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Microscopic Anatomy > Part 1: Introduction and Cell Components > Flashcards

Part 1: Introduction and Cell Components Flashcards

1
Q

how does electron microscopy work

A

it uses a beam of accelerated electrons as a source of illumination

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2
Q

what type of electron microscope creates 3-D but may require special preparation of the sample

A

scanning electron microscope

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3
Q

what type of electron microscope offers the greatest magnification, high resolution in two dimensions, and uses a fluorescent screen

A

transmission electron microscope

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4
Q

what is the relationship between speed of an electron and wavelength of an electron

A

there is a direct correlation

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5
Q

what is the % of formaldehyde in aqeuous buffered formalin

A

37%

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6
Q

what is the pH of formalin

A

7.0 - neutral

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7
Q

what are four examples of formed elements seen in tissue after fixation

A

nucleoproteins
intracellular cytoskeletal proteins
extracellular proteins
membrane phospholipid-protein or carb complexes

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8
Q

what substance clears tissue of alcohol before it is imbedded into paraffin

A

xylene

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9
Q

how thick are usual histologic sections

A

typically 4 microns

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10
Q

what is the common mounting media used to coverslip tissue

A

paramount

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11
Q

what is the first stain used in the staining process

A

hematoxylin

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12
Q

what counter stain is used in the staining process

A

eosin

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13
Q

what salt additive is used as a mordant to make hematoxylin act as a basic dye

A

aluminum

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14
Q

eosin attaches to what charged components in a cell

A

catatonic, + charged

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15
Q

what is a heavy metal that can be used to bind with lipids in the tissue commonly used in EM

A

osmium tetroxide

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16
Q

what is a good preservative to use when submitted tissue for EM

A

glutaraldehyde

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17
Q

what is a common transport media for live cells, especially lymph nodes

A

RPMI

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18
Q

what temperature is used for a cryostat

A

-15 to -20 degrees celsius

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19
Q

what methods of snap freezing causes the least amount of damage

A

isopentane

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20
Q

what is the most common method of snap freezing

A

liquid nitrogen (-196 degrees celsius)

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21
Q

what staining technique may be used to identify lipids during a frozen section

A

thionine

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22
Q

what stain is used for a parathyroid frozen

A

oil red-o

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23
Q

what is the name of the part of the antibody that binds to the antigen in IHC

A

epitope

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24
Q

what are the common reagents used for blocking in IHC

A

3% of both peroxidase and serum albumin

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25
Q

what are the three methods of labeling antibodies in IHC

A

flurochromes
enzymes
EM

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26
Q

cytokeratin identifies what

A

carcinoma and adenocarcinoma

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27
Q

vimentin identifies what

A

CT tumors and melanoma

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28
Q

LCA identifies what

A

leukemia and lymphoma

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29
Q

CD20 identifies what

A

B lymphocytes

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30
Q

CD3 identifies what

A

T lymphocytes

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31
Q

CD34 identifies what

A

angiomas, angiosarcoma, and fibroblastic tumors

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32
Q

SMA identifes what

A

myofibroblastic and myoepithelial tumors

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33
Q

desmin identifies what

A

muscle

34
Q

S-100 identifies what

A

neutral cells, cartilage, granular cell tumors, and melanomas

35
Q

HMB-45 identifes what

A

some melanomas

36
Q

what is the name for the study of tissue structure

A

microanatomy

37
Q

what are the 5 ways electrons react to an object in electron microscopy

A

unscattered (go right through)
secondary electrons
auger electrons
form an x-ray
back scattered

38
Q

what are some disadvantages of electron microscopy

A

expensive
large machine
sensitive to things like vibrations
specimen must be in a vacuum chamber
needs a constant voltage supply
needs circulation of cool water
needs personnel training

39
Q

what are some advantages of anatomic force microscope (a type better than electron microscopy)

A

sample preperation is easier
works in a vacuum, air, and in liquids
living systems can be studied, no fixation needed
higher resolution then TEM

40
Q

what are some disadvantages of anatomic force microscope

A

limited vertical range
limited magnification range
tip or sample can be damaged

41
Q

why is super resolution microscopy so affective

A

allows for visualization of subcellular organization with lots of details

42
Q

what are the three main steps associated with routine tissue preparation

A

formalin fixation
paraffin embedding and cutting
staining

43
Q

what properties does formalin fixation provide

A

terminate cell metabolism
prevent enzymatic degradation of cells by autolysis
kills pathogens
hardens tissue
maintains general structure of cell

44
Q

what process must occur to tissue on a slide before it can be stained

A

dissolve paraffin with xylene
rehydrate tissue using decreasing alcohols

45
Q

how is water removed after washing the formalin from the tissue

A

using a series of dehydrating alcohol

46
Q

why must the tissue go through a series of alcohols before staining with eosin

A

eosin is more soluble in alcohol than water

47
Q

which stain will show nuclear DNA and cytoplasmic RNA more

A

hematoxylin

48
Q

hematoxylin attaches to what charged components in a cell

A

anionic, - charged

49
Q

what is an example of a negatively charged component (basophilic) in a cell

A

nucleus

50
Q

what is an example of positively charged component (acidophilic) in a cell

A

proteins in the cytoplasm

51
Q

what does basophilic mean in tissue staining

A

tissue readily stains with basic dyes such as hematoxylin

52
Q

what does acidophilic mean in tissue staining

A

tissue readily stains with acid dyes such as eosin

53
Q

what is a good preservative when submitting skin for immunofluorescence

A

Zeus (Michele’s media)

54
Q

why would you do frozen section on tissue

A

no preoperative diagnosis
unexpected intraoperative finding
evaluation of surgical margins

55
Q

what media is used in performing a frozen section

A

OCT (optimum cutting temperature)

56
Q

what are the three steps to prepare a frozen section

A

freeze tissue in OCT
cut tissue in cryostat
stain tissue on a slide and cover slip

57
Q

immunohistochemistry targets antigens within the tissue by applying what substance

A

antibodies tagged with visual markers

58
Q

what are some uses of immunohistochemistry

A

identify replicating cells, signaling cells, apoptotic cells, activation states, and different types of cells within the same tissue

59
Q

what are the benefits of formalin fixation in IHC

A

antibodies preform optimally
prevents degradation of tissue
preserves position of antigen

60
Q

what is the advantage of freezing tissue in liquid nitrogen or isopentane for IHC

A

the antigen is preserved

61
Q

what is the disadvantage of freezing tissue in liquid nitrogen or isopentane for IHC

A

freezing artifact alters cellular morphology

62
Q

what are two methods of antigen retrieval for IHC

A

HIER (heat inducted epitope retrieval)
enzyme digestion

63
Q

what is the main purpose of the antigen retrieval process in IHC

A

to increase the accessibility of the antibody to the antigen

64
Q

what is the purpose of permeabilization in IHC

A

to improve antibody penetration to access nuclear and cytoplasmic antigens

65
Q

what is the purpose of blocking in IHC

A

to eliminate background staining

66
Q

what are some disadvantages of using direct IHC techniques

A

less signal amplification so may not get a signal with little antigen
conjugation process may interfere with the antibody to react with target antigen

67
Q

what is the main advantage of indirect IHC

A

sensitivity of antibody reacting to the target antigen is increased because multiple secondary antibodies can bind to a single primary antibody amplifying the signal

68
Q

what are the main differences between monoclonal and polyclonal antibodies used in IHC testing

A

monoclonal are consistent from batch to batch but may have false negative results
polyclonal tend to have non-specific reactivity and vary from batch to batch

69
Q

how are the results of IHC testing validated

A

using known positive and negative controls

70
Q

what are some advantages of enzymatic methods of IHC

A

they can be viewed through a standard light microscope
enzyme table specimens have an unlimited shelf life

71
Q

what are some disadvantages of enzymatic methods as compared to fluorescent methods in IHC

A

fluorescence has better resolution
enzyme intubation process causes testing to take longer
enzyme amplification interferes with signal quantization
substrates used to activate enzymes can be toxic or carcinogenic

72
Q

positive controls are useful in IHC in detecting false negatives caused by what

A

poor fixation of tissue
problem with tissue processing

73
Q

what are negative controls in IHC useful in detecting

A

endogenous biotin
peroxidase activity

74
Q

what information does IHC provide a Pathologist

A

rendering a diagnosis when morphology alone isn’t enough
differentiation of a tumor
often used as prognosis and predictive markers in therapeutic treatment

75
Q

what microbiology technique is preformed on cultured cells or embryos to localize specific nuclear sequences as small as 10-20 copies of mRNA or DNA

A

in situ hybridization

76
Q

what microbiology technique is used to detect minute quantities of rare or single copy number nucleic acids sequences in frozen or embedded cells or tissue sections for the localization of those sequences within the cells

A

PCR

77
Q

what is a single-stranded nucleic acid employed in the hybridization, requiring a color molecule to attach to it

A

a probe

78
Q

what is used in the amplification of a sample in a hybridization technique that is significantly shorter than a probe

A

a primer

79
Q

what type of test is used to determine trace amount of DNA in a sample

A

PCR

80
Q

what type of test is appropriate for determining if there are antibodies present in the blood as a result of exposure

A

serology testing

Microscopic Anatomy (11 decks)

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