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pathology coursework exam > practical 1- H&E staining > Flashcards

practical 1- H&E staining Flashcards

1
Q

what governs the overall staining pattern

A

rate of which a dye is taken up by a tissue component or the rate of loss of due from a tissue component

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2
Q

what are the 3 layers composing the skin

A

epidermis dermis subcutis (hypodermis)

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3
Q

In H&E staining, how do we differentiate a section after staining with harris’s haematoxylin and why do we do this

A

differentiate in 1% acid alcohol for 2-5 seconds use of acidic solutions to remove excess background staining

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4
Q

In H&E staining, what step is taken after dewaxing, hydrating, haematoxylin staining, differentiating and blueing

A

immerse in eosin for 2-4 minutes

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5
Q

in a rate depending staining process, what is a progressive dye

A

dyes staining more intensely the longer the tissue remains in the dye bath

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6
Q

what colour stain does eosin produce

A

red cytoplasmic stain

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7
Q

where might you see cells in the process of dividing (mitosis)

A

mucosa- base of intestinal crypts, this is to replenish cells at the epithelium

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8
Q

In H&E staining, what blueing reagent is used and why do we use it?

A

Scotts solution necessary to convert nuclear colouration from reddish purple to crisp blue/purple enhances the contrast of the H&E stain by increasing the crispness of the haematoxylin

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9
Q

what is meant by basophilia

A

tissues with affinity for haematoxylin (basic dye) is base loving

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10
Q

what is the function of the muscularis externa

A

undergoes segmentation contractions (which mixes chyme in the small intestine) and weak peristaltic contraction (moves chyme along in the small intestine)

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11
Q

what does harris’s haematoxylin use to alter the specificity of its stain explain the procedure

A

uses aluminium salts (haemalum) as a mordant rinse slide in tap water to blue the section haematoxylin used regressively with controlled differentiation in acid alcohol reblueing in tap water (must be alkaline)

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12
Q

how many layers of cells are there in the absorptive epithelium proper

A

mucosa- one layer

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13
Q

what is the function of the submucosa

A

contains brunners galnds, they secrete alkaline mucus which neutralises gastric acid in chyme

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14
Q

in a rate depending staining process, what is a regressive dye

A

a dye may be selectively lost/removed fro a tissue to distiniguish/differentiate between components with stronger or weaker affinities for the dye

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15
Q

how is haematoxylin produced

A

extracted in water and precipitated with urea

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16
Q

what type of dye are the following Ehrilchs, Mayers, Harris, Gills, Coles, Delafield, Carazzis

A

haematoxylin dyes

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17
Q

what type of a staining procedure is H&E staining

A

regressive

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18
Q

what is the function of enteroendocrine cells and paneth cells

A

enteroendocrine cells secrete hormones (somatostatin) paneth cells have an immune cell function

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19
Q

what is eosin and how does it work

A

acidic dye, but binds to basic (eosinophillic) compounds/structures

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20
Q

see the skin diagram in the test booklet, label A-L

A

A- stratium corneum (cornified/horny layer) B- pickle cell layer C- dermis D- fat E- sweat gland F- hair follicle G- blood vessels H- sebaceous (oil) gland I- lymph vessel J- sensory nerve receptor K- hair L- pores

21
Q

In H&E staining, how do we dewax a section

A

place in histoclear for 5 minutes

22
Q

what is meant by acidophilia

A

tissues with affinity for eosin (acidic dye) is acid loving

23
Q

look at tissue A in the test booklet, label A, B and C how are the columns of hepatocytes that radiate from the central vein seperated

A

A- portal triad (proper hepatic artery, hepatic portal vein and bile duct) B- hepatic vein C- hexagonal hepatic lobule (bile canaliculi, sinusoidal capillaries, hepatocytes) seperated by canniculae of the bile ducts

24
Q

In H&E staining, what do we do after dewaxing and hydrating a sample

A

immerse in Harris’s haematoxylin for 5 minutes

25
Q

what alters specificity of a haematoxylin stain

A

use of a mordant (inorganic stain-fixing solution)

26
Q

what is the most common method used to enhance contrast between cell structures and tissues

A

dyes e.g. Haematoxylin and Eosin staining

27
Q

where does haematoxylin come from

A

natural product made from haematoxylum campechianum (logwood)

28
Q

what are the 3 zones in the duodenum state what each zone consists of

A

mucosa- inner absorptive epithelium with associated connective tissue (such as blood vessels) consists of lamina propria and muscularis mucosa submucosa- underlying zone of loos CT muscularis externa- outer muscle coat, consists of circular and longitudinal layers

29
Q

what is meant by eosinophilia

A

eosin loving, dye term used to describe tissue affinity

30
Q

what colour staining does H&E produce

A

red/blue

31
Q

what procedure does H&E staining rely on

A

acid-base dye interactions

32
Q

what is the function and structure of the subcutis (hypodermis)

A

layer containing main vascular supply contains adipose tissue and fibrous tissue

33
Q

what is the function and structure of the epidermis name the layers, inner to outer

A

upper, protective zone comprising several sublayers of stratified squamous epithelium boys smell girls like chocolate basale, spinosum, granulosum, corneum

34
Q

what are the different types of eosin stains

A

eosin Y and eosin B

35
Q

In H&E staining, how do we dehydrate a sample

A

by passing through alcohols of increasing concentration 70% alcohol for 1 minute 90% alcohol for 1 minute absolution (100%) alcohol for 1 minute

36
Q

what is the function and structure of the dermis

A

underlying zone containing several glands and structures which include: hair follicles, sebaceous glands, eccrine sweat glands and apocrine glands

37
Q

look at the diagram of tissue C- duodenum on the test booklet and label A-G

A

A- epithelial layer of cells B- villus C- lamina propria D- muscularis mucosae (mucosa) E- intestinal crypts (crypt of lieberkuhn) F- brunners gland G- serosa (outer layer)

38
Q

how do goblet cells and enterocytes differ

A

goblet cells are cup shaped (involution doesnt stain)- secrete mucus enterocytes are rectangular and have an elongated nuclei

39
Q

what is meant by dye-tissue affinity in H&E staining

A

measure of a dye to transfer from a dye bath onto a tissue section

40
Q

how is the muscularis externa arranged

A

outer thinner layer has longitudinal arrangement inner thicker layer has circular arrangement

41
Q

put the following duodenum layers in order (inner to outer) lamina propria sub mucosa adventitia epithelium muscularis propria muscularis mucosa

A

epithelium lamina propria muscularis mucosa sub mucosa muscularis propria adventitia

42
Q

name 3 other methods used to enhance contrast between cell structures and tissues

A

polarised light, dark field, phase contrast

43
Q

how thin must tissue sections be when viewing through a light microscope

A

<15µm

44
Q

what process must a haematoxylin dye undergo before use as a stain

A

requires oxidation to haematein

45
Q

In H&E staining, how is the sample cleared at the end

A

immersed in histoclear for 5 minutes and mounted in DePeX

46
Q

what does a haematoxylin dye stain, give examples

A

basophillic structures such as nucleic acids, ribosomes

47
Q

In H&E staining, how do we hydrate a sample

A

by passing through alcohols of decreasing concentration 100% (absolute) alcohol for 1 minute 90% alcohol for 1 minute 70% alcohol for 1 minute

48
Q

what is eosin derived from

A

fluorescein (fluoresces)

pathology coursework exam (5 decks)

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