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Microbiology Chapter 2
Microscopy and Cell Components
263
Microbiology
12th Grade
01/31/2011

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Term
Micrometer (µm)
Definition
10^-6 m
Term
nanometer (nm)
Definition
10^-9 m
Term
How many zero's will there be after the decimal when converting 21.5 nm to m?
Definition
7: 0.0000000215
Term
Total magnification
Definition
objective * Ocular
Term
Resolution
Definition
Distance apart needed to see 2 points as separate (ability to see fine detail)
Term
With respect to resolution in microscopes, which is better, a small number or large one?
Definition
small
Term
With respect to resolution in cameras, which is better, a small number or large one?
Definition
large
Term
Which is better for seeing small organisms, a light or election microscope?
Definition
electron microscope
Term
How do you calculate the resolving power?
Definition
Wavelength divided by (2 * NA)
Term
NA
Definition
Numerical Aperature
Term
What does numerical aperature (NA) depend on? (2)
Definition
-Refractive index of material between lense and slide
-The angle of most divergent light ray that can enter that objective
Term
What type of material will we use for our refractive index in class? (2)
Definition
oil or air
Term
What are the two (three) ways of improving resolution?
Definition
-Decrease wavelength
-Use oil with 100x objective
(-increase N.A.)
Term
How would one decrease wavelength when they are trying to improve resolution?
Definition
electron vs. visual light
Term
How does one improve the conditions but NOT resolution? (2)
Definition
-contrast by staining
-light adjustment
Term
If you use a low objective, should you use a high or low light?
Definition
low light
Term
If you use a high objective, should you use a high or low light?
Definition
high light
Term
What does a compound microscope mean?
Definition
two lenses
Term
[image]
Definition
adjustment knobs
Term
[image]
Definition
Arm
Term
[image]
Definition
Base
Term
[image]
Definition
Body Tube
Term
[image]
Definition
Condenser
Term
[image]
Definition
Iris Diaphragm
Term
[image]
Definition
Light
Term
[image]
Definition
Mechanical stage knobs
Term
[image]
Definition
Occular Eyepiece
Term
[image]
Definition
Objective Lenses
Term
[image]
Definition
Stage
Term
Occular (eyepiece)
Definition
A magnifying lense (usually 10x)
Term
Arm
Definition
carrying/support
Term
Stage
Definition
Platform that holds the microscope slide in position
Term
Course/fine adjustment knobs
Definition
Used for focusing the specimen;turning he knobs changes the distance between the objective lens and the specimen
Term
base
Definition
carrying/support
Term
body tube
Definition
contains mirrors and prisms that transmit the image to the ocular lens
Term
objective lenses
Definition
primary lenses that magnify the specimen
Term
Condenser
Definition
A lens system that condenses light before it passes through the specimen
Term
Iris diaphragm
Definition
controls the amount of light entering the condenser
Term
mechanical stage knobs
Definition
used to move the slide
Term
light
Definition
source of illumination
Term
Light Brightfield microscope:
1. Background
2. Resolution
3. Magnitude
Definition
1. Bright
2. 0.2µm
3. 2000x
Term
light Darkfield:
1. Background
2. Resolution
3. Magnitude
4. Microbes
Definition
1. dark
2. 0.2µm
3. 2000x
4. clear
Term
light, fluorescent
1. Background
2. Resolution
3. Magnitude
4. Microbes
Definition
1. dark
2. 0.2µm
3. 2000x
4. bright and fluorescing
Term
What is the light, brightfield microscope enhanced by? (3)
Definition
-stains
-oil with 100x lens
-Diaphragm and light
Term
What is the light, darkfield microscope enhanced by? (3)
Definition
NONE
Term
What is the light, fluorescent microscope enhanced by? (2)
Definition
-Fluorescent antibody dyes
-Fluorescent dye on antibody (specific) to microbe antigen, microbe fluoresces.
Term
What are the uses of the light, brightfield microscope? (3)
Definition
-Live specimens (unstained)
-stained (dead) specimens
-bacteria, protozoa, fungus, multicellular animal parasites eggs
Term
What are the uses of the light, darkfield microscope? (1)
Definition
live microbes:syphillis
-we can't stain, too thin
Term
What are the uses of the light, fluorescent microscope? (2)
Definition
-when immediate diagnosis needed
-When cultures aren't available, or take too long.
Term
What is the light, fluorescent microscope used to test
Definition
TB, rabies, syphillis, anthrax
Term
Electron,scanning
1. Resolution
2. Magnification
Definition
1. 20Nm
2. 10,000x
Term
Electron,transmission
1. Resolution
2. Magnification
Definition
1. 2.5 Nm
2. 100000x
Term
What is the electron scanning microscope enhanced by?
Definition
Nothing
Term
What is the electron transmission microscope enhanced by?
Definition
Stain w/ + salt of heavy metal (barium)
Term
What are the advantages (1) of the electron scanning microscope?
Definition
3-d
Term
What are the advantages (1) of the electron transmission microscope?
Definition
high resolution and magnitude
Term
What is the disadvantage to all e- scopes
Definition
artifacts due to killing, and fixing under vacuum.
Term
artifacts
Definition
problem with all scopes, distorations to microbes.
Term
What is the disadvantage to the electron transmission microscope?
Definition
need THIN slice as e- can't penetrate
Term
What are the uses of an electron scanning microscope? (1)
Definition
surface structures- eukaryote to virus
Term
What are the uses of an electron transmission microscope? (4)
Definition
-Virus particles
-bacterial flagella
-internal cell structures
-protein molecules
Term
What are the steps to slide prep for basic stains? (4)
Definition
1. Smear
2. Air dry
3. Fix
4.Stain
Term
Fix (a slide)
Definition
attach to a slide (won't come off)
Term
What is used to fix a slide?
Definition
alcohol or heat (hopefully preserves with little distortion)
Term
Does fixing kill the microbe?
Definition
yes
Term
Basic dye: positive or negative stain?
Definition
positive
Term
How do poitive stains/basic dyes work?
Definition
The postive dye is attracted to the negative cell wall., therefore stains the cell.
Term
What are examples of basic dyes?
Definition
Crystal violet, methylene blue, safranin (red) malachite green
Term
Are acidic dyes positive or negative stains?
Definition
negative
Term
What type of microscope is a negative stain similar to?
Definition
light, darkfield scope.
Term
How does an acidic dye work?
Definition
Negative dye repels negative cell wall. Stains backround instead of microbe.
Term
Why is a negative dye used?
Definition
for cell shape and size, to detect capsules
Term
What are the advantages to using an acidic dye?
Definition
less distotion (no heat fixing, stain doesn't enter)
Term
What are the examples of acidic dyes?
Definition
Acid Fuchsin, nigrosin
Term
capsule
Definition
gelatinous covering on outside, makes microbe more deadly
Term
mordant
Definition
substance used to cause more intense staining
Term
Is a mordant a stain?
Definition
no
Term
What is a mordent used for?
Definition
to enhance a stain
Term
Simple stain
Definition
use of simple basic dye
Term
What color are microbes in a simple stain?
Definition
all one color
Term
What is a simple stain used for?
Definition
morphology (shape, arrangement, size)
Term
Differemtial stain
Definition
use of greater than two stains to distinguish groups of bacteria
Term
What are examples of a differential stain?
Definition
gram stain, acid fast stain
Term
gram stain
Definition
differential due to cell wall differences
Term
G.P.
1. Postive or negative?
2. Color?
3. Which stain?
4. Penicilin?
Definition
1. Positive
2. Purple
3. Retains primary stain
4. susceptible to penicillin
Term
G.N.
1. Postive or negative?
2. Color?
3. Which stain?
4. Penicilin?
Definition
1. negative
2. red
3. accepts counterstain
4. Resistant to penicilin
Term
What are staining problems for gram stains? (5)
Definition
-need young cultures
-some bacteria stain poorly
-Decolorization timing is critical
-Potential artifacts
Term
what are potential problems with all stains? (2)
Definition
-decolorization timing is critical
-Potential artifacts
Term
Why do you need young cultures?
Definition
holes in cell wall (old skin)could let purple out
Term
Any time you add dye with heat, what are you staining?
Definition
an endospore
Term
Does a darkfield microscope use light?
Definition
yes
Term
Does an electron scanning microscope use light?
Definition
no
Term
Does an electron transmission microscope use light?
Definition
no
Term
What is the order of chemicals in the gram staining process? (4)
Definition
1. Crystal Violet
2. Iodine
3. Alochol
4. Saphranin
Term
What color is the GN and GP after the crystal violet?(Gram Stain)
Definition
GN:Purple
GP:Purple
Term
What color is the GN and GP after the iodine? (Gram Stain)
Definition
GN: Purple
GP:Purple
Term
What color is the GN and GP after the alocohol? (Gram Stain)
Definition
GN:Clear
GP: Purple
Term
What color is the GN and GP after the saphranin? (gram Stain)
Definition
GN: Red
GP: Purple
Term
What is the point of crystal violet in a gram stain?
Definition
To stain all of the cells one color
Term
What is the point of iodine in a gram stain?
Definition
Mordant
Term
Is iodine a stain?
Definition
No its a mordant
Term
What is the point of alochol in a gram stain?
Definition
dehydrates cell wall
Term
What is the point of safranin in a gram stain?
Definition
to give contrast
Term
In an acid fast stain, the primary stain is?
Definition
red
Term
In an acid fast stain, the primary stain color is...?
Definition
red
Term
In an acid fast stain, the scondary stain color is?
Definition
Blue
Term
In an acid fast stain, is a positive result the color of the primary stain or the scondary stain?
Definition
red, the primary stain
Term
In an acid fast stain, is a negative result the color of the primary stain or the secondary stain?
Definition
Secondary stain, blue
Term
In an acid fast stain, why does the poitive stain retain its color?
Definition
the wax in the cell wall keeps the primary stain
Term
What is the acid fast stain used for?
Definition
To ID mycobacterium species, TB (tuburculosis, leporasy, avium)
Term
Capsule
Definition
a gelatinous covering on outside of bacteria to help it evade immune system. The bacteria secretes like slime.
Term
How does one perform a capsule stain? (3steps)
Definition
1. Use positive stain to stain bacteria
2. Negative stain to stain background
3. clear halo of capsule is left between the stains
Term
What can be a problem with a capsule stain?
Definition
the capsule can wash off as it is soluble in water.
Term
Endospore
Definition
resistant, dormant resting structure to protect microbe from adverse conditions.
Term
Is an endospore used for reproduction?
Definition
no
Term
What is used to determine endospore species?
Definition
its position: terminal, subterminal, and central
Term
How do you stain an endospore?
Definition
use heat to force dye into stain resistant endospore.
Term
What are the three positions an endospore can be in?
Definition
terminal, subterminal, central
Term
[image]
Definition
Capsule
Term
[image]
Definition

1. central

2. terminal

3. terminal

4.central

5. terminal

 

Term
Flagella
Definition
whip-like structure for mobility
Term
What, in flagella, is used to id bacteria?
Definition
-number and arrangement
-different proteins on flagella
Term
can coccus bacteria have flagella?
Definition
no
Term
Can bacillus have flagella?
Definition
yes
Term
can spirals have flagella?
Definition
yes
Term
Do prokaryotes have a nucleus?
Definition
no, they have a nucleiod region but no membrane.
Term
Do prokaryotes have any membranous organelles?
Definition
no, but they do have ribosomes
Term
Do prokaryotes have DNA?
Definition
yes, 1 circular chromosome.
Term
Do prokaryotes have the same protein associated with the their DNA as eukaryotes?
Definition
no, eukaryotes have histone associated with their DNA. There is a different protein associated with the DNA of prokaryotes
Term
How do bacteria reproduce?
Definition
binary fision
Term
What is a bacteria's cell wall is made of?
Definition
peptidoglycan, murein
Term
What is an archea's cell wall made of?
Definition
they don't have any. If they do its pseudomurien.
Term
What is the purpose of a bacteria's cell wall? (3)
Definition
-maintains shape
-anchors flagella
-prevents rupturing due to osmotic pressure
Term
What is the clinical importance of a bacteria's cell wall? (2)
Definition
-Site of antibiotic activities
-Determines GN or GP
Term
What is a bacteria's cell wall made of?
Definition
peptidoglycan
Term
How does penicillin affect a gram positive cell wall?
Definition
Penicillin interferes with peptide links in GP=> weak cell wall=>water intake &lysis (increase in osmotic pressure)=>dies
Term
How many layers of cell membrane are in the GP?
Definition
1
Term
How many layers of cell membrane are in a GN?
Definition
2
Term
Does a GP cell wall have telchoic acid?
Definition
yes
Term
Does a GN cell wall have telchoic acid?
Definition
no
Term
Does a GP cell wall have two layers?
Definition
no
Term
Does a GP cell wall have two layers?
Definition
no
Term
Does a GN cell wall have two layers?
Definition
yes
Term
Which cell wall has a thick peptidoglycan layer, GN or GP?
Definition
GP
Term
Which cell wall has a thin peptidoglycan layer, GN or GP?
Definition
GN
Term
Which cell wall has lyposcpolysaccharides(LPS), GN or GP?
Definition
GN
Term
Does a GP cell have a thick peptidoglycan layer in their cell wall?
Definition
yes
Term
Does a GP have telchoic acid in their cell wall?
Definition
yes
Term
Which cell wall (GN or GP) secretes primarily extotoxins?
Definition
GP
Term
Do Gram Positive cells have a space between the layers of its cell wall?
Definition
No, there is only one cell wall...no space...
Term
Do GN cell walls have thick or thin layer of peptidoglycan?
Definition
thin
Term
Do GN cell walls have trichoic acids?
Definition
no
Term
Do GN cell walls have an outer wall embrane?
Definition
yes
Term
What is a GN's outer wall membrane function?
Definition
evades phagocytosis, it is a barrier to penicillan and enzymes
Term
porins
Definition
allows certain things to get through, opening
Term
How do things get through a GN's cell wall?
Definition
porins
Term
What do the lypopolysacharides in a GN cell wall do?
Definition
secrete endotoxin which can lead to a fever and shock.
Term
Does a GN cell wall have a space between its two layers?
Definition
yes, periplasm
Term
Where is the peptidoglycan found in a GN cell wall?
Definition
periplasm
Term
Periplasm
Definition
space between outerwall and plasma membrane, fluid, where peptidoglycan is
Term
What does the periplasm contain?
Definition
degradative enzymes
Term
What can make a GN appear GP?
Definition
don't decolorize enough
Term
What affect does alcohol have on GP in a gram stain?
Definition
dehydrates, increases density of thick wall, crystals can't leave
Term
What affect does alcohol have on GP in a gram stain?
Definition
disolves outer membrane and dehydration leaves a hole in thin peptidoglycan, crystals exit and sell colorless
Term
How can a GP falsely stain GN? (3)
Definition
Cell wall damaged due to:
-age
-heat damage during fixing
-overdecolorization
Term
What are the only type of bacteria able to make endospores?
Definition
GP rod/bacillus
Term
What are the only bacteria able to test positive in an acid fast stain?
Definition
GP rod/bacillus
Term
GP rod/bacillus only test poitive for (2)
Definition
-endospore stain: Bacillus & clostridium
-Acid Fast STain:Mycobacterium (TB)
Term
Lysozyme
Definition
digeestive enzyme in saliva, tears and mucus
Term
What type of bacteria is a lysozyme most effective against? (GN/GP)
Definition
GP:it hydrolyzed peptide bonds
Term
What type of bacteria is penicillin most affective against? (GN/GP)
Definition
GP: breaks down peptide bonds
Term
mycoplasma species
Definition
NO CELL WALL
Term
Name three species with an atypical cell wall/
Definition
Mycoplasma
-mycobacteria
-archea
Term
What in its plasma membrane prevents a mycoplasma from lysis (bursting)?
Definition
the high amount of sterolipid
Term
Mycobacteria
Definition
high wax in wall
Term
What will a mycobacteria stain if gram stain?
Definition
MUST ANSWER GP!!!!
-hard to stain, wax and water don't mix well)
Term
In an acid fast stain, what will mycobacteria be?
Definition
positive:TB
Term
In an acid fast stain, what will mycoplasma be?
Definition
negative...
Term
ARchea cell wall?
Definition
No cell wall or pseudomurein
Term
What is another term for a capsule?
Definition
Glycocalyx
Term
What is another term for glycocalyx?
Definition
capsule
Term
What is a capsule/glycocalyx made of? (2)
Definition
(sugar coat)
-EPS (Extra cellular polysaccharide) and polypeptide polymer
Term
What is the purpose of a glycocalyx/capsule?
Definition
-increased virulence, evade phagocytosis, adherence, dehydration protetion
Term
How would one stan a capsule/glycocalyx? Include stains used.
Definition
Negative stain with two dyes: Basic (poitive)stains organism, acidic(negative) stains background. Halo between is the capsule
Term
What is the purpose/function of flagella?
Definition
movement
Term
What is the purpose/function of axial filaments?
Definition
movement (corkscrew)
Term
What is the purpose/function of fimbrae?
Definition
adherence (to stick)
Term
What is the purpose/function of pilli?
Definition
transfer DNA (plasmids only) "sex pili"
Term
In what group of bacteria would we find flagella?
Definition
-bacillus
-spiral
Term
In what group of bacteria would we find axial filaments?
Definition
spirochetes
Term
In what group of bacteria would we find fimbrae?
Definition
Gram Negative
Term
In what group of bacteria would we find pili?
Definition
GN
Term
How do GP bacteria transfer plasmids?
Definition
NOT PILI
-sticky substance connecting cells long enough to transfer plasmid
Term
How do GN transfer plasmids?
Definition
sex pili
Term
monotrichous
Definition
1 at 1 pole
Term
amphitrichous
Definition
tuff (1 or more) at each pole
Term
lophotrichous
Definition
more than 1 at 1 pole
Term
peritrichous
Definition
all around
Term
location of Axial filaments on a cell
Definition
spiralled around cell within outer sheath (endoflagella)
Term
location of fimbrae on a cell
Definition
few to hundreds all around cell
Term
amount of pili on a cell
Definition
0-1 per cell
Term
External filamentous structures
Definition
-thin, extend from cell wall
-only in certain bacteria, not all
Term
do bacteria reproduce through pili?
Definition
no only binary fusion
Term
Endospore
Definition
protection from extreme conditions. Not reproduction.
Term
F- cell
Definition
doesn't have plasmid or sex pillus
Term
F+ cell
Definition
has plasmid and sex pilus
Term
Are there cilia on bacteria?
Definition
no
Term
taxis
Definition
movement in response to stimulous
Term
Chemotaxis
Definition
attracted to or try to get away from chemical
Term
phototaxis
Definition
attracted to or try to get away from light
Term
Endospore
Definition
resistant "resting" structures to survive adverse conditions
Term
What can make endospores?
Definition
GP RODS
-bacillus and clostridium
Term
Sporultion/sporogenisis
Definition
-makes spores
-goes from active bacteria to dormant endospore
Term
Germanation
Definition
-goes from dormant endospore to vegatative (active) bacteria
Term
What are locations of endospores (3)
Definition
terminal,subterminal, central
Term
What can an endospore survive?
Definition
dehydration, heat, chemicals, improper canning(toxins=>food poisoning), etc.
Term
How does an endospore appear in a gram stain?
Definition
clear
Term
How is an endospore stain made?
Definition
-primary: basic stain with heat forces stain inro endospore
-rinse: removes stain from rest of slide/organism
-Counterstian: basic stain colors rest of organism
Term
Plasma membrane
Definition
-bilayer of phospholipids with protein
-selectively (semi) permeable
Term
What is an organism with a special plasma membrane?
Definition
mycoplasmas:contain cterol
Term
What can be effective in dimishing the plasma membrane?
Definition
alcohols, disinfectants, some antibiotics
Term
Diffusion
Definition
passive transport high to low concentration
Term
simple diffusion
Definition
goes right through the membrane
Term
fascilitated diffusion
Definition
involves a protein, does not cause energy usage
Term
osmosis
Definition
diffusion of water
Term
active transport
Definition
from low to high concentration. Requires ATP (energy) and protein
Term
What are the three osmotic enviroments?
Definition
isotonic, hypotonic, hypertonic
Term
Isotonic/isoosmotic solution
Definition
equal % total solute on both sides of membrane
-water moves in both directions
-Dynamic equilibrium (cell size constant)
Term
Hypotonic solution
Definition
-low % solute
-net water moves into the cell
-if no cell wall, or weakened, lysis occurs
Term
How does the cell wall help a hypotonic solution?
Definition
keeps cell from bursting
Term
How does the cell wall help a hypertonic enviroment?
Definition
it doesn't. Does not prevent water from leaving (can shrivel and die)
Term
Which is more detrimental for a cell? a hypotonic solution or hypertonic enviroment?
Definition
hypertonic enviroment
Term
Hypertonic Solution
Definition
-high % colute
-Net water movementout of cell
-plasmolysis can occur
Term
plasmolysis
Definition
cell shrivels and dies from lack of water.
Term
Chromatophores/thylakoids
Definition
photosynthetic structures
Term
Bacteria nucleoid/nuclear area
Definition
-no nuclear membrane
-1 circular chromosome w/out histones
Term
Plasmids
Definition
extrachromocomal DNA
Term
What is the benfitst of plasmids to the organisms?
Definition
helps survive
Term
What is the benefit of plasmids to us?
Definition
genetic engineering
Term
How many genes are in a plasmid? can it transfer between species?
Definition
4-5, yes
Term
How are plasmids transferred?
Definition
-Conjunction (transfer through direct contact
-GN sex pili
-GP sticky substance
Term
Are plasmids crucial for survival?
Definition
no
Term
What do people mean when they say plasmids are biotech?
Definition
used for genetic manipulation
Term
Can an f- change to an f+?
Definition
yes
Term
Can an F+ change to an F-?
Definition
no
Term
S
Definition
Sedimentaion (how fast they go to bottom of tube (settle out))
Term
Ribosomes
Definition
protein synthesis
Term
What is a ribosome made of?
Definition
2 subunits protein and RNA
Term
Prokaryotic ribosome size?
Definition
70s (30s&50s subunits)
Term
Eukaryotic ribosome size?
Definition
80s (60s & 40s subunits)
Term
How do antibiotics attack prokaryotic ribosomes and not eukaryotic ribosomes?
Definition
Antibiotics attach to subunit of prokaryotic size
Term
Inclusions
Definition
reserve deposits
Term
How do inclusions help the cell
Definition
prevent increase in osmotic pressure
-help with water pull, decrease osmotic pull
Term
Why is an endospore stain needed?
Definition
Endospore and not inclusion both look clear, endospore stain, inclusion does not
Term
what are some inclusions? (3)
Definition
-sulfar, sucrose, air (buoyency)
Term
What are some prokaryotic structures?
Definition
flagella, fimbrae, plasmid, chromosome, inclusion, ribosome, plasma membrane, capsule
Term
What looks different in an eukaryote than a prokaryote?
Definition
-larger
-Nucleus (membrane bound organelles)
-If cell wall: algae:cellulose, Fungi:Chitin
Term
What groups have fimbrae?
Definition
GN
Term
Waht groups have flagella?
Definition
bacillus spirilum
Term
special GP rods?
Definition
-endospore stain: has endospore
-acid fast:test for wax
Term
Endosymbiotic theory
Definition
Eukaryotes evolved from sympbiotic prokaryotes liviing inside other prokaryotes
Term
What are 3 things that serve as evidence for the endosymbiotic theory?
Definition
-Organelle membranes are similar(phospholipid)like bacterial plasma membranes
-Choloplasts similar to photosynthetic prokaryotes
-Mitochondria and Chloroplasts
Term
What is so significant about mitochondria and cholorplast when dealing with the endosymbiotic theory?
Definition
-reproduce by binary fision
-Contains DNA (circular without histones) and 70s ribsomes
-Double membrane like prokaryotic plasma membrane
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