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Stain principles Flashcards

1
Q

What is the most common tissue preservation technique?

A

Fixing in 10% Formalin (4% formaldehyde)

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2
Q

What percentage of tissues received in Histopathology are fixed in 10% formalin?

A

99%

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3
Q

What is the typical thickness for sections cut by microtomy?

A

4 or 5 microns (5 μm)

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4
Q

What happens to haematoxylin during the blueing process?

A

It converts from pink to blue as pH raises.

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5
Q

What is the purpose of differentiation in 1% acid-alcohol?

A

Selective removal of excess dye from weakly bound components

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6
Q

What are mordants?

A

Substances/metal which chemically binds with dye and tissue

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7
Q

What does haematoxylin oxidation produce?

A

Haematin

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8
Q

What is the purpose of dehydration in tissue processing?

A

To prepare sections for embedding in organic solvents

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9
Q

What is the purpose of dewaxing and rehydrating tissue sections?

A

Removes wax and rehydrates tissues for aqueous staining

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10
Q

What is the significance of adhesive or charged slides?

A

They attract sections to the glass slide

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11
Q

What is Cryotomy?

A

Use of refrigerated microtome to cut frozen tissue sections

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12
Q

What is the commonest lipid stain used?

A

Oil red O

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13
Q

What is the main application of rapid Haematoxylin and Eosin staining?

A

Urgent diagnosis during surgery

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14
Q

What does H&E staining specifically stain?

A

Nuclei blue (Haematoxylin) and cytoplasm pink (Eosin)

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15
Q

What is the role of Eosin in H&E staining?

A

Counterstain that contrasts with Haematoxylin

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16
Q

What are the criteria for a good H&E stain?

A
  • Chromatin detail
  • Colour
  • Differentiation
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17
Q

What is the principle of Trichrome staining?

A

Differentiates between tissue components, highlighting collagen and muscle

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18
Q

How do trichrome stains work?

A

Based on dye permeability, molecular size, and tissue affinity

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19
Q

What is the Masson trichrome stain primarily used for?

A

Highlighting collagen fibres, muscle fibres, and cytoplasm

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20
Q

What components are involved in Verhoeffs Van Gieson staining?

A
  • Verhoeff stain for elastic fibers
  • Van Gieson stain for collagen
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21
Q

What is the function of Gordan and Sweet’s method?

A

Demonstration of argyrophilic reticulin fibres

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22
Q

What does Perls Prussian Blue stain detect?

A

Ferric (Fe3+) iron deposits

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23
Q

What is the principle of Alcian Blue/Periodic Acid-Schiff stain?

A

Determines presence of carbohydrates and classifies them

24
Q

What unique property does Congo Red stain have?

A

Binds to amyloid proteins and exhibits apple-green birefringence under polarized light

25
What is the role of Grocott’s silver stain?
Detects fungal cell walls rich in polysaccharides
26
What is the ABC method in immunohistochemistry?
A three-step antibody detection method
27
What is the function of PBS in immunohistochemistry?
Washing buffer to remove excess antibodies and stabilize bindings
28
What is the Cytospin method used for?
Produces a monolayer of cells for screening
29
What is visualized as a brown deposit in antigen-antibody reactions?
The antigen-antibody reaction ## Footnote This indicates the presence of the specific antibody bound to the antigen.
30
What is the purpose of Dakopen in immunohistochemistry?
Confines liquid inside due to its hydrophobic nature ## Footnote This property helps in the proper visualization of the antigen-antibody reaction.
31
What are chromogens used for in immunohistochemistry?
Produce a coloured product that can be visualized by light microscopy ## Footnote They react with specific antibodies to provide contrast in staining.
32
What is the function of PBS in immunohistochemistry?
Used as a washing buffer to remove excess antibodies and reagents ## Footnote It also stabilizes antigen-antibody binding.
33
What is the Cytospin method used for in clinical cytology?
Produces a monolayer of cells in a small area for screening ## Footnote It is particularly useful in laboratories with large workloads.
34
What types of specimens can be processed using the Cytospin method?
Both fixed and unfixed specimens ## Footnote Unfixed samples must be post-fixed before staining.
35
What is crucial in obtaining a monolayer using the Cytospin method?
The concentration of the re-suspended cell pellet ## Footnote Proper concentration ensures even distribution of cells.
36
What speed and duration are used in a cytocentrifuge for the Cytospin method?
1500 rpm for 5 minutes ## Footnote Followed by fixation in spirit for 5 minutes.
37
What is the main advantage of direct smears in clinical cytology?
Quick and simple method to prepare slides for screening ## Footnote It is primarily employed for Diagnostic Cytology.
38
What is used to represent a diagnostic cytology specimen in direct smears?
Cultured cell lines ## Footnote This allows for easier analysis and identification of cellular characteristics.
39
What is the process of preparing a direct slide from a centrifuged specimen?
Specimens are centrifuged, supernatant decanted, and cell pellet agitated to prepare a sample ## Footnote This ensures a concentrated and even cell distribution on the slide.
40
What technology is utilized in liquid-based cytology for cervical cytology?
ThinPrep technology ## Footnote It is also applicable for diagnostic cytology.
41
What is the basis of the ThinPrep process in liquid-based cytology?
Filtration method ## Footnote This method enhances the quality of the cell monolayer on slides.
42
What pre-treatment is used for cell samples in liquid-based cytology?
Cytolyt ## Footnote This is done prior to transferring to PreservCyt vials.
43
What is the fixation medium used in liquid-based cytology?
PreservCyt ## Footnote It preserves the cell samples before processing.
44
What does the Papanicolaou stain principle require for immediate fixation?
95% alcohol ## Footnote This prevents air drying of the samples.
45
What type of dye is Harris Haematoxylin?
Basic dye that binds to acidic sulfate groups of DNA ## Footnote It is essential for the staining of cellular structures.
46
What is the purpose of differentiation in Papanicolaou staining?
To selectively remove excess stain from all except the structures to be demonstrated ## Footnote This enhances the visibility of the target cells.
47
What happens during the blueing process in Papanicolaou staining?
Alum haematoxylin stain converts red colour to blue-black ## Footnote This occurs when washed in a weak alkali solution.
48
What are the colours associated with different cell types in Papanicolaou staining?
Nuclei -> blue/black; Cytoplasm -> spectrum of pink, orange, green ## Footnote Specific colours indicate different cellular components and types.
49
What is the sensitivity and accuracy of the Papanicolaou test?
Low sensitivity with limited accuracy ## Footnote It is primarily a screening test.
50
What is the Speedy-Diff stain principle?
Commercial Romanowsky stain used to rapidly stain and differentiate pathology specimens ## Footnote It can be applied to air-dried samples.
51
What colour do nuclei appear in Speedy-Diff staining?
Deep blue/purple ## Footnote This contrasts with the cytoplasm's blue to mauve colour.
52
What is the colour of RBCs in Speedy-Diff staining?
Salmon pink ## Footnote This indicates the presence of red blood cells in the sample.
53
Decalcification
Some tissues contain calcium deposits which are extremely firm and will not section properly with paraffin embedding. Typically bone specimens. Calcium is removed using mineral acids such as HCL (can cause cellular damage) or using EDTA (slow + expensive).
54
Presence of fine black precipitate on slides
Suggests formaline-heme pigment formation. Occurs when tissue become acidic and complex forms.
55
Tissue insufficiently dehydrate prior to clearing and infiltration with paraffin wax
Hard to section on microtome and tearing artefacts + holes will occur in sections.
56
Alcohol as fixative - artefacts
Can make tissue sections brittle resulting in microtome sectioning artefacts such as chattering.

Cell Pathology (28 decks)

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