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8/9/2019 Microscopy, Cell Structure and 5 Groups of Microbes
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Chapter
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Connects theeyepiece to the
objective lenses
d to reflect light
anexternal light
urceup throug
10X or 15X power.
Supports the tubeandc
to thebase
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nsist of 4X, 10X, 40X and 100X powers.
itha 10X (most common) eyepiece lens,magnifications40X (4X times 10X), 100X , 400X and 1000X.
O - shortest lens- longest one with the greatest power.
nsesarecolorcodedand ifbuilt to DIN standardsarerchangeablebetweenmicroscopes.
hehighpowerobjective lensesare retractable(i.e. 40Xey hit aslide, theendof the lens will push in(spring lo
reby protecting the lensand theslide.
0
0X powers. epiece lens,magnifications
400X and 1000X.
atest power.
built to DIN standardsarescopes.
esare retractable(i.e. 40Xlens will push in(spring lo
theslide.
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Rack Stop:
- anadjustment that determineshow close
theobjective lenscan get to theslide.
- only need toadjust if youareusing very
thinslidesand you weren't able to focusonthespecimenat highpower.
etermineshow close
get to theslide.
youareusing very
ren't able to focusonpower.
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n enser ens:purposeto focus the light onto the specimen.
most useful at thehighest powers(400X andabov
phragm or Iris:
most microscopes havea rotating diskunder thtage.
thisdiaphragmhasdifferent sizedholesand isuso vary the intensity and size of the cone of lighhat is projected upward into the slide.
ht onto the specimen.
t powers(400X andabov
a rotating diskunder th
ent sizedholesand isussize of the cone of lighinto the slide.
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Bright-fieldmicroscopy
Most common
Visible light
Maxmagnification is
about 1,000xPhasecontrast
Usehigh-contrast
objectives
Bright-fieldmicroscopy
Most common
Visible light
Maxmagnification is
about 1,000xPhasecontrast
Usehigh-contrast
objectives
D k fi ld
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Dark-fieldx side illuminationx darkbackground, light specimen
Fluorescencemicroscopex Requiresstaining ofspecimen with
fluorescent dyex Highermagnification thanbright field
Confocal scanning lasermicroscopex Computercontrolledx Laser light sourcex Fluorescentx Cansequentially imagemultipleplanes
ofasampleandprovide D images
ith
field
planeses
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transmission(TEM)
scanning (SEM)
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` Stains All stainsmust haveoneormoredyes
x Basicdyeshaveapositivecharge
x Acidicdyeshaveanegativecharge
Most stainsareperformedon glassslides
` Simplestains
Usually useabasicdye that providescolorandcontrast toacolorlessmicrobe
` Stains All stainsmust haveoneormoredyes
x Basicdyeshaveapositivecharge
x Acidicdyeshaveanegativecharge
Most stainsareperformedon glassslides
` Simplestains
Usually useabasicdye that providescolorandcontrast toacolorlessmicrobe
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m cro es Gramstain separatesbacteria intoGrampositi
(purple) ornegative( red)
x Primary stain iscrystal violet (purple)x Secondary stain issafranin (reddye)
sbacteria intoGrampositi
red)
al violet (purple) franin (reddye)
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ono rea y s a n w ram ss a nThebacterial cellsare flooded withcarbol fuchsin (red) anheat isapplied
Thecellsare rinsed withacidalcohol, which removescarfuchsin fromcells that arenot acid-fast
Methyleneblue isusedasacounterstain
ram ss a nd withcarbol fuchsin (red) an
alcohol, which removescart acid-fast
counterstain
s r c res
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s ruc ures Malachite green isused tostainendospores
India ink isused tostaincapsularbacteria
d tostainendospores
incapsularbacteria
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` Fluorescent dyes
DNA-binding dyes
x acridine orangex ethidium bromide
Immunofluorescence reliesuponafluorescently-labeledantibody specific toa
cellularcomponent
liesuponantibody specific toa
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a. Bacteria
b. fungi: yeasts
and molds
c. Viruses
d. protozoa
e. algae
a. Bacteria
b. fungi: yeasts
and molds
c. Viruses
d. protozoa
e. algae
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ria are typically
llular,
scopic,
ryoticorganismseproduceby
fission
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` Note gram-positive(purple) cocciclusters.
m-positive(purple) cocci.
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Yeasts are typicallyunicellular,microscopic,eukaryotic fungi thatreproduceasexually bybudding
` Molds are typicallyfilamentous,eukarfungi that reproducproducing asexualreproductivespore
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Saccharomyces cerevisiae
Note budding yeast (arrows).
Wine yeast withbudandbudscars(Sacchr
spp.).
Magnification:-- x2,270--(Basedonan image
inch in thenarrow dimension)
Wine yeast withbudandbudscars(Sacchr
spp.).
Magnification:-- x2,270--(Basedonan image
inch in thenarrow dimension)
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ly submicroscopic,ar infectiousparticlesnonly replicatea living host cell.
st majority of virusesseither DNAor RNA
t both.
Transmission Electron Microgra
Adenovirus
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pically
llular,
scopic,
ryotic
isms that lack
l wallPhotomicrograph ofAmoeb
proteus
Note the numerous pseudopodi
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pically eukaryotic
organisms that
out photosynthesis
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` ree oma n ys em y oese e am
y oese e a
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` ree oma n ys em y oese e a isanevolutionary model ofclassificationbased
x differences in1. the sequences of nucleotides in the cell's ribos
RNAs (rRNA)
2. the cell's membrane lipid structure
3. its sensitivity to antibiotics.
m y oese e a del ofclassificationbased
f nucleotides in the cell's ribos
ane lipid structure
antibiotics.
`
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different cell types,each representing adomain.
` The threedomainsare
theArchaea (archaebacteria), theBacteria (eubacteria),and theEukarya (eukaryotes).
x 4 kingdoms:x Protistsx FungixAnimaliax Plantae
h representing adomain.
bacteria),ria),andtes).
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Archaea possess the
llowing characteristics:
rokaryotic cells
havemembranescomposedofbranched hydrocarbonchains attached toglycerol by ether linkages
Archaea possess the
llowing characteristics:
rokaryotic cells
havemembranescomposedofbranched hydrocarbonchains attached toglycerol by ether linkages
`
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.
` Not sensitive tosomeantibiotics that affect theBabut aresensitive tosomeantibiotics that affect theEukarya.
` Contain rRNA that is unique to theArchaea as iby thepresencemolecular regionsdistinctly differthe rRNAofBacteria andEukarya.
` Live inextremeenvironmentsand includemethan
extremehalophiles,andhyperthermophiles.
.
antibiotics that affect theBa omeantibiotics that affect the
unique to theArchaea as i cular regionsdistinctly differ
andEukarya.
onmentsand includemethan
ndhyperthermophiles.
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cteria possess the following characteristics:teria areprokaryotic cells.
theEukarya, they havemembranescomposedofunbranchedacid chains attached to glycerol by ester linkages
wallscontain peptidoglycan.
itive to traditional antibacterial antibioticsbut are resistant tot antibiotics that affect Eukarya.
ain rRNA that is unique to the Bacteria as indicatedby the
encemolecular regionsdistinctly different from the rRNA ofaea andEukarya.
mplesaremycoplasmas,cyanobacteria,Gram-positivebacteriaGram-negativebacteria.
cteria possess the following characteristics:teria areprokaryotic cells.
theEukarya, they havemembranescomposedofunbranchedacid chains attached to glycerol by ester linkages
wallscontain peptidoglycan.
itive to traditional antibacterial antibioticsbut are resistant tot antibiotics that affect Eukarya.
ain rRNA that is unique to the Bacteria as indicatedby the
encemolecular regionsdistinctly different from the rRNA ofaea andEukarya.
mplesaremycoplasmas,cyanobacteria,Gram-positivebacteriaGram-negativebacteria.
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` possess the following characteristics:
` Eukarya haveeukaryotic cells.
` Like theBacteria, they havemembranescomposedofunbrafatty acid chains attached to glycerol by ester linkages
` cell wall contains no peptidoglycan.` resistant to traditional antibacterial antibioticsbut aresensitiv
antibiotics that affect eukaryoticcells.
` contain rRNA that is unique to the Eukarya as indicatedbypresencemolecular regionsdistinctly different from the rRNAArchaea andBacteria.
` possess the following characteristics:
` Eukarya haveeukaryotic cells.
` Like theBacteria, they havemembranescomposedofunbrafatty acid chains attached to glycerol by ester linkages
` cell wall contains no peptidoglycan.` resistant to traditional antibacterial antibioticsbut aresensitiv
antibiotics that affect eukaryoticcells.
` contain rRNA that is unique to the Eukarya as indicatedbypresencemolecular regionsdistinctly different from the rRNAArchaea andBacteria.
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,otic organisms
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,oticorganisms.
ples includesslimemolds,oids,algae,andprotozoans.
ingdomllularormulticellularorganismskaryoticcell types.
avecell wallsbut arenot
ed into tissues.t carry out photosynthesisandnutrients throughabsorption.
ples includesac fungi,clubeasts,andmolds.
cells.cells.
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-cellsareorganized into tissuesandhavewalls.
- obtainnutrientsby photosynthesisandabsorption.
-Examples includemosses, ferns,conifers,flowering plants.
d. Animalia Kingdom- multicellularorganismscomposedofeuk
cells.
- cellsareorganized into tissuesand lackwalls.
- donot carry out photosynthesisandobtainutrientsprimarily by ingestion.
-Examples includesponges, worms, insectvertebrates.
-cellsareorganized into tissuesandhavewalls.
- obtainnutrientsby photosynthesisandabsorption.
-Examples includemosses, ferns,conifers,flowering plants.
nimalia Kingdom- multicellularorganismscomposedofeuk
cells.
- cellsareorganized into tissuesand lackwalls.
- donot carry out photosynthesisandobtainutrientsprimarily by ingestion.
-Examples includesponges, worms, insectvertebrates.
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a. prokaryotic.
b. single-celled,microscopicorganisms
Exceptions - visible to thenakedeye
- Epulopiscium fishelsoni - abacillus, 80 (diameterand 200-600 m long
- Thiomargarita namibiensis - asperical bbetween 100 and 750 m indiameter.
a. prokaryotic.
b. single-celled,microscopicorganisms
Exceptions - visible to thenakedeye
- Epulopiscium fishelsoni - abacillus, 80 (diameterand 200-600 m long
- Thiomargarita namibiensis - asperical bbetween 100 and 750 m indiameter.
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generally much
mallerthanukaryoticcells.
very complexespite theirsmallize. Bacteria on a Human Epithelial
Cell from the Mouth
The bacteria are the small dark purpl
dots and dashes on the light blue cell
The oval purple mass in the center is
nucleus of the epithelial cell.
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` three basic shapes:
Coccus
rodorbacillus
spiral
:
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spherical oroval
acteria
having oneofseveral
istinct arrangements
asedon theirplanesof
ivision.
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lococcus -cocci arranged in
rs
lococcus -cocci arranged in
rs
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tococcus:cocci arranged inchainstococcus:cocci arranged inchains
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d:cocci arrangedaresof 4
ement:
Stain
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lococcus:cocci arranged in irregular,often-likeclusterslococcus:cocci arranged in irregular,often
-likeclusters
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` Electron Micrograph ofMethicillin-Resistant Staphyaureus (MRSA)
h ofMethicillin-Resistant Staphy
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hapedbacteria.
ide inoneplanecing abacillus,tobacillus,orobacillus
gement.
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micrographofabacillus Scanning Electron MicrogrPseudomonas aerugino
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` Escherichia coliO157H7 isastrcoliproducesashiga-like toxin tepithelial cellsof the large intesticausing hemorrhagiccolitis,abl
diarrhea.
` In rarecases, theshiga-toxinenbloodand iscarried to thekidneusually inchildren, it damages vcellsandcauseshemolyticuremsyndrome. Notedividing bacilli.
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c. acoccobacillus:
oval andsimilar toacoc
cillus:
imilar toacoc
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helical orcorkscrew-shapedbacterium.
piralscome inoneof three forms:avibrioaspirillum
aspirochete.
dbacterium.
orms:
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appearsasacurvedbacillus
s).
Vibrio cholerae - Gram-negative,
facultatively anaerobic,curved(vib
shaped), rodprokaryote;causesAcholera.
Vibrio cholerae - Gram-negative,
facultatively anaerobic,curved(vib
shaped), rodprokaryote;causesAcholera.
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rocheteBorrelia (arrows) ina
mear.
Scanning Electron Micrograph ofL
interrogans
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` a typical bacteriumusually consistsof:
` acytoplasmicmembranesurroundedby apeptidoglcell wall andmaybeanoutermembrane;
` a fluidcytoplasmcontaining anuclear region(nuclenumerous ribosomes;and
` often variousexternal structuressuchasa glycocal
flagella,andpili.
` a typical bacteriumusually consistsof:
` acytoplasmicmembranesurroundedby apeptidoglcell wall andmaybeanoutermembrane;
` a fluidcytoplasmcontaining anuclear region(nuclenumerous ribosomes;and
` often variousexternal structuressuchasa glycocal
flagella,andpili.
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` Cytoplasmic Membrane
` Nucleus
` Cytopl
asm
` Cytoplasmic Membrane
` Nucleus
` Cytopl
asm
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` http://www.microbelibrary.org/microbelib
es/ccImages/Articleimages/Spencer/sp
cellwall.html
` http://www.microbelibrary.org/microbelib
es/ccImages/Articleimages/Spencer/sp
cellwall.html
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Shapes
Spherical - coccus (pl.,cocci)
Cylindrical - rod
x very short - coccobacillus
x curved - vibrio
x short spiral - spirillum
x long spiral - spirochete
Shapes
Spherical - coccus (pl.,cocci)
Cylindrical - rod
x very short - coccobacillus
x curved - vibrio
x short spiral - spirillum
x long spiral - spirochete
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