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2.2 Cellular Pathology > Cytology Specimen Processing > Flashcards

Cytology Specimen Processing Flashcards

1
Q

What does cervical cytology involve?
(3)

A

Screening for cervical cancer

HPV screening

Cytology triage

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2
Q

What does diagnostic cytology invovle
(3)

A

Not screening

First line investigation

Sensitive and accurate
- ancillary techniques

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3
Q

What are the two main cervical cytology samples

A

PreservCyt vial
Buffered methanol

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4
Q

What are the six main diagnostic cytology samples

A

Joint fluid
CSF
Urine
Body cavity fluids
Respiratory samples
FNA samples

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5
Q

What body cavity fluids can be seen in diagnostic cytology samples?

A

Pleural
Peritoneal
Acetic fluid

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6
Q

What Respiratory samples can be seen in diagnostic cytology samples?
(4)

A

Sputum
Bronchial wash
Bronchial brush
Bronchoalveolar lavage

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7
Q

What FNA samples can be seen in the diagnostic cytology lab

A

Breast
Thyroid
Lymph node
Other organs

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8
Q

Why might a Urine sample be sent to diagnostic cytology lab

A

Looking for bladder cancer

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9
Q

Why might a CSF sample be sent to diagnostic cytology lab

A

Looking for lymphocytes, metastatic diseases or lymphomas

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10
Q

Why might joint fluid be sent to diagnostic cytology lab

A

Looking for ctystals -> goitre and pseudo goitre

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11
Q

How is cervical cytology carried out
(4)

A

Smear test put in LBC medium

Vial received in lab at specimen reception

HPV testing carried out

If HPV positive needs to be processed, screened and reported

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12
Q

Explain what happens if a smear test comes up HPV positive
(3)

A

Processed for cytology screening: Slide prep + pap stain

Screening: Medical scientists

Reporting: Pathologist

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13
Q

How do we process our slides in cytology

A

Smear test

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14
Q

How do we remove blood from smears

A

Glacial acetic acid

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15
Q

What do we do if we see dyskaryosis on a thin prep

A

Sent the woman on for colposcopy

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16
Q

How do we screen for HPV

A

Screen for HPV DNA

Roche Cobas 4800 assay - DNA based

Hologic Aptima HPV test - RNA based

Hologic - Genius (AI digital cytopathology)

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17
Q

What does the Hologic - Genius (AI digital cytopathology) do

A

This allows us to view cells on a computer screen -> don’t have to set up a microscope

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18
Q

What do we look for on cervical cytology
(4)

A

Triage test following HPV positive - test of disease

Features of squamous dyskaryosis or carcinoma

Features of glandular abnormality

Refer to colposcopy for any abnormality

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19
Q

What might dyskaryosis indicate
(3)

A

CIN
- LSIL (low-grade squamous intraepithelial lesion)
- HSIL (high-grade squamous intraepithelial lesion)

Malignancy

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20
Q

What would features of glandular abnormality indicate

A

CGIN (cervical glandular intraepthelial neoplasm)
Adenocarcinoma

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21
Q

What are the steps in diagnostic cytology

A

Specimen reception
Fixation
Pre-treatments
Preparations (directs, cytospins, LBC)
Staining
Ancillary requests
Reporting

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22
Q

Give an example of a pre-treatment
(2)

A

Need to remove mucous from respiratory sample

May need to centrifuge sample

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23
Q

What kind of prep is done on a LBC

A

Thin prep

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24
Q

What is carried out at specimen reception

A

Minimum data check
All specimens are allocated a unique lab identifier
Gross description (volume/number of slides, colour, consistency)

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25
Q

Why is inspection of samples so important at reception

A

Dictates pre-treatment and preparation technique

26
Q

List some specimen considerations
(8)

A

Collection method
Time lag
Storage
Volume
Consistency
Contaminants
Microbiological hazards (safety cabinet + PPE)
Pre-treatments

27
Q

What contaminants can be found in specimens

A

Blood
Mucous
Clots

28
Q

What are the three main methods of fixation used

A

Alcohol based

Combinations

Formal calcium - lipids

29
Q

What are the two main alcohol based methods of fixation, and when are they used?

A

IMS/ethanol
- cytospin

Methanol
- thinPrep
- air dried

30
Q

What is a combination method of fixation

A

Acetone/methanol

31
Q

What are the benefits of alcohol fixation

A

Rapid alcohol fixation:
- retains nuclear morphology
- retains cytoplasmic structures

32
Q

What is an MGG stain

A

May Grunwald-Giemsa

Can’t do this type of stain on an alcohol fixed sample

Need fresh sample

Need air dried sample so cells will expand

Then fix cells

33
Q

Write about fixation in cytology
(3)

A

Irrespective of preparation method, all samples must be optimally fixed to preserve cells

pre or post-fixation
Relating to staining method
Important to use correct fixative

Only exception is flow cytometry as we need to keep cells viable

34
Q

How do we treat blood contamination
(6)

A

Haemolyse the red blood cells

Use cytolyt - it’s haemolytic

Multiple washes with cytolyte will give you a clean cell pellet

Then transfer your cells to slide

Can use commercial lysing agents

Can remove by use of density gradient -> lymphoprep will give a monolayer of cells (buffy coat)

35
Q

How do we treat mucous contamination

A

Mucolysis

Use Dithiothreitol (DTT) in alcohol

This can be done for MGG staining

36
Q

What are the three ways of preparing samples

A

Cells in suspension -> transfer these to a slide for visualisation

Concentration of cells -> centrifuge to get a cell pellet

Slide preparation methods -> cytospin, ThinPrep, direct spread

37
Q

What does a pink tinge to your sample mean

A

Red cell contamination

Need to do another cytolyte wash

38
Q

How does a cytocentrifuge work?
(4)

A

Cells are alloquated into the funnel

Cells get splattered onto the slide

Filter paper absorbs extra cells

Might do 2 or 3 cytospins for each sample

39
Q

What is the papanicolaou stain and what are the benefits?

A

Alcohol fixation (IMS)

Immediate fixation

Good nuclear morphology

Good cytoplasmic contrast and features

40
Q

What is the May-Grunwals Giemsa stain and what are the benefits?
(6)

A

Air dry prior to fixation
Methanol fixation
Poor nuclear morphology
Good architectural features
Good for small cell populations e.g lymphocytes or small cell carcinoma
Good for amplifying large malignant cells e.g. adenocarcinoma

41
Q

What are the components of the papanicolaou stain and what do they stain?
(3)

A

Haematolylin - nuclear stain

OG6 (Orange 6) -> cytoplasmic stain for keratin/pre-keratin

EA50 (Eosin and light green) -> glycogen and immature cells stained green (protein stain)

42
Q

What are some ancillary techniques?

A

IHC
Cell blocks/clots
Flow cytometry
ISH/FISH
Molecular

43
Q

Give seven IHC stains/groups of stains used?

A

Cytokeratins

BerEP4

S100/HMB45/Melan A

ER/PR Receptor/Her 2 Neu

TTF-1/p63/Chromogranin/PD-L1

Calcitonin/Thyroglobulin

Lymphoid markers

44
Q

What panel of marker is used for melanomas

A

S-100
HMB-45
Melan A

45
Q

What is BerEp4 used for?
(2)

A

Histology stain for diagnosis of basal cell carcinoma

Its an antibody to EpCAM (epithelial cell adhesion molecule)

46
Q

What stains would be used for breast cancer panel

A

ER receptor
PR receptor
Her 2 Neu

47
Q

What is TTF-1

A

Thyroid transcription factor-1

Regulates transcription of genes specific for thyroid, lung and diencephalon

48
Q

What are the advantages of tissue blocks

A

Can take more sections if needed to get a wider IHC panel

Immunocytochemistry can be done -> blocks often used to address QC issues

Long term storage - can go back to sample after years

49
Q

What are the uses of flow cytometry

A

Quantification of lymphocytes
Immunophenotyping
- reactive and lymphoma
- classification of lymphoma

Very useful in Hodgkins and Non-Hodkgins lymphoma as there are many different classifications

50
Q

What is ISH/Molecular
(4)

A

One of the developments in the lab for tumour typing

Uses new advances in genomics

Used for breast, lung (ALK), lymphomas, bladder tumours (UroVysion)

Helps predict response to therapy and risk of recurrence

51
Q

What is ALK

A

Anaplastic lymphoma kinase

52
Q

What are four main aspects of quality control

A

Stain check
Specimen preparation check
Specimen/form checks
Sign out

53
Q

What is IQC

A

Internal control processes and procedure

How the lab ensures quality

54
Q

What is EQC

A

External Quality Control

UKNEQAS e.g. staining and interpretive/reporting schemes

55
Q

How should fresh or preserved specimens be stored

A

In a fridge for up to 1 week

56
Q

How should fixed specimens be stored

A

Can be stored at room temperature but this is dependent on the medium

Retain until reported

57
Q

How should cell blocks be stored

A

Stored long term as tissue blocks

58
Q

How should slides be stored

A

Stored indefinitely

But there will be some deterioration e.g. colour fading

59
Q

What is cell analysis

A

Techniques for preparation and analysis which spans across into non-hospital laboratory setting and research

60
Q

Give some methods of cell analysis
(4)

A

Cell culture work

Morphological analysis of cells

Biochemical analysis

Molecular biology based analysis
- Genomics (DNA/RNA)
- Proteomics

2.2 Cellular Pathology (29 decks)

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