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Transfusion > Cell path practicals > Flashcards

Cell path practicals Flashcards

1
Q

what is the purpose of differentiation

A

removing excess stain - we regressively stain with haematoxylin (over stain) to remove later
(removing background)

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2
Q

why do sections need bluing after haematoxylin staining

A

ammonia (bluing solution) changes the red-purple haem to a blue purple stain so is more distinguishable and clear to identify from the eosin colours

The blueing solution also has an alkaline pH. This re forms a mordant-dye lake in the tissue which makes the blue dye more permanent and insoluble compared to the purple one

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3
Q

what conclusions are drawn from H&E staining

A

staining of normal tissue sections
Haem - basic dye which stains DNA -
chromosomes- deep blue

Eosin- acidic dye which stains proteins -
muscle- deep pink
collagen- pale pink
RBC- orange

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4
Q

What cell type contains the iron stain

A

macrophages

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5
Q

what is the principle of PERLs staining

A
  1. HCL splits off the bounded proteins allowing potassium Ferro cyanide to combine with Ferric ions producing a Prussian Blue colour.
  2. Staining time vary depending on the amount of ferrous ions present.
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6
Q

what are tissue deposits containing ferric ions called

A

hemosiderin deposits?

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7
Q

what is the difference between haemosiderosis and haemochromatosis

A

haemosiderosis - focal deposition of iron - no tissue damage

haemochromatosis - iron overload - typically systematic process in which iron deposits cause tissue damage

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8
Q

what is the purpose of perls Prussian blue

A

demonstrate ferric ions location

collagen - red
muscle - yellow
nuclei - red

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9
Q

what counterstain is used in perls Prussian blue

A

van gieson - collagen - red
muscle - yellow
neutal red - nuclei - red

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10
Q

Perls: what tissue is used for positive control?

A

Sections containing Ferric ions e.g Spleen, Liver, Bone Marrow

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11
Q

What are Perls Reagents?

A

HCL acid and Potassium Ferrocyanide

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12
Q

explain the function of HCL in Perls Stain

A

HCL splits off the bounded proteins allowing to stain blue

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13
Q

What is the purpose of a Pigment Special Stain - Formalin Pigment?

A

to compare the presence of formalin pigment (appearing as dark brown, black granules) around red blood cell

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14
Q

why does formalin pigmentation occur?

A
  • deposits formed by the action of acidic (pH<5.6) fixatives on Haemoglobin.
  • tissues stored for long times
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15
Q

What is the principle of Formalin Pigment Removal?

A

Alcoholic Picric Acid is used to remove formalin pigment then stained using routine H&E to see if the pigment is still present.

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16
Q

what does PAS stain

A 
hepatocytes - glycogen
mucins
mucoprotein
glycoprotein
fungi
17
Q

what is the counterstain in PAS

A

haematoxylin

18
Q

what is the principle of PAS staining

A

exposes tissue to peroic acid which oxidises compounds with free OH groups, resulting in dialdehydes = Schiffs create magenta coloured complex formation.

19
Q

what type of cells contain the lipofuscin pigment

A

liver, kidney, heart muscle, retina, adrenals, nerve cells, ganglion cells

20
Q

where in the cell is the lipofuscin located

A

in cytoplasm around the nucleus

21
Q

what cells contain lipochromes

A

aging post mitotic cells

22
Q

what other histochemical stains identify Lipofuscin apart from PAS

A

sudan black B

long Ziehl-neelson acid fast

23
Q

what is lipofuscin also known as

A

the wear and tear pigment

24
Q

how do cells produce lipofuscin

A

byproduct of failed intracellular catabolism

25
Q

what is it believed about excessive lipofuscin?

A

causes cell death - not just a bystander

26
Q

what colours and their organelles are shown in PAS staining

A

PAS - red/magenta - glycogen, basement membranes, reticular fibres, cartilage. or superficial fungal elements if candida infection

27
Q

what does PAS demonstrate

A

carbohydrates in tissues (glycogen)

28
Q

basic principles of PAS staining?

A

periodic acid oxidises tissue carbs, producing aldehyde groups, Schiffs reagent condenses these an forms bright red colouration.

29
Q

conditions necessary for a PAS positive reaction

A
  1. must contain the 1-2 glycol grouping, an equivalent amino or alkylamino grouping, or the oxidation product CHOH-CO
  2. must not diffuse away after fixation
  3. must give an oxidation product that doesn’t diffuse away
  4. must be present in sufficient concentration to produce detectable final colour

Transfusion (6 decks)

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