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Histology > Ch 1 > Flashcards

Ch 1 Flashcards

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1
Q

Histology

A

the study of the microscopic structures of tissues and organs

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2
Q

What is the goal of histology?

A

to correlate structure with function

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3
Q

What diverse methods does histology utilize?

A

light, virtual, and electron microscopy

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4
Q

What is the first step to tissue preparation?

A

Fixation

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5
Q

What is fixation?

A

the first step in tissue preparation to preserve structure and prevent degradation

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6
Q

What is used in fixation?

A

formalin

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7
Q

what is formalin

A

a common fixative

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8
Q

What is the second step in tissue preparation

A

embed the specimen in paraffin

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9
Q

what does embedding the specimen in paraffin require?

A

dehydration and clearing, to permit sectioning

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10
Q

What is the third step in tissue preparation?

A

sectioning

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11
Q

what is sectioning?

A

slicing the embedded tissue using a microtome

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12
Q

what is the fourth step in tissue preparation

A

staining

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13
Q

Why is staining necessary

A

because paraffin sections are colorless

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14
Q

What are two most common staining methods?

A

Hematoxylin and eosin (H&E)

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15
Q

What does hematoxylin do?

A

stains nuclei blue

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16
Q

What does eosin do?

A

stains cytoplasm pink

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17
Q

What is the fifth step in tissue preparation?

A

mounting the stained section on a glass slide with mounting medium

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18
Q

What are the steps of tissue preparation?

A
  1. Fixation
  2. Embedding in parafin
  3. Sectioning
  4. Staining
  5. mounting
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19
Q

Why are other fixatives sometimes used?

A

to retain component not preserved by formalin

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20
Q

What are the two the fixatives mentioned?

A
  1. heavy metals like osmium tetroxide
  2. freezing
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21
Q

What are heavy metals such as osmium tetroxide are used for?

A

to perserve membrane structures

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22
Q

What is freezing sections used for?

A

to retain neutral lipids

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23
Q

What would other staining procedures be used?

A

to display specific structural components

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24
Q

What are the two other staining techniques mentioned?

A
  1. orcein and resorcin-fuchsin
  2. Silver impregnation
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25
What is orcein and resorcin-fuchsin are used for?
staining procedure for elastic material
26
What is silver impregnation is used for? | 2 things
Is is a staining method for; - reticular fibers - basement membranes
27
what do all the procedures provide information about?
cell function and extracellular components
28
What might procedures be based on? | 3 things
1. specific dying binding 2. fluorescent dye labeled antibodies 3. inherent enzymatic activity
29
What is autoradiography?
uses radioactively tagged precursors to localize molecules
30
What is the chemical basis of staining?
acidic and basic dyes are commonly used
31
What is an example of an acidic dye?
eosin
32
What does an acidic dye like eosin do?
carries a negative charge and reacts with cationic components
33
What is an example of a basic dye?
hematoxylin
34
What does a basic dye like hematoxylin do?
carries a positive charge and reacts with anionic components
35
What are other examples of basic dyes?
1. methyl green - green 2. methylene blue - blue 3. pyronin G - red 4. toluidine blue - blue
36
What are other examples of acidic dyes?
1. acid fuchsin - red 2. aniline blue - blue 3. eosin - red 4. orange G - orange
37
what is metachromasia?
a color shift seen with certain basic dyes
38
what does the periodic acid-Schiff (PAS) reaction do?
stains carbohydrates and carbohydrate-rich molecules
39
What does feulgen stain do?
sues the Schiff reagent to stain DNA
40
What can be used to confirm the identity of stained material?
enzyme digestion
41
what does enzyme histochemistry do?
localizes enzymes in tissue sections
42
what does immunocytochemistry do?
uses the specificity of antigen-antibody reactions to locate molecules
43
what techniques does immunocytochemistry use?
direct and indirect
44
What happens in direct immunofluorescence?
1. a fluorochrome-labeled primary antibody reacts with a specific antigen within the tissue sample 2. labeled structures are then observed in the fluorescence microscope in which an excitation wavelength (usually UV light) triggers the emission of another wavelength 3. the length of the wavelength depends on the nature of the fluorochrome used for antibody labeling
45
What happens in indirect immunofluorescence?
1. specific primary antibodies react with the antigen of interest 2. the secondary antibodies, which are fluorochrome labeled, react with the primary antibodies 3. the visualization of labeled structures within the tissues is the sam in both methods and requires the fluorescence microscope
46
What is hybridization used for?
to legalize mRNA or DNA using a complementary nucleotide probe
47
what is a type of hybridization?
fluorescence in situ hybridization (FISH)
48
What does FISH do?
uses fluorescent dies with probes
49
What does autoradiography use?
a photographic emulsion to localize radioactive material
50
What does expansion microscopy do?
ExM physically expands specimens to improve resolution - specimens are infiltrated with swellable polymers
51
What are the steps in preparation of a specimen for expansion microscopy?
1. Fixation - cell in formaldehyde 2. Anchoring - add anchoring reagents (acryloyl groups) that bind to proteins or other molecules of interest and molecular probes conjugated with fluorescent labels, infuse with superabsorbent (sodium acrylate) 3. Genlation - sodium acrylate - triggers development of a sense three-dimensional hydrogel polymer matrix 4. mechanical homogenization - allows the cells to break open using ultrasonic homogenizer (creating proteinases), 5. Expansion - the specimen is now embedded in a hydrogel matrix, is ready for physical expansion by the addition of water. add water to expand (proportionally in all directions making cell bigger) how does expansion work - th proteins of interest remain connected to the expanded polymer network which pulls them apart (aprox. 4.5 times bigger) 6. examination - fluorescence microscope
52
What are the types of microscopy?
1. light microscopy 2. electron microscopy 3. atomic force microscopy (AFM) 4. virtual microscopy
53
What is light microscopy?
uses a light source to magnify images
54
What is resolving power?
the ability to produce separate images of closely positioned objects
55
that is the most common type of light microscope?
the bright field microscope
56
What are the other types of optical systems?
1. phase contrast 2. dark field 3. fluorescence 4. confocal 5. ultraviolet microscopy
57
What is electron microscopy?
uses electrons to produce high resolution images
58
What are the types of electron microscopes?
transmission electron microscopy (TEM) and scanning electron microscope (SEM) - in both of these specimens need to be inserted into a high-vacuum environment
59
what is atomic force microscopy (AFM)?
uses a probe to scan the specimen's surface at high resolution - a tip moves up and down with the surface to produce an image
60
What does virtual microscopy do?
provides a digital alternative to examining glass slides using a light microscope

Histology (5 decks)

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