Specialty Guide · Path-iQ Global Pathology Review

Cytopathology
The Complete Guide 2026

Cytopathology is the examination of individual cells and small cell clusters for diagnostic purposes. Its greatest strength is speed and minimally invasive sampling — a fine needle can aspirate a deep-seated lesion under ultrasound guidance in minutes, yielding a diagnosis that would otherwise require major surgery. Cytopathology underpins cervical cancer screening programmes that have reduced cervical cancer mortality by over 70% in countries with high uptake.

Updated 30 July 2026 · Path-iQ Editorial · About Path-iQ →

Sample Types in Cytopathology

Sample TypeCollection MethodCommon ApplicationsProcessing
Fine needle aspiration (FNA)22–25G needle, US-guided or palpation-guidedThyroid, lymph node, breast, salivary gland, pancreas, liver, lungDirect smears + cell block or LBC
Cervical cytology (Pap smear)Cervical brush → LBC vial or direct smearCervical cancer screening; HPV triageLBC (ThinPrep/SurePath) or conventional
Urine cytologyVoided midstream urineUrothelial carcinoma detection/surveillanceLBC + Paris System reporting
Sputum cytology3 early-morning deep-cough specimensSuspected central lung carcinoma (squamous, SCLC)Saccomanno fixative; Papanicolaou stain
Bronchial wash/BALBronchoscope flush of airwaysLung carcinoma, Pneumocystis jirovecii, MACCentrifugation; Diff-Quik + Pap stain
EBUS-FNAEndobronchial ultrasound-guided FNAMediastinal staging; sarcoidosis; lymphomaRapid on-site evaluation (ROSE) + cell block
EUS-FNAEndoscopic ultrasound-guided FNAPancreatic lesions, submucosal GI tumoursROSE + cell block; molecular ancillary
Pleural/peritoneal fluidThoracocentesis or paracentesisMalignant effusion detection; mesotheliomaCytocentrifuge; cell block for IHC
CSF cytologyLumbar punctureLeptomeningeal carcinomatosis; CNS lymphoma; medulloblastomaCytocentrifuge within 1 hour of collection

Reporting Systems — Bethesda and Beyond

The Bethesda System for Cervical Cytology (TBS)

The Bethesda System (last revised 2014) standardised cervical cytology reporting into six categories that link directly to clinical management algorithms. The system uses a single terminology for both conventional smears and liquid-based cytology (LBC).

TBS CategoryDefinitionApproximate CIN2+ RiskRecommended Management
Negative for intraepithelial lesion or malignancy (NILM)No abnormal squamous or glandular cells<1%Routine screening interval
ASC-USAtypical squamous cells of undetermined significance5–10%HPV co-test; colposcopy if HPV positive
ASC-HAtypical squamous cells — cannot exclude HSIL25–40%Colposcopy
LSILLow-grade squamous intraepithelial lesion (HPV/CIN1)15–25%Colposcopy
HSILHigh-grade squamous intraepithelial lesion (CIN2/3)70–80%Colposcopy + biopsy; LEEP/cone if CIN2+
AGC / AISAtypical glandular cells; adenocarcinoma in situVariable; AIS 48–72% CIN2+ riskColposcopy + ECC + endometrial sampling
Squamous cell carcinomaMalignant squamous cellsUrgent colposcopy + biopsy; oncology referral
AdenocarcinomaMalignant glandular cells (endocervical, endometrial, extrauterine)Urgent colposcopy + biopsy; oncology referral

Bethesda System for Thyroid Cytology (TBSRTC)

The Bethesda System for Reporting Thyroid Cytology (3rd edition, 2023) classifies thyroid FNA into six categories with associated malignancy risks and management recommendations.

CategoryBethesda CategoryMalignancy Risk (ROM)Usual Management
INon-diagnostic / unsatisfactory5–10%Repeat FNA with ultrasound guidance
IIBenign0–3%Clinical follow-up; repeat ultrasound
IIIAtypia of undetermined significance (AUS/FLUS)10–30%Repeat FNA; molecular testing (ThyroSeq, Afirma)
IVFollicular neoplasm / Hürthle cell neoplasm25–40%Lobectomy or molecular testing
VSuspicious for malignancy60–75%Lobectomy or total thyroidectomy
VIMalignant97–99%Total thyroidectomy

Other Standardised Reporting Systems

SystemOrgan/SampleEditionPublished By
The Paris System (TPS)Urine cytology2nd ed. 2022International urine cytology working group
Papanicolaou Society of Cytopathology (PSC) PancreaticobiliaryPancreas, bile duct, ampulla2nd ed. 2020PSC working group
Milan System (MSRSGC)Salivary gland FNA2nd ed. 2022International salivary gland cytology group
International System for Serous Fluid Cytopathology (ISfSFC)Pleural, peritoneal, pericardial fluid1st ed. 2020IAC-ISFC working group
Yokohama SystemPancreatic cyst fluidRevised 2024IAP working group
RCPath/IAC Urinary CytologyUrine, bladder washUpdated 2023RCPath / IAC joint committee

Liquid-Based Cytology vs Conventional Smears

Liquid-based cytology (LBC) transfers the cervical sample into a preservative vial (ThinPrep/Hologic or SurePath/BD). A thin layer of cells is prepared from the vial for microscopic examination. LBC offers several advantages: fewer unsatisfactory samples (1–2% vs 5–8% for conventional smears), the ability to test the residual sample for HPV and other molecular assays, better cell preservation, and a cleaner preparation. Most European and Australasian national screening programmes have transitioned entirely to LBC; the United States uses both LBC and conventional smears.

Rapid On-Site Evaluation (ROSE)

ROSE is the practice of having a cytotechnologist or cytopathologist immediately evaluate a preliminary stained smear during an FNA procedure to confirm specimen adequacy and preliminary diagnosis. ROSE has been shown to reduce non-diagnostic rates in EBUS-FNA and EUS-FNA of pancreatic lesions from 15–20% to under 5%, and improves overall diagnostic yield. It is standard of care for EBUS-guided mediastinal staging in many high-volume centres.

Leading Cytopathologists — Global 2026

Syed Z. Ali
Thyroid & EBUS Cytology
Johns Hopkins University School of Medicine

One of the architects of the Bethesda System for Reporting Thyroid Cytology. Editor of multiple editions of the TBSRTC atlas and an authority on thyroid FNA adequacy criteria.

Philippe Vielh
Breast & Gynaecological Cytology
Institut Gustave Roussy, Paris

Former Secretary General of the International Academy of Cytology (IAC). Expert in breast FNA, liquid-based cytology standardisation, and molecular ancillary testing in cytopathology.

Ritu Nayar
Cervical & Thyroid Cytology
Northwestern University Feinberg School of Medicine

Lead editor of The Bethesda System for Reporting Cervical Cytology (4th edition). Expert in HPV-related cervical disease and the evolution of cervical cancer screening algorithms.

Martha Pitman
Pancreaticobiliary Cytology
Massachusetts General Hospital

Lead author of the Papanicolaou Society of Cytopathology System for Reporting Pancreaticobiliary Cytology. Expert in EUS-guided FNA of pancreatic lesions and cyst fluid cytology.

Ashish Chandra
Urinary & Pulmonary Cytology
Guy's & St Thomas' NHS Foundation Trust, London

Expert in urinary tract cytology, pulmonary FNA, and the Paris System for urine cytology. Editor of the RCPath cytology curriculum and a leading figure in UK cytopathology education.

Esther Diana Rossi
Thyroid & Head & Neck Cytology
Fondazione Policlinico Gemelli, Rome

Authority on thyroid cytology, salivary gland FNA, and the application of molecular testing to Bethesda III and IV thyroid nodules. A contributor to the Milan System for Salivary Gland Cytology.

Frequently Asked Questions

What is the difference between cytology and histology?
Histology (histopathology) examines intact tissue architecture — the organisation of cells within stroma and their relationship to each other. Cytology examines individual cells or small clusters stripped of their architectural context. Cytology is faster, less invasive, and can be performed on samples (urine, sputum, effusions) that cannot yield a tissue biopsy, but it provides less information about invasion, margins, and tumour architecture than a histological section.
How accurate is thyroid FNA?
In experienced hands, thyroid FNA has a sensitivity of 85–95% and specificity of 90–98% for malignancy. The Bethesda III (AUS/FLUS) and IV (follicular neoplasm) categories represent the diagnostic challenge — approximately 20–30% of Bethesda III nodules and 25–40% of Bethesda IV nodules prove malignant on surgical resection. Molecular testing panels (ThyroSeq v3, Afirma Genomic Sequencing Classifier) refine risk stratification for these indeterminate categories.
Is cervical screening changing with HPV primary testing?
Yes. Many high-income countries (UK, Australia, Netherlands) now use hrHPV DNA testing as the primary screening test, with cytology triage only for HPV-positive results. This approach is more sensitive than cytology-first screening and allows longer screening intervals (5 years for HPV-negative individuals). The United States uses HPV co-testing (HPV + Pap smear simultaneously) or HPV-primary testing, with guidelines from ASCCP 2019 based on risk stratification.

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