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MICROBES. Microscopic life forms that cannot be seen by the unaided eye Magnifying device Microscope . “illumination” Light (visible) Fluorescent U-V Electron. eyepieces Monocular Binocular. specimen background Bright field Dark field. MICROSCOPES. Resolving Power N.A. affects
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MICROBES • Microscopic life forms that cannot be seen by the unaided eye • Magnifying device • Microscope
“illumination” Light (visible) Fluorescent U-V Electron eyepieces Monocular Binocular • specimen background • Bright field • Dark field MICROSCOPES • Resolving Power • N.A. affects • Wavelength affects MICROSCOPES
Types of Scopes MICROSCOPES
Types of Scopes MICROSCOPES
PARTS & TERMS • Lenses (ocular, objective) • Total Magnification • ocular mag. X objective mag. • Condenser • Diaphram MICROSCOPES
Tube Length Adjustment Ring Eyepiece Binocular Body Clamp Screw Nosepiece Microscope Limb/Arm Objective Tension Control Knob Mechanical Stage Coarse Focus Knob Substage Condenser with Iris Diaphragm Fine Focus Knob Power Switch & Brightness Control Knob Microscope Base Illuminator Condenser Mechanical Stage Control Knob Substage Focus Control Knob Field Iris
PARTS & TERMS • Numerical Aperture (N.A.) • optical characteristic of a lens • N.A. = i sinθ N.A. increases with magnification MICROSCOPES
(l innm) R.P. = wavelength of illumination (2)N.A. PARTS & TERMS • Resolving Power (R.P.) • size of the smallest discernable detail • minimum distance between objects so that they are able to be distinguished as separate [ RP of human eye ~ 0.2mm = ______m ] MICROSCOPES
l = 650nm, NA = 1.25 R.P. = l/2NA = _____nm = _____ m l = 450nm, NA = 1.25 R.P. = l/2NA = _____nm = _____ m Resolving Power (R.P.) l= 650nm, NA = 0.25 R.P. = l/2NA = _____nm = _____ m MICROSCOPES
Resolving Power (R.P.) • R.P. = l/2NA Larger or Smaller R.P. = BETTER ? • R.P. improves as N.A. increases or decreases? • R.P. improves as l increases or decreases? MICROSCOPES
Types of Scopes • Compound Light • Bright or Dark field • Use blue light or blue filter for shorter l • R.P. ~ 0.2 m MICROSCOPES
Types of Scopes • Fluorescent • Dark field compound light microscope • Uses UV for side illumination of specimen, fluorescent parts or dyes give off visible light that is viewed • R.P. ~ 0.2 m MICROSCOPES
Types of Scopes • UltraViolet (UV) • Uses UV as illumination (shorter l) • Image recorded then viewed… still or video camera image(should NOT look directly at UV!) • Special lens material (quartz),glass absorbs UV • R.P. ~ 0.15 m MICROSCOPES
Types of Scopes • Electron Microscopes • Use electron beam for illumination, magnets for “lenses”, video to view • Transmission Electron Microscope (TEM) • beam passes thru, view internal structure • Scanning Electron Microscope (SEM) • beam reflects, see external structure, texture MICROSCOPES
Transmission Electron Microscope (TEM) MICROSCOPES
Types of Scopes • Transmission Electron Microscope (TEM) • Special specimen preparation (vacuum) • Working magnification: • whole bacteria….. 8,000x - 10,000x • thin section, viruses…. 30,000x - 40,000x • Maximum magnification ~100,000x • R.P. ~0.001 m = 1 nm MICROSCOPES
rickettsia herpes simplex
Scanning Electron Microscope (SEM) MICROSCOPES
Types of Scopes • Scanning Electron Microscope (SEM) • Special specimen preparation(vacuum & conductor) • Working magnification: • 14,000x - 50,000x • Maximum magnification ~130,000x • R.P. ~0.01 m = 10 nm MICROSCOPES
Q fever (rickettsia) Image enhancement
MICROBES • Microscopic life forms that cannot be seen by the unaided eye • May be plant or animal • May be prokaryotic or eukaryotic
Importance of Microbes • Pathogenic • Help cure / prevent disease • Break down organic matter to inorganic forms (decompose, recycle) • Autotrophs (inorganics) • Heterotrophs (organics) • Decomposers (inorganics, external digestion!)
Importance of Microbes • Spoil foods • Make / flavor foods • Petroleum & Coal: unrecycled remnants • Decay wooden structures • Corrode iron & steel, damage concrete • Food source • Allow fouling community establishment
Cell Membrane Cytoplasm Organelles/ Vacuoles Cell Nucleus Chloroplasts Cell Wall Classification of Microbes Plant vs Animal cell
Nuclear Membrane Classification of Microbes Prokaryotic vs Eukaryotic
Classification of Microbes Domains and Kingdoms Prokaryotic • Domain • Archaea • Domain • Bacteria • Eukaryotic • Domain Eukarya • Protista • Fungi • Plantae • Animalia
Archaea[Prokaryotic] • Crenarcharota • Euryarchaeota • Korarchaeota • Subtypes: • Methanogens • Halophiles • Thermophiles • Psychrophiles
Archaea[Prokaryotic] • Methanococcus jannaschii • 1st archaea to be genetically sequenced
Bacteria[Prokaryotic] • Cyanobacteria • Blue-green algae
Bacteria Morphology (shapes) • Coccus (cocci) • spheres • Bacillus (bacilli) • rods • Spirillus (spirilli) • spiral, corkscrew (spirochetes)
Gonorrhea pneumonia Staph infections Bacteria Morphology (shapes) Coccus (cocci) • diplococcus • streptococcus • staphylococcus
E.coli cholera meningitis Legionnaire’s disease Bacteria Morphology (shapes) Bacillus (bacilli)
peptic ulcers salmonella whooping cough tuberculosis Bacteria Morphology (shapes) Bacillus (bacilli)
Bacteria Morphology (shapes) Spirillus (spirilli)
Protista [Eukaryotic] • Algae, Protozoa, Slime Molds & Water Molds
Fungi [Eukaryotic] • Molds • mycelia = ‘body’ • hyphae = threads
Fungi [Eukaryotic] • Molds
Fungi [Eukaryotic] • Molds • Yeast
Fungi [Eukaryotic] • Molds • Yeast • Mushrooms
Lab Procedures • Bunsen burners