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Histology - Overview and Staining methods Flashcards

1
Q

what are the 4 main building blocks of life

A
  • proteins
  • lipids
  • nucleic acid
  • carbohydrates
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2
Q

what is histology

A

a microscopic study of normal cells and tissues, looking at how structure is related to tissue function, and how disease is associated with disruption of normal cell structure and function

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3
Q

what is pathology

A

a microscopic study of diseased cells and tissues

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4
Q

what are the 4 main tissue types

A

epithelial
connective
muscle
nervous

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5
Q

what are the two main methods of microscopy

A

light microscopy and electron microscopy

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6
Q

what is light microscopy

A

low resolution microscopy used to view basic cell structure

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7
Q

what is electron microscopy

A

method for viewing the ultrastructure of the cell

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8
Q

what is the microscopy technique used in routine histology

A

light microscopy

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9
Q

what are the steps to microscopy

A
  • specimen collection
  • fixation
  • dehydration
  • embedding
  • sectioning
  • staining
  • viewing
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10
Q

why are tissue samples cut into thin sections prior to the microscopy

A

most tissue samples are too thick to be examined directly under a microscope and must first be cut into very thin sections to enable light or electrons to pass through the tissue and make the structure visible

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11
Q

what are the different technique to specimen collection

A
  • incision or a punch biopsy
  • needle biopsy
  • endoscopic biopsy
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12
Q

what are incision or punch biospies used for

A

specimen collection from skin or oral surfaces

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13
Q

what are needle biopsies used for

A

specimen collection from organs or lumps below the skin. to guide the biopsy, imaging techniques like x rays and mris can be used

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14
Q

what does endoscopic biopsy involve

A

a flexible tube with light and a camera - cutting tools can be used to collect tissue specimens

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15
Q

where would an endoscopic biopsy be used in the body

A
  • trachea
  • bronchus
  • lungs
  • esophagus
  • stomach
  • small intestine
  • colon and rectum
  • mainly bladder
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16
Q

where would a needle biopsy be used

A

the brain
eye
thyroid
lymph nodes
breast
lung and pleura
liver
kidney
bone and bone marrow
testis
skeletal muscle

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17
Q

what kind of biopsy is used for the heart and the liver

A

transvascular biopsy

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18
Q

what do cells release as they die

A

enzymes that degrade components for the tissue

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19
Q

what does fixation do

A

terminates all biochemical reactions and so prevents the release and activity of degradative enzymes as well as inhibiting microbial growth and so prevents tissue decomposition

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20
Q

what are some common fixatives

A

formaldehyde and glutaraldehyde

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21
Q

why is fixation important during microscopy

A

it preserves the structural arrangement betwen cells and extracellular components

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22
Q

is paraffin embedding compatible with water

A

no

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23
Q

how is water removed from tissues during dehydration

A

using a series of graded alcohols, done gradually to be prevent distortion

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24
Q

why should water not be removed from the tissue sample in a single step

A

the tissue would shrink rapidly and the structural arrangement would be distorted

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25
why do tissues require embedding
thin tissue sections are delicate and soft and easily distorted, and must be embedded in a hard substance as support
26
what are most commonly used to embed tissue samples
resin and paraffin
27
what are tissue samples submersed in prior to embedding and why
resin and paraffin are not compatible with water or alcohol, so after dehydration the sample is then immersed for a period of time in a strong solvent like xylene to replace the alcohol
28
how should tissue be cut to be viewed under a microscope
thin and flat, ideally just a single layer of cells
29
why should tissue samples be cut into thin and flat sections
so they are transparent, as this allows the beam of light or electrons to pass through the tissue to enable visualisation
30
how are embedded speciments sectioned
using a microtome
31
what is a microtome
a machine that cuts embedded tissues into sections around 7 nanometres thick
32
what are tissue sections mounted on
glass slides
33
why are tissue sections mounted on glass slides
for protection
34
why is staining another important step in microscopy
most cells are colourless and transparent, so staining makes cells and their components more visible
35
is there a limited number of staining methods available for light microscopy
no there is a wide variety
36
what does staining allow
identification of different cell and tissue features
37
what state are most stains for light microscopy
aqueous
38
are stains compatible with paraffin
no
39
why are stains not compatible with paraffin
they are aqueous
40
how do you use stains incompatible with paraffin
reversal of the dehydration steps used earlier, so the embedded sample is first submerged in xylene before then being passed through the graded series of alcohol in order to replace the alcohol with water and leave it ready for staining
41
does where the tissues are sectioned make much of a difference to tissue architecture
yes
42
what is the most commonly used staining technique
haematoxylin and eosin (H&E)
43
what is haematoxylin
a basic dye that stains acidic structures like nucleic acid blue/purple
44
what is eosin
an acidic dye that stains basic structures such as cytoplasmic proteins red or pink
45
are epithelial tissues highly cellular
yes - there is very little extracellular material
46
what is the periodic acid-schiff reaction (PAS)
a method that stains complex carbohydrates magenta
47
what are mucins produced by
goblet cells
48
what are examples of PAS-positive molecules
muines produced by goblet cells, brush borders and basement membranes
49
what is the proximal convoluted tubule
part of the nephron in the kidney. involved in reabsorption of essential nutrients and water to prevent them being excreted within urine
50
why does the proximated convoluted tubule require a large surface area
it is involved in reabsorption of essential nutrients and water
51
what does the PAS staining method reveal in the proximated convulated tubule
the prominent brush border projecting into the lumen of the tubule, as well as the basement membrane, as both are strongly PAS positive.
52
what is the oesophagus
a strong muscular tube that conveys food from the throat to the stomach
53
how is the mucosa found when in a relaxed state
deeply folded
54
what do the different colours of masson trichrome represent
the blue purple colour is the nuclei, and other basophilic structures the green blue is collagen the red is the cytoplasm, muscle, red blood cells and keratin
55
explain the immunohisotochemistry technique for staining
utilises antibody specifity for antigen. antibodies are required to identify substances of interest. the antibody is conjugated to an indicator, whether enzymatic or fluorescecnt
56
what is the most specific staining method
immunohistochemistry
57
what does immunohistochemistry use antibodies to do
stain specific cellular proteins by using antibodies specific for their antigen to reveal the components by joining the antibody to some sort of colour indicator
58
what is the alcian blue staining method
- stains mucin and cartilage blue, and can be combined with other stains like H&E
59
what is the van gieson staining technique
stains collagen red, nuclei and erythrocytes and cytoplasm yellow
60
what are reticulin fibres components of
connective tissue
61
what is the reticulin stain
this stains reticulin fibres blue black, and is often combined with H&E
62
what is the most common form of methods used in histology
light microscopy
63
how can light microscopy be subdivided
bright field, immunofluorescence and dark field microscopy
64
give a breakdown of periodic acid schiff
this is a method that stains complex carbohydrates a dark red or magenta. it identifies glycogen in cells
65
what are the main steps to preparing tissue for bright field microscopy
- fix the tissue using formalin - process the tissue (remove water with ethanol, remove ethanol with xylene. embed in paraffin - removes xylene) - section
66
what happens once the paraffin embedded tissue samples are sliced accordingly
they are mounted on glass slides. staining makes it easier to then see under the microscope and identify the cell components
67
what is the most common staining method used in bright field microscopy
haemotoxylin and eosin (H&E)
68
give a breakdown on how H&E staining works
the haemotoxylin is positively charged and stains basophilic structures, which are negatively charged. these include nuclei, ribosomes, cartilage matrix and the rough endoplasmic reticulum the eosin is negatively charged and stains eosinophilic structures, which are positively charged, pink. these include collagen, cytoplasmic proteins and mitochondria
69
how does the giemsa stain technique work
this is the standard stain for blood and bone marrow smears
70
how does alcian blue stain work
this stains acidic polysaccharides blue, like mucin and cartilage
71
what does van gieson stain
blood vessels and the skin
72
explain the different staining colours of masson trichrome
stains connective tissue blue stains nuclei dark red or purple stains cytoplasm, kertain, and erythro red or pink
73
what does masson trichrome stain
supporting tissues like collagen
74
what is PAS
periodic acid schiff
75
76
what is the masson trichrome staining technique used for
staining connective tissue - useful for differentiating different types of tissues

Histology (10 decks)

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