Download - Microbiology Review 1
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Define microscopy
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Microscopy
The usage of microscopes to viewmicroorganisms
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Common units of
measurement
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The Metric System
1m = 1m
1000mm = 1m
10^-6mm = 1 micron
10^-9mm = 1 nanometer
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Define magnification
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Magnification
allows microscopic specimens to becomevisible to the nake e!e
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Define resolution
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Resolution
the abilit! to resolve two ob"ects asseparate# or istinguish ob"ects that are
close together$
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%hat is the resolution
istance epenent on&
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Resolution
epens on the wavelength of
electromagnetic ration# an the
numerical aperture of the lens 'i$e$ the
abilit! of a lens to gather light(
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Define contrast
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Contrast
the ifferences in intensit! between thespecimen an its backgroun
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) wa!s to enhance contrast
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Contrast
1$ staining techni*ues)$ in-phase wavelengths
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%hat are the limits of light
microscop!&
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The limits of light microscopy
1$ ma+ )000+ magnification)$ resolution is limite own to 0$) microns,
an!thing less than that = no$
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+plain 'a( light refraction# an'b( why we use an immersion oil
when using light microscopes
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The refraction of light
.ight passing through a lens refracts# orbens# because light ra!s slow own asthe! enter the glass$ /owever# it oesntcapture some light ra!s which is wh! we
use immersion oils$
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Immersion oil
taking into account that light passes through
immersion oil at the same spee as it
oes with glass# immersion oil is use as a
brige between the slie so light travels
at a uniform spee
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Bright-field microscope
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Bright field microscope
2 illuminate backgroun
2 stain cells to increase contrast
2 cheap# eas!-to-use
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Dark-field microscope
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Dark-field microscope
2 light that is reall! reflecte is magnifie2 no stain neee
2 specimen appears light against a arkbackgroun
2 goo for cells that move
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Phase contrast microscope
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Phase microscope
2 in-phase lightwaves = brighter image
2 out-of-phase lightwaves = arker image
2 bring both lightwaves together = phase-shift =increase contrast
2 ) t!pes3 phase-contrast 4 D5C
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Phase contrast microscope
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Phase contrast
2 provie greater resolution of internal structures
2 simplest phase microscope
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DIC microscope
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DIC
2 ifferential interference contrast
2 use prisms to split light beams into their
component wavelengths = increase
contrast
2 D
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lectron Microscopes
!" Types#
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lectron microscopes
2 use electrons instea of light
2 can resolve own to 1nm
2 T7 4 87
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lectron Microscopes
!" Types#
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TM
2 ) images w views of internal structures
2 works b! slicing specimen
2 stain w uranium salts to increase contrast
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SM
2 D images of e+ternal structures
2 paint surface of ob"ect w heav! metal
2 view the surface
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Staining
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Staining
2 increases contrast an resolution of a
specimen
2 !es stick through charge
chromophores, positivel!-charge !es
stick to the negativel! charge surfaces
most cells have
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$nionic and Cationic chromophores
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$nionic and Cationic
chromophores2 aciic !es contain anionic chromophores
2 less commonl! use
2 basic !es contain cationic chromophores
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Simple stains
!" types#
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Simple stains
2 irect an inirect
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Direct stains
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Direct stains
2 simple stains w basic !es 'such as C:
or meth!lene blue(
2 coating w positivel!-charge !e on
negativel!-charge surface of the cell
2 stains the organism
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Indirect stains
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Indirect stains
2 negative stains 'such as eosin(
2 slathering the slie w negativel!-charge
!e
2 stains aroun the organism
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Differential stains
!% types#
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Differential stains
2 ;ram
2 nospore
2
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&ram Stain
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&ram Stain
2 use to istinguish thickness of ; la!er2 ;ram positive = thick ; la!er 'purple(
;ram negative = thin ; la!er 'pink(
1$ smear 4 heat fi+
)$ primar! stain - basic !e 'C:(
$ morant - bins basic !e to cells '5(
>$ ecolori?er - thick ; la!er harer to remove'
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ndospore Stain
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ndospore stain
2 stains resting bo! of eh!rate DA< w protectivecoats
2 'enospores cannot be staine b! normal staining
proceures(
1$ smear an heat fi+
)$ primar! stain 'malachite green(
$ steam to push primar! stain pass the coat an into theenospore
>$ ecolori?e w water - enospore remains green
@$ counterstain 'safranin( - cell bo! now pink
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$cid-fast Stain
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$cid-fast stain
2 use on ;B cells with thick ; la!er an wa+! lipi cell wall
1$ smear an heatfi+
)$ primar! stain 'carbolfuschin($ heat 'steam( to push !e into wa+! cell wall 'carbolfuschin
issolves in wa+(
>$ ecolori?e w alcohol 4 /Cl 'if !e fastens to cells = aci-fast = pink
if !e washes off easil! = cell lacks wa+es = clear(@$ counterstain 'meth!lene blue( to stain bleache# non aci
fast cells
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'egati(e !Capsule# Stain
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Capsule layer
2 protein-sugar la!er outsie the cell wall
2 uncharge
2 use a mi+ture of a irect an an inirect stain
1$mi+ cells w congo re 'negative# aciic stain(
)$ let the smear air r!$ floo cells w basic fuschin !e 'sticks to ; la!er(
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@ main elementsthat make
up most of a cells mass Bfunctions
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P)'C*
2 C - backbone off organic molecules
2 / - organic molecules an water, /B release
b! acis2 - , /- release b! bases
2 A - amino acis# proteins# nucelic acis
2 - nucleic acis# atp# storage# transfer
membranes2 '8( - proteins
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5mportance of minor an
traceelements
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Minor + trace elements
elements that are re*uire in small 'trace, 1E(
amounts for proper growth
7inor elements3 8# F# 7g# Ca# Aa# Ge# Cl
Trace elements3 7n# Co# Hn# Cu# 7o
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Co(alent ,onds
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Co(alent ,onds
the sharing of a pair of electrons b! two
atoms
inclues / boning
olar covalent3 charge ue to une*ual sharing
of electronsAonpolar covalent3 atoms share electrons
e*uall! = uncharge
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*ydrogen ,onds
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* ,onds
2 / bone to a highl! A atom such as G# 5#
Ir# Cl# etc$2 e+plains wh! water is cohesive
2 structure2
weak, re*uires less energ! to break2 cells = J0E water = ver! important2 allows signaling to take place
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Ionic ,onds
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Ionic ,onds
2 two atoms e+hange electrons
2 usuall! between a metal an a nonmetal
2 charge
2 salts
2 issolvable b! water usuall!
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*ydrophylic (s *ydropho,ic
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*ydrophilic (s *ydropho,ic
2 /!rophilic3 water-loving, issolvable b!
water
2 /!rophobic3 water-fearing, ATissolvable b! water
2
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$cids (s Bases
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$cids (s Bases
2
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Cations (s $nions
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Cations (s $nions
2 caTions
2
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Cations (s $nions
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Cations (s $nions
2 caTions
2
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)rganic (s Inorganic
compounds
)rganic (s Inorganic
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)rganic (s Inorganic
compounds
2 5norganic3K compouns wo C e+cept C)K water# molecules# metal ions# aci
K compse )E of an organisms mass
2 rganic3K compouns w C mae b! cells e+cept C
an C)K compose appro+$ 9ME of cells
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*ydrocar,ons
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*ydrocar,ons
2 h!rophobic chains of C an /
2 issolve an react through the aition offunctional groups
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.unctional groups
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.unctional groups
2 allow certain molecules 'such ash!rocarbons( to react with each otherK N/
K NA/)K NC/K 8/K
N as polatir! ue to their electronegativit!
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Polymers (s Monomers
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Polymers (s Monomers
2 7onomers3 basic builing blocksK"oin together via eh!ration s!nthesis 'water is
release in the reaction# re*uires energ!(
2 ol!mers3 chains of monomersK break own via h!rol!sis 'water is a reactant#
releases energ! an monomers(
2 7ake up carboh!rates# proteins# nucleic acis#an lipis
7 7 h i
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7onomer3 7onosaccharie