Microbial Cell Structure and Function Flashcards
uses visible light to illuminate cells
Compound light microscope
Many different types of light microscopy:
Bright-field
Phase-contrast
Dark-field
Fluorescence
the ability to make an object larger
Magnification
Maximum magnification is
~2,000✕
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
Resolution
Limit of resolution for light microscope is about
0.2 μm
Specimens are visualized because of differences in
contrast (density) between specimen and surroundings
Bright-field scope
improves contrast
Staining
are organic compounds that bind to specific cellular materials
Dyes
Examples of common stains are
methylene blue, safranin, and crystal violet
separate bacteria into groups
Differential stains
Bacteria can be divided into two major groups:
gram-positive and gram-negative
appear purple
Gram-positive bacteria
appear red after staining
gram-negative bacteria
Phase-contrast microscopy was invented in ______________ by ________________________
1936 by Frits Zernike
Improves the contrast of a sample without the use of a stain
Phase-contrast microscopy
Phase ring amplifies differences in the refractive index of cell and surroundings
Phase-contrast microscopy
Allows for the visualization of live samples
Phase-contrast microscopy
Light reaches the specimen from the sides
Dark-field microscopy
Light reaching the lens has been scattered by specimen
Dark-field microscopy
Image appears light on a dark background
Dark-field microscopy
Excellent for observing motility
Dark-field microscopy
Used to visualize specimens that fluoresce
Fluorescence microscopy
Specific on staining DNA
DAPI
use electrons instead of photons to image cells and structures
Electron microscopes
Two types of electron microscopes:
Transmission electron microscopes (TEM)
Scanning electron microscopes (SEM)
- Electromagnets function as lenses
- System operates in a vacuum
- High magnification and resolution (0.2 nm)
- Enables visualization of structures at the molecular level
- Specimen must be very thin (20–60 nm) and be stained
(osmic acid,permanganate, uranium lanthium or lead
salts)
Transmission electron microscopy (TEM)
- 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 - Even very large specimens can be observed
- Magnification range of 15✕–100,000✕
Scanning electron microscopy (SEM)
cell shape
Morphology
Major cell morphologies
- Coccus (pl. cocci)
- Rod
- Spirillum
spherical or ovoid
Coccus (pl. cocci)
cylindrical shape
Rod
spiral shape
Spirillum
Cells with unusual shapes
Spirochetes, appendaged bacteria, and filamentous bacteria
Size range for prokaryotes
0.2 µm to >700 µm in diameter
Examples of very large prokaryotes
- Epulopiscium fishelsoni
- Thiomargarita namibiensis
Size range for eukaryotic cells:
10 to >200 µm in diameter
Thin structure that surrounds the cell
Cytoplasmic membrane
Vital barrier that separates cytoplasm from environment
Cytoplasmic membrane
Highly selective permeable barrier; enables concentration of specific metabolites and excretion of waste products
Cytoplasmic membrane
Firmly embedded in the membrane
Integral membrane proteins
One portion anchored in the membrane
Peripheral membrane proteins
Ether linkages in phospholipids of Archaea
Archaeal membranes
Major lipids are
glycerol diethers and tetraethers
Holds transport proteins in place
Protein anchor
Generation of proton motive force
Energy conservation
What are used for magnification?
Objective lens and ocular lens
Total magnification =
objective magnification ✕ ocular magnification
Differential stains:
the Gram stain
Resulting image is dark cells on a light background
Phase-contrast microscopy
Dark-field microscopy is excellent for observing __________________
motility
Emit light of one color when illuminated with another color
of light
fluoresce
Cells fluoresce naturally..example
Chlorophyll
Widely used in microbial ecology for enumerating
bacteria in natural samples
Fluorescence microscopy
Can stain both live (less efficient) and fixed cells
DAPI
Excited by an ultraviolet light (358nm) emit blue
color (461nm).
DAPI
Enables visualization of structures at the molecular level
Transmission electron microscopy (TEM)
Surface area:
increased 4x
Volume:
increased 8x
Composition of membranes
* General structure is ___________________________
phospholipid bilayer
how wide is cytoplasmic membrane
8–10 nm wide
Cytoplasmic membrane is stabilized by
hydrogen bonds and hydrophobic interactions
help stabilize membrane by forming ionic
bonds with negative charges on the phospholipids
Mg2+ and Ca2+
Ether linkages in phospholipids of Archaea
Archaeal membranes
Bacteria and Eukarya that have ester linkages in
phospholipids
Archaeal membranes
Archaeal lipids lack ________________; have ________________ instead
fatty acids; isoprenes
contains 4 5 carbon cyclopentyl and 1 6 carbon cylohexyl
Crenarcheol
Functions of the cytoplasmic membrane
- Permeability barrier
- Protein anchor
- Energy conservation
- Show saturation effect – capacity of the carrier proteins
- Highly specific – accommodate specific or closely (ex.
Amino acid) related molecules - Highly regulated
Carrier-mediated transport systems
Three major classes of transport systems in prokaryotes
- Simple transport
- Group translocation
- ABC system
Driven by the energy in the proton motive force
Simple transport
Chemical modification of the transported substance driven by phosphoenolpyruvate
Group translocation
Periplasmic binding proteins are involved and energy comes from ATP
ABC transporter
Three transport events are possible:
uniport, symport, and antiport
transport in one direction across the membrane
uniport