (Fe3O4) found in some Gbacteria such as Aquaspirillum magnetotacticum that act like magnets
used by bacteria to orient themselves in the earths magnetic field bacteria use the magnetosome chain to determine northward and downward directions, and swim down to nutrient rich sediments or locate the optimum depth in freshwater and marine habitats
MAGNETOSOMES
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RIBOSOMES
Loosely attached to the plasma membrane Made up of protein and ribonucleic acid (RNA) Site of protein synthesis 2 subunits as 50S and 30S (S = Svedberg unit; unit of sedimentation) 70S = 50S + 30S
Svedberg units is not directly proportional to molecular weights
THE 16S COMPONENT USED BY CARL WOESE

ribosomes (proteins + rRNA) of bacteria
70S proteins RNA = 30S 21 16S + 50S 34 5S 23S
129
THE BACTERIAL RIBOSOME
130
THE NUCLEOID
Nuclear region
3% of cell weight 10% of cell volume
no histones present
chromosome are attached directly to the PM contains small, circular DNA molecules called PLASMIDS, believed to be associated with membrane proteins
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contains bacterial chromosome which is a single, long, circularly arranged thread of DNA
THE NUCLEOID
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PLASMIDS
In addition to their chromosomes
contain 5 to 100 genes responsible for activities such as:
antibody resistance tolerance to toxic metals synthesis of enzymes production of toxins gene transfer
DEFINITION OF TERMS
Protoplasts
A bacterial or fungal cell with its cell wall completely removed; spherical; osmotically sensitive
Spheroplasts
A relatively spherical cell formed by weakening or partial removal of the rigid cell wall component (e.g. penicillin treatment); osmotically sensitive
L-forms
Pleiomorphic bacterial cells formed by the complete or partial loss of their cell walls; cell wall loss may be reversible or permanent
THE EUKARYOTES
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Organelles
Membrane-bound:
Nucleus ER Golgi complex Lysosome Vacuole Mitochondrion Chloroplast Peroxisome Contains chromosomes Transport network Membrane formation and secretion Digestive enzymes Brings food into cells and provides support Cellular respiration Photosynthesis Oxidation of fatty acids; destroys H2O2
Ribosomes
80S
Membrane-bound Free Attached to ER In cytoplasm
70S
In chloroplasts and mitochondria
Golgi Complex
Figure 4.26
Lysosomes
Figure 4.22b
Vacuoles
Figure 4.22b
Mitochondrion
Figure 4.27
Chloroplast
Figure 4.28
INCLUSIONS
polysaccharide granules
consists of glycogen and starch glycogen granules turn reddish brown and starch appear blue with iodine
Magnetosomes
also present in heads of birds, tuna, dolphins, and green turtles two kinds: magnetite: Fe3O4 for direction, i.e. Northern hemisphere or Southern Hemisphere
gregite: Fe3S4
READING ASSIGNMENT
Difference between prokaryotes and Eukaryotes in terms of: 1. Use of membranes 2. General differences

Lecture 3

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Microbial Cell Biology

Lecture 3 Lani Manahan-Suyom

HOW DO WE OBSERVE MICROBIAL CELLS?

A microscope with a resolving power of 0.2 m (200 nm)

THE USE OF OIL IMMERSION

lecture 3 MICROBIAL CELL BIOLOGY

lecture 3 MICROBIAL CELL BIOLOGY

PHASE CONTRAST MICROSCOPY

lecture 3 MICROBIAL CELL BIOLOGY

DIFFERENTIAL INTERFERENCE MICROSCOPY

lecture 3 MICROBIAL CELL BIOLOGY

Cells may be stained with fluorescent dyes (fluorochromes)

lecture 3 MICROBIAL CELL BIOLOGY

lecture 3 MICROBIAL CELL BIOLOGY

TRANSMISSION ELECTRON MICROSCOPY (TEM)

10,000-100,000; resolution 2.5 nm

lecture 3 MICROBIAL CELL BIOLOGY

SCANNING ELECTRON MICROSCOPY

lecture 3 MICROBIAL CELL BIOLOGY

SCANNING TUNNELING MICROSCOPY

lecture 3 MICROBIAL CELL BIOLOGY

ATOMIC FORCE MICROSCOPY

lecture 3 MICROBIAL CELL BIOLOGY

Looking into cell details

Cell Morphology (shape)

CELL DIVISION AND SHAPES: COCCI

lecture 3 MICROBIAL CELL BIOLOGY

CELL DIVISION AND SHAPES: COCCI

lecture 3 MICROBIAL CELL BIOLOGY

CELL DIVISION AND SHAPES: COCCI

lecture 3 MICROBIAL CELL BIOLOGY

CELL DIVISION AND SHAPES: SPIRALS

lecture 3 MICROBIAL CELL BIOLOGY

OTHER SHAPES OF BACTERIA

lecture 3 MICROBIAL CELL BIOLOGY

lecture 3 MICROBIAL CELL BIOLOGY

lecture 3 MICROBIAL CELL BIOLOGY

CELL DIVISION AND SHAPES: BACILLI

lecture 3 MICROBIAL CELL BIOLOGY

Importance of cell size?

PROKARYOTIC CELL ORGANIZATION

lecture 3 MICROBIAL CELL BIOLOGY

TYPICAL BACTERIAL CELL

lecture 3 MICROBIAL CELL BIOLOGY

FUNCTIONS OF PROKARYOTIC STRUCTURES

lecture 3 MICROBIAL CELL BIOLOGY

FUNCTIONS OF PROKARYOTIC STRUCTURES

Fimbriae and Pili

FUNCTIONS OF PROKARYOTIC STRUCTURES

Localization of Genetic Material (DNA)

THE PROKARYOTIC CELL WALL

lecture 3 MICROBIAL CELL BIOLOGY

IMPORTANCE OF CELL WALLS FOR BACTERIA

CHRISTIAN GRAM AND HIS STAIN

Divided the microbial world into 2: positive and negative

lecture 3 MICROBIAL CELL BIOLOGY

Profile of the bacterial cell envelope

DIFFERENTIATING GRAM POSITIVE AND NEGATIVE WALLS

lecture 3 MICROBIAL CELL BIOLOGY

Chemical nature of bacterial cell walls

Chemical nature of bacterial cell walls

Chemical nature of bacterial cell walls

Chemical nature of bacterial cell walls

Lysozyme breaks this glycoside bond between M and G