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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
Why is inspection of samples so important at reception
Dictates pre-treatment and preparation technique
26
List some specimen considerations (8)
Collection method Time lag Storage Volume Consistency Contaminants Microbiological hazards (safety cabinet + PPE) Pre-treatments
27
What contaminants can be found in specimens
Blood Mucous Clots
28
What are the three main methods of fixation used
Alcohol based Combinations Formal calcium - lipids
29
What are the two main alcohol based methods of fixation, and when are they used?
IMS/ethanol - cytospin Methanol - thinPrep - air dried
30
What is a combination method of fixation
Acetone/methanol
31
What are the benefits of alcohol fixation
Rapid alcohol fixation: - retains nuclear morphology - retains cytoplasmic structures
32
What is an MGG stain
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
Write about fixation in cytology (3)
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
How do we treat blood contamination (6)
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
How do we treat mucous contamination
Mucolysis Use Dithiothreitol (DTT) in alcohol This can be done for MGG staining
36
What are the three ways of preparing samples
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
What does a pink tinge to your sample mean
Red cell contamination Need to do another cytolyte wash
38
How does a cytocentrifuge work? (4)
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
What is the papanicolaou stain and what are the benefits?
Alcohol fixation (IMS) Immediate fixation Good nuclear morphology Good cytoplasmic contrast and features
40
What is the May-Grunwals Giemsa stain and what are the benefits? (6)
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
What are the components of the papanicolaou stain and what do they stain? (3)
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
What are some ancillary techniques?
IHC Cell blocks/clots Flow cytometry ISH/FISH Molecular
43
Give seven IHC stains/groups of stains used?
Cytokeratins BerEP4 S100/HMB45/Melan A ER/PR Receptor/Her 2 Neu TTF-1/p63/Chromogranin/PD-L1 Calcitonin/Thyroglobulin Lymphoid markers
44
What panel of marker is used for melanomas
S-100 HMB-45 Melan A
45
What is BerEp4 used for? (2)
Histology stain for diagnosis of basal cell carcinoma Its an antibody to EpCAM (epithelial cell adhesion molecule)
46
What stains would be used for breast cancer panel
ER receptor PR receptor Her 2 Neu
47
What is TTF-1
Thyroid transcription factor-1 Regulates transcription of genes specific for thyroid, lung and diencephalon
48
What are the advantages of tissue blocks
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
What are the uses of flow cytometry
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
What is ISH/Molecular (4)
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
What is ALK
Anaplastic lymphoma kinase
52
What are four main aspects of quality control
Stain check Specimen preparation check Specimen/form checks Sign out
53
What is IQC
Internal control processes and procedure How the lab ensures quality
54
What is EQC
External Quality Control UKNEQAS e.g. staining and interpretive/reporting schemes
55
How should fresh or preserved specimens be stored
In a fridge for up to 1 week
56
How should fixed specimens be stored
Can be stored at room temperature but this is dependent on the medium Retain until reported
57
How should cell blocks be stored
Stored long term as tissue blocks
58
How should slides be stored
Stored indefinitely But there will be some deterioration e.g. colour fading
59
What is cell analysis
Techniques for preparation and analysis which spans across into non-hospital laboratory setting and research
60
Give some methods of cell analysis (4)
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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