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INTRO TO HISTO > ALL THE THINGS > Flashcards

ALL THE THINGS Flashcards

1
Q

What is another name for light microscopy?

A

bright field

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2
Q

Describe dark field microscopy?

A

uses scattered or refracted light from the specimen viewed on a dark background

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3
Q

What is phase or differential interference contrast?

A

Uses different refractive indices within a specimen for staining

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4
Q

What is another name for phase or differential interference contrast?

A

Nomarski

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5
Q

What is the Nomarski technique used to view commonly?

A

living cells

Example is cheek cells

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6
Q

What is fluorescence microscopy?

A

fluorescent molecules emit light at different wavelengths when exposed to UV light

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7
Q

What is confocal laser scanning microscopy?

A

uses a computer with a scanning laser beam to produce high res 3D images

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8
Q

What are the two types of electron microscopy?

A

transmission and scanning EM

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9
Q

Describe TEM

A

electrons pass through a section and are absorbed at different rates by variations in density in the tissue

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10
Q

Describe SEM

A

electrons reflected from the surface of a specimen

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11
Q

What are some downsides that keep us from using EM more often?

A

small sample size, harsh specimen prep, cumbersome and expensive, specimen has to be in a vacuum

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12
Q

What are the two lenses associated with light microscopy?

A

Objective and Ocular lens

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13
Q

What are the two stages of magnification?

A

first the objective lens then the ocular lens

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14
Q

What is the magnification range for the objective lens?

A

10X - 100X

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15
Q

What is the magnification range for the ocular lens?

A

8X - 10X

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16
Q

What is the diameter of a red blood cell?

A

approx 7.5 micrometers in diameter

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17
Q

How thick is the typical plasma membrane?

A

approx 10 nm thick

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18
Q

What is resolution?

A

the smallest distance between two items that are discernable as separate objects

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19
Q

Objects larger than __ can be distinguished as two separate objects

A

0.2 micrometers

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20
Q

What is the upper limit of magnification?

A

1000X

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21
Q

Increases in magnification are only valuable when accompanied by what?

A

adequate resolution

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22
Q

What are the 5 steps for preparing a tissue for microscopy?

A

fixation, embedding, sectioning, staining, mounting and coverslipping

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23
Q

What is the first thing you do when preparing an embedded tissue for microscopy?

A

dehydration of fixed tissue blocks in increasing concentrations of ethanol

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24
Q

What do you do after dehydrating your embedded tissue?

A

infiltrate it with organic solvent (Xylene)

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25
Q

After infiltrating with an organic solvent, what do you do?

A

embed the tissue in paraffin wax (or whatever you want)

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26
Q

After embedding in paraffin wax, what do you do?

A

section thinly in a microtome

27
Q

After sectioning in a microtome, what do you do with your sections?

A

mount on glass micriscope slides

28
Q

After mounting the sections on a slide, what do you do?

A

de-wax sections with xylene (organic solvent you initially used)

29
Q

After de-waxing, what do you do to your sample?

A

rehydrate with decreasing conentrations of alcohol

30
Q

After rehydrating your sample, what do you do?

A

stain in aqueous solutions of histological dyes

31
Q

After staining your sample, what do you do?

A

dehydrate again in alcohol series

32
Q

After dehydrating your now stained sample, what do you do?

A

further clear (de-wax) in xylene (or whatever miscible organic solvent you use)

33
Q

Finally, after clearing your sample, what do you do?

A

coverslipping with mounting medium

34
Q

Where are tissues prepped via cryo-sectioning commonly in order to obtain immediate biopsy results?

A

Hospitals

35
Q

Rapid freezing of tissues at very low temperatures prevents what?

A

destruction of proteins and enzymes

36
Q

What field of cytochemistry is cryo-sectioning commonly used?

A

immunocytochemistry

37
Q

What does freezing a tissue allow us to do?

A

hardens the tissue for thin sectioning

38
Q

In order to reduce artifacts in cryo-sectioned tissues, what are they stained with?

A

alcohol or other organic solvents

39
Q

List the purposes or benefits of fixing tissues for microscopy

A

quickly terminate metabolic processes, prevents autolysis of cells and tissues by enzymes; kill any pathogenic organisms

40
Q

What are the two common chemical fixtatives used for fixation?

A

formalin and alcohol

41
Q

What is formalin? (Chemically)

A

37% aqueous solution of formaldehyde

42
Q

What does formaldehyde do when we use it to fix tissues?

A

forms cross linkages (covalent bonds) between adjacent amino acid groups of proteins

43
Q

Which amino acid group is particularly affected by formaldehyde?

A

lysine groups

44
Q

Why do we add alcohol after treating with formalin?

A

alcohol denatures proteins via dehydration and precipiration

45
Q

What is another common duo of fixatives used for fixation?

A

glutaraldehyde and picric acid

46
Q

When embedding a tissue, why should your organic solvent be miscible with paraffin wax?

A

we don’t want something that will react and start forming unnecessary side products with the wax

47
Q

What kinds of knives are commonly used to section embedded tissues?

A

steel, glass or diamond

48
Q

What do we use to section embedded tissues? (What is the machine called)

A

Microtome

49
Q

How thick are embedded tissues generally cut?

A

5-20 micrometres

50
Q

After sectioning, what do we use to stick the pieces on our slides?

A

egg albumin or some other sticky media

51
Q

What is the most common staining technique used?

A

Haematoxylin and Eosin staining

52
Q

What kind of cellular components does haematoxylin bind and what color does it stain?

A

binds acidic compounds (nucleic acids). stains blue

53
Q

What stain is used for basic compounds like proteins?

A

Eosin and it stains red

54
Q

Can mucus be stained? If not how will it appear under the microscope?

A

No because it is relatively neutral and will appear colourless

55
Q

H&E stains together will stain what color?

A

Purple. Used for the nucleus and the cytoplasm too.

56
Q

What is a metachromatic stain?

A

the color of the stain changes once it binds certain tissues

57
Q

What kind of stain would you use for carbohydrate rich compounds like glycogen?

A

Periodic Acid-Schiff stain (Pinkish-Purple)

58
Q

What kind of stain is used for elastic tissues?

A

Weigert’s elastic tissue stain (Purple)

59
Q

What two stains are used for neural tissue stain?

A

Nissl and myelin stains

60
Q

What are commonly used Nissl stains?

A

Cresyl Fast Violet

61
Q

What are commonly used myelin stains?

A

Luxol Fast Blue

62
Q

What is commonly stained by Nissl stains?

A

ER and other things made up of lots of RNA

63
Q

Myelin stains are used to visualize what?

A

fiber bundles (myelinated nerve fibers)

64
Q

What are some common artifacts of tissue preparation?

A

shrinkage due to alcohol fixation, artificial spaces created by shrinkage, loss of substances due to fixation, ice crystal formation

INTRO TO HISTO (1 decks)

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