Outline of Todays Lecture

1. Microscopy and Specimen Preparation

2. Prokaryotic Cell Structure and

Function (Cell Envelope)
 The Study of Microbial Structure:
Microscopy and Specimen Preparation
 The Light Microscope
Many types
bright-field microscope
dark-field microscope
phase-contrast microscope
fluorescence microscopes
 Preparation and Staining of
Specimens

Increases visibility of specimen

Accentuates specific morphological features
Preserves specimens
 Fixation

process by which organism is killed and firmly

attached to microscope slide
heat fixing
preserves overall morphology but not internal structures
 Dyes and Simple Staining

simple staining
a single staining agent is used
basic dyes are frequently used
dyes with positive charges
e.g., crystal violet
methylene blue
 Differential Staining

divides microorganisms into groups

based on their staining properties
e.g., Gram stain
e.g., acid-fast stain (mainly Mycobacteria)
 primary
stain

mordant

decolorization

counterstain
positive
negative
 Gram staining
most widely used differential staining
procedure (name after the Danish
Scientist Hans Christian Gram
1853-1938)

divides Bacteria into two groups based

on differences in cell wall structure
Clinical samples
Prokaryotic Cell Structure
and Function
 Size of Bacteria
Average bacteria 0.5 - 2.0 um in diam.
 Shapes of Bacteria

Coccus
Chain = Streptoccus
Cluster = Staphylococcus
Coccobacillus
Vibrio = curved
Bacillus
Chain = Streptobacillus
Spirillum
Spirochete
 Bacterial Structures

Flagella
Pili
Capsule
Plasma Membrane
Cytoplasm
Cell Wall
Lipopolysaccharides
Teichoic Acids
Inclusions
Spores
Bacterial Cell Envelope Structure
Gram Positive Bacteria Cell Envelope
Gram Negative Bacteria Cell Envelope
 Prokaryotic Cell Membranes

membranes are an absolute requirement for all

living organisms
plasma membrane encompasses the cytoplasm
some prokaryotes also have internal membrane
systems
Phospholipids
Major components of the
cell membrane
Consists of two fatty
acids and the third
hydroxyl group of
glycerol is joined to a
phosphate group
Amphipathic
- hydrocarbons tail
- phosphate head
The asymmetry of most membrane
lipids

polar ends
interact with water
hydrophilic
nonpolar ends
insoluble in water
hydrophobic
 The plasma membrane

separation of cell from its environment

selectively permeable barrier
some molecules are allowed to pass into or
out of the cell
transport systems aid in movement of
molecules
 Periplasmic space

gap between plasma membrane and

cell wall (gram-positive bacteria)
or between plasma membrane and
outer membrane (gram-negative
bacteria)
periplasm
substance that occupies periplasmic space
Gram Negative Bacteria Cell Envelope
 Periplasmic enzymes

found in periplasm of gram-negative

and gram-positive bacteria
 Periplasmic enzymes

Some of their functions

nutrient acquisition
electron transport
peptidoglycan synthesis
modification of toxic compounds
 Exoenzymes
secreted by bacteria to the outside of the cell
perform many of the same functions that
periplasmic enzymes do for bacteria
 The Prokaryotic Cell Wall

rigid structure that lies just

outside the plasma
membrane
the determination of cell
shape
prevent bursting from
osmotic pressure (lysis)
bacterial cytoskeleton?
Bacterial cytoskeleton
 Cell walls of Bacteria

Bacteria are divided into two major

groups based on the response to Gram-
stain procedure.
gram-positive bacteria stain purple
gram-negative bacteria stain pink
staining reaction due to cell wall
structure
 The Mechanism of Gram Staining

involve constriction of the thick peptidoglycan

layer of gram-positive cells
constriction prevents loss of crystal violet
during decolorization step

thinner peptidoglycan layer of gram-negative

bacteria does not prevent loss of crystal violet
 Cell Wall
Important component of both gram-positive and
gram-negative bacteria

Unique to bacteria

Peptido-glycan Polymer (amino acids + sugars)

Sugars; NAG & NAM

N-acetylglucosamine
N-acetymuramic acid

Amino acids cross link NAG & NAM

E. coli

S. aureus

DAP(diaminopimelic acid)
A Very Complex Net
Bacterial Cell Envelope Structure