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Fundamentals Of Medicine: Exam 1 > Microscopy And Histology > Flashcards

Microscopy And Histology Flashcards

1
Q

What is cytology

A

Study of cells

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2
Q

Define histology

A

Study of tissues

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3
Q

What are the two major types of microscopes?

A
  1. Light microscopes
  2. Electron microscopes
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4
Q

The ability to make small objects seem larger

A

Magnification

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5
Q

Discrimination between two objects that are close together

A

Resolution

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6
Q

The ability to perceive the presence of an object

A

Detection

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7
Q

Magnification without revealing more detail (every optical system has a finite limit of resolution/resolving power)

A

Empty magnification

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8
Q

The type of tissue preparation that takes several days but has the highest resolution

A

Paraffin-Embedded Sections

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9
Q

In Paraffin Embedded tissue preparation, what is the water in tissue replaced with?

A

Paraffin wax

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10
Q

What are the 6 steps of Paraffin Embedded tissue preparation

A
  1. Fixation: using formaldehyde
  2. Dehydration: using alcohols
  3. Clearing: using xylene
  4. Infiltration and embedding: using paraffin wax
  5. Sectioning: use microtome
  6. Staining
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11
Q

What type of tissue preparation is often used during surgery?

A

Frozen sections

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12
Q

What type of tissue preparation only takes a few minutes but has lower resolution

A

Frozen sections

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13
Q

What are the 4 steps of frozen section tissue preparation?

A
  1. Fixation: using acetone
  2. Embedding and freezing: using glycols and resin (removes water)
  3. Sectioning: using cryostat
  4. Staining
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14
Q

If tissues dye and are not fixed fast enough, what artifact can occur?

A

Post mortem degeneration, where cells are pulled away from connective tissue leaving open spaces

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15
Q

When does shrinkage occur?

A

When a tissue sample is not fixed fast enough

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16
Q

Which tissue preparation uses formalin for fixation?

A

Paraffin-embedded Sections

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17
Q

What is required to visualize colorless tissue and cells under a microscope?

A

Staining

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18
Q

What is the most common type of tissue staining?

A

Hematoxylin and Eosin (H&E)

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19
Q

What charge and color is Hematoxylin?
What does it bind to?

A

A blue dye with a positive charge. Binds to RNA and DNA (negative charged phosphate group). Binds to sulfate groups

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20
Q

Define basophilic

A

Tissue components that stains with basic dyes. Example: DNA and RNA

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21
Q

What color will the Nucleus stain using H&E

A

Blue

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22
Q

What components in tissue are basophilic?

A

Nucleus (DNA) and basal ER (RNA)

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23
Q

What color and charge is Eosin dye?

A

Eosin is a red, negatively charged dye

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24
Q

Is Eosin basic or acidic?

A

Acidic

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25
Q

Define acidophilic

A

Tissue components that stain with acidic dyes

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26
Q

What color would tissue components containing a NH3+ stain when using H&E

A

Red

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27
Q

What color would proteins stain when using H&E

A

Red (NH3+ group)

28
Q

What other components stain red in tissue?

A

Proteins, cytosol, and proteins compacted into different granules

29
Q

Stain with hematoxylin in…

A

Water

30
Q

Stain with eosin in…

A

Alcohol

31
Q

Does a Pancreas stained with H&E have polarity, how can you tell?

A

Red top half, contains a lot of proteins. Blue bottom half contains nucleus and the rough ER

32
Q

What does a brown color in stained skin cells represent?

A

Melanin

33
Q

Would hemoglobin show a nucleus in the center when stained?

A

No, hemoglobin is a mature RBC and do not have a nucleus

34
Q

Would blood vessels stain red or blue?

A

Red because RBC’s do not have a nucleus

35
Q

What technique is used when staining connective tissue?

A

Trichrome stain. Blue color shows cartilage, red shows muscle

36
Q

What staining technique is used for neurons?

A

Golgi (silver) stain

37
Q

What is the Feulgen reaction?

A

Treat tissue with HCl acid, purine base on DNA is cleaved, forming a aldehyde group which reacts with Schiff’s reagent

38
Q

What technique is used to stain Nucleic Acid?

A

Feulgen

39
Q

What is Periodic Acid Schiff (PAS) stain specific for?

A

Sugars

40
Q

What happens when glucose in a tissue sample is treated with periodic acid (HIO4)?

A

An aldehyde group is formed, which can react with schiff’s reagent

41
Q

What color is produced with an aldehyde group reacts with Schiffs reagent?

A

Red or magenta

42
Q

What staining technique would you use if you wanted to quantify the amount of sugar in the liver?

A

Periodic Acid Schiff (PAS) stain

43
Q

What technique would you use to visualize the basement membrane and brush border of kidney tubules?

A

Periodic Acid Schiff (PAS)

44
Q

Which staining technique is specific for fats?

A

Oil red O and Sudan Black

45
Q

Can you use paraffin embedded sections to visualize fat?

A

No, you actually remove fat from tissue using xylene when using the paraffin embedded tissue preparation

46
Q

Why do you need to use frozen sections to stain fats?

A

Fats are soluble in alcohol and xylene (paraffin embedded tissue). staining fat must be done in unfixed cells and tissue

47
Q

Structures that stain with eosin are called…

A

Acidophilic

48
Q

What is Immunocytochemistry/immunohistochemistry?

A

Antibody staining

49
Q

The antibody must be tagged. You can use…

A
  1. Enzymes
    Or
  2. Fluorescent molecules
    Or
  3. Colloidal gold
50
Q

Immunohistochemistry (IHC), uses what to tag antibodies?

A

Enzymes

51
Q

Immunoelectron microscopy

A

Antibodies are tagged with colloidal gold and viewed with an electron microscope

52
Q

Immunofluorescence

A

Antibodies are tagged with fluorescent molecules and viewed with a fluorescent microscope

53
Q

Immunoperoxidase stain (type of IHC)

A

Produces a brownish/black precipitate that is viewed on the slides.
Example: incubate pituitary cells with an antibody to growth hormone, and then a second antibody with the peroxidase enzyme on it. Peroxidase enzyme will detect the growth hormone, than add the DAB substrate, the peroxidase reacts with the peroxide, produces the brown product

54
Q

Which microscope has the higher resolution? Light or electron?

A

Electron microscope

55
Q

A transmission electron microscope is used to view…

A

Uses electrons that pass through tissue, identify objects that are inside a cell

56
Q

What would you use to visualize a 3D image of the surface of cells?

A

Scanning electron microscope (SEM), electrons are scattered

57
Q

Myelinated nerve axons must be viewed using…

A

Transmission electron microscope (TEM), inside a cell

58
Q

RBC in a fibrin clot must be viewed using a…

A

Scanning electron microscope (SEM)

59
Q

Good tip to identify Osteoclasts under the microscope?

A

Multiple nuclei

60
Q

What is the size of a Paraffin section diameter ?

A

Usually 5 to 7 micrometers

61
Q

RBC are found in most tissue, how big are RBC?

A

About 7 micrometers

62
Q

In paraffin embedded tissue preparation, which steps comes before dehydration with alcohols?

A

Fixing in a chemical such as formaldehyde

63
Q

Which microscope has the lowest resolution: fluorescent, scanning electron, or transmission electron?

A

Fluorescent

64
Q

In an H&E staining section, which dye stains basophilic structures and what feature of a cell would it stain?

A

Hematoxylin, it would stain the nucleus, RER, and mitochondria

65
Q

Why is mitochondria basophilic?

A

It contains DNA, which is negatively charged and binds to the positively charged hematoxylin dye

66
Q

Cytosol and secretory vesicles would stain [ ] due to positively charged proteins

A

Red, Eosin

Fundamentals Of Medicine: Exam 1 (18 decks)

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