Microbiology Review 1

7/25/2019 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



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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

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Microbiology Review 1