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Microbiology > Day 4: Microscopy and Staining > Flashcards

Day 4: Microscopy and Staining Flashcards

1
Q

What are the 3 properties of light

A

1) wavelength
2) amplitude
3) frequency

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2
Q

Define wavelength

A

most important aspect

1 peak to the peak ( distance)

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3
Q

Define amplitude

A

height of peak

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4
Q

Define reflection

A

wave bounces off

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5
Q

Define absorbance

A

wave is captured

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6
Q

Define transmission/transmittsnce

A

wave travels through

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7
Q

Define resolution

A

ability of the lenses to distinguish fine detail and structure
( ability of the lenses to distinguish two parts that are a specified distance apart)

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8
Q

Define refraction

A

light wave changes direction as they enter a new medium

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9
Q

Define refractive index

A

a measure of the light bending ability of a medium

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10
Q

Define lenses

A

light passes through concex lens

refracted to a focal point

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11
Q

What did Antoine Van Leewenhoek contribute to microschopes

A

simple microscopre

animalcules

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12
Q

What did Robert Hooke contribute to early microscopes

A

compound microscope

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13
Q

Define light microscope

A

any kind of microscope that uses visible light to observe specimens

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14
Q

What are the different types of light microscopy

A 
Brightfield
Darkfield
phase-contrast
differential interference contrast ( DIC) 
fluorescence microscopy
confocal
two-photon
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15
Q

Describe the bright field microscopy

A

compound microscope
dark image on bright background
monocular or binocular

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16
Q

Define magnification

A

ability to enlarge images

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17
Q

Define total magnification

A

objective lens( 10x) x ocular lens( 4x, 10x,40x,100x)

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18
Q

Define resolution

A

ability to tell that 2 separate points or objects are separate

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19
Q

What are 2 factors affecting resolution

A

wavelength

numerical aperture

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20
Q

How do we prevent refraction in compound microscope

A

use of immersion oil ( light unrefracted)

w/o immersion oil ( most light is refracted and lost)

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21
Q

Define contrast

A

visible difference between the parts of a specimen

use different dyes and stains

22
Q

What are the 2 types of preparation

A

1) wet mount

2) fixed preparations

23
Q

Describe the wet mount technique

A

specimen is placed on the slide in a drop of liquid

24
Q

Define staining

A

coloring microorganisms with a dye to increase contrast

25
Define smear
thin film of a material containing microorganisms spread over a slide
26
Define fixed preparations
attaching cells to the slide | kills cells
27
What are the 2 types of fixed preparations
1) heat fixing | 2) chemical fixing
28
Define heat fixing
smear is prepared and briefly heated
29
Define chemical fixation | what chemicals? What is it used for? How does it work?
acetic acid, ethanol,methanol,formaldehyde mainly used for tissues denatures the proteins
30
What ions are consist in staining
stains are salts composed of positive and negative ions
31
What is chromophore
positive or negative ions that stain positively charged chromophore is a basic dye negatively charged chromophore acidic dye
32
What are examples of positive basic dyes(5) | What do they dye? cell or background? why?
cation in this basic dye is attracted to the negatively charged bacterial cell basic fuchsin, crystal violet, malachite green, methylene blue, and safranin
33
What is counterion
uncolored staining
34
What are examples of acidic dye ( 3) | What does the dye stain? Why does it stain this way?
negatively charged chromophore ex. acid fuchsin, eosin,rose bengal stain is repelled by cell wall so don't stain cell wall, instead dyes the background dye is negative and bacteria is negative so repel
35
What are the 2 types of stain
positive and negative
36
Define positive stain
staining the cells or organisms being observed | most commonly used
37
Define negative stain | What are we observing with this?
staining the background instead of the cell | valuable for observing overall cell shapes,size, and capsule
38
What are the 2 types of staining technique
simple staining | differential staining
39
What is simple staining | what are 4 examples
`primary purpose is to highlight the entire microorganism so that cellular shapes and basic structures are visible ex.methylene blue, carbolfuchsin, crystal violet,safranin
40
define differential staining
distinguishes organism based on their interactions with multiple stains most common stains caused for bacteria are gram stain and acid fast stain
41
What are the 3 differential staining techniques
gram staining acid-fast staining endospore staining
42
Describe gram staining
- classifies bacteria into gram-positive or gram-negative -based on the difference in cell wall composition ( gram positive: thick peptidoglycan cell wall) ( gram-negative: thin peptidoglycan cell wall)
43
Describe acid fast staining
differentiate gram positive bacteria with waxy lipid ( mycolic acid) bound yo peptidoglycan
44
What stain is used in acid fast staining
carbolfuchsin
45
What are the 2 techniques of acid fast staining
Ziehl-Neelsen and Kinyoun Technique
46
What bacteria can we use acid fast staining for
genus: bacteria mycobacterium genus: nocardia
47
Describe the endospore staining
primary stain: malachite green counterstain: safranin if stained with malachite green will have endospores
48
What is the method called in endospore staining
Schaeffer-Fulton method
49
What bacteria can we use endospore staining for
bacillus and Clostridium
50
Describe the capsule staining
capsule protective outer gelatinous covering negative staining ( dye as white outline of bacteria) stains used are India ink or nigrosin
51
Describe flagella staining
flagella-tail like structure used for locomotion | uses a mordant + pararosaniline or carbolfuchsin

Microbiology (16 decks)

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