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ROSE SSA Lung—EBUS/EUS mediastinal

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Images first published: Glinski et al. Single slide assessment: A highly effective cytological rapid on?site evaluation technique for endobronchial and endoscopic ultrasound?guided fine needle aspiration.  Cytopathology 30, 164–172 (2019).

There are many personalised therapies available for advanced lung carcinomas. ROSE SAA is vital in this area to establish that the correct diagnostic cell yield is obtained.

Tumour typeApproved tests required for targeted therapyYield
Squamous cell carcinomaImmunocytochemistry panel
PDL1 – Programmed Death Ligand 1
PDL1 – minimum of 100 viable cells (Be careful of necrotic lesions). Caution: Ensure that atypical single cells are not present within the necrotic debris
AdenocarcinomaImmunocytochemistry panel
PDL1 – Programmed Death Ligand 1
EGFR – Epidermal Growth Factor Receptor
ALK – Anaplastic Lymphoma Kinase
ROS 1 – c-ros oncogene 1
Approximately 50% Tumour load to ensure there is enough yield for EGFR (Epidermal Growth Factor Receptor) and ALK (Tyrosine Kinase receptor) testing. In accordance with referral centre protocol
PDL1 – minimum of 100 viable cells (Be careful of necrotic lesions)
Small cell carcinomaImmunocytochemistryLocal ICC requirements – dependant on panels used
CarcinoidImmunocytochemistryLocal ICC requirements – dependant on panels used
LymphomaLarger immunocytochemistry lymphoma panels
Flow cytometry
If possible, use 19 / 20 Gauge procore needle to ensure there are large cores for IHC panels – 
Metastatic carcinomaImmunocytochemistry and any molecular testing required i.e. Melanoma – BRAFLocal ICC requirements – dependant on panels used
Lymph node samplingNot applicableMinimum of 30% lymphocyte population (More than peripheral blood)
Presence of Lymphoglandular bodies
Lympho-histocytic aggregates
Adequacy criteria guidanceRequirement
Morphological appearance EGFR – Approximately 50% Tumour load  to ensure there is enough yield for EGFR (Epidermal Growth Factor Receptor) and ALK (Tyrosine Kinase receptor) testing. In accordance with referral centre protocol
Lung AdenocarcinomaPDL1 – minimum of 100 viable cells (be careful of necrotic lesions)
Necrotic samplesCaution: ensure that atypical single cells are not present within the necrotic debrisAt ROSE necrotic slides can be misinterpreted as cellular. Review on high power to ensure the cells are viable.
Lymphoma casesIf possible, use 19 Gauge procore needle to ensure there are large cores for IHC panels. An abundance of lymphocytes is not seen in benign node sampling is seen at ROSE. In some cases, highly atypical cells are also present.

Images first published: Glinski et al. Single slide assessment: A highly effective cytological rapid on?site evaluation technique for endobronchial and endoscopic ultrasound?guided fine needle aspiration.  Cytopathology 30, 164–172 (2019).

Diagnostic pitfalls and considerations

  • Contaminant bronchial sampling
  • Necrosis, particularly in SCC. Ensure that only viable cells are assessed.
  • Process all of the residual material in the laboratory even if the ROSE SSA is inadequate. Subsequent diagnostic material can be found in the induced clot of the residual material. Particularly evident in metastatic renal cell carcinoma where there can be an excess of blood due to the nature of the tumour.
  • Streamlined approaches to immunocytochemistry panels must be adopted in laboratory practice for non-small lung carcinoma, to ensure material is not unnecessarily wasted. Standard ICC panel – TTF1, p40 and CK5/6.
  • When multiple sites are sampled it crucial that the specifics of the lesion are captured on the form and the corresponding vial and slides – For example in EBUS – Node station numbers are used.  Involvement of certain stations can lead to different treatment pathways and prognosis. If staging is being conducted – check that the least suspicious node is sampled first to prevent false positives or inform clinician to use a new needle. Renew all consumables used for ROSE SSA to prevent contamination between samples.
  • Fibrotic change due to recent radiotherapy can make it difficult to obtain a cellular sample. Iatrogenic changes must also be taken into consideration.