INTRODUCTION TO CYTOLOGY

AND GYNAE CYTOLOGY

DR WAN ROHANI WAN TAIB
DBB40103
History of Cytology

Dr. George Papanicolaou

introduced the concept that
diagnosis of malignancy could be
achieved by microscopic in
1928 .Then, in 1943, he and the
gynecologist Herbert F. Traut
published a report entitled
"Diagnosis of uterine cancer by
the vaginal smear."
Cytology
To identify abnormal cells
To identify some inflammatory conditions and
causative agents
To identify some non-neoplastic changes such as
repair, metaplasia and effects of hormones

Before you can recognise cancerous or other abnormal cells

must have good knowledge of normal cells at the relevant site
3 Types of Cytology
 Gynaecological
Pap smear, cells exfoliated and collected by brushing or scraping
 Non-Gynaecological (General)
cells exfoliated from organs, collected by scraping, washing or
brushing epithelial surfaces (eg bronchial brushings or washings)
cells exfoliated into fluid and extracted from fluid
(eg urine, sputum, CSF)
 Fine Needle Aspiration
cells collected by using fine needle
(eg breast lump, lymph node, liver, lung)
Gynae Cytology FNAC

Non-Gynae Cytology
Non Gynae Cytology
Bronchial brushing/washing
Pleural Fluid
Peritoneal Fluid.
Pericardial Fluid
Urine
Sputum
Cerebrospinal Fluid
Others body fluids.
Staining:
Papanicolaou stain (PAP) – nuclear detail
May Grunwald Giemsa stain (MGG) – cytoplasmic detail
Fine needle aspiration for Cytology (Fnac)
Swelling or mass
Breast
Thyroid
Lymph node
Neck
Others.
Gynae Cytology
Cervical smear – Pap smear
Vaginal smear – Vault smear
Staining – Papanicolaou
Reporting system – Bethesda 2014 (KKM)
Getting a Pap test
What is pap smear?
The Pap smear is a simple medical procedure used
to detect early changes in the cells of the cervix
which might become cancerous.  
The procedure was named after Dr George
Papanicolaou, the physician who developed the
procedure, after he discovered in 1928, that
changes in the appearance of cells in the cervix can
serve as a warning sign that these cells may
develop into cancer.  
What is pap smear?
The procedure involves collecting a sample of cells
from the cervix and the vagina, and sending the cell
sample to the laboratory to be examined for abnormal
cells that may grow into cancer.
Pap smear only screening test for the prevention of
cervical cancer
 Patient instructions
Intercourse is not recommended the night before
appointment.
A woman should have a Pap smear when she is not
menstruating.
The best time for screening is between 10 and 20 days
after the first day of her menstrual period.
For about two days before testing, a woman should avoid
douching or using spermicidal foams, creams, or jellies or
vaginal medicines (except as directed by a physician).
These agents may wash away or hide any abnormal
cervical cells.
Making A Pap Smear

Sample smeared on clean

glass slide
Immediate fixation in 95%
alcohol
Anatomy of the Female Genital Tract
Vagina
Uterus
Fallopian tubes
Ovaries
The graphic above depicts the
major components of the female
reproductive tract.
1. Vagina - female sex organ,
passageway for menstrual
blood and fetus during
childbirth

2. Cervix - region connecting the

uterus to the vagina; the
muscles of the cervix support
the weight of the fetus during
pregnancy
Anatomy FGT
3.Uterus (womb) - location where
fertilized egg develops into a
fetus and is nourished until birth.
Note that the walls of the uterus
are thick and lined with muscles
4.Fallopian (uterine) Tube -
location of fertilization of an egg
by a sperm cell
5. Ovary - produce and store
gametes (eggs) and produce the
female sex hormones, estrogen
and progesterone
Normal Cervix
Squamous Epithelium
Superficial

Intermediate

Parabasal
Basal
Squamo-Columnar Junction
Junction of pink cervical skin
and red endocervical canal
Key portion of the cervix to
sample
 Area of metaplastic change
in the cervix
 Most cancers arise in the TZ.
Whom to screen?
 Screening starts at age of 20-25 –continues over
symptomatic postmenopausal women
 Screening women with family history of cervical
carcinoma or patients diagnosed with in situ lesions
 Screening and follow-up on known cases of
carcinoma/ radiotherapy/post-hysterectomy status –
Vault smear
 Pap smear screening is recommended for those who
have been vaccinated against HPV
 To evaluate hormonal assessment of vaginal smear.
 Sampling cervico-vaginal smears

Evaluation requires optimal sampling, smearing,

fixation, staining
Midcycle, less blood or inflammatory cells (not during
menstruation)
Combined cervico-vaginal smear widely used
Different Types of Brushes : spatula,
broom, cytobrush
 Techniques of sampling

Cervico-vaginal sampling
Endocervical sampling
Vaginal aspiration
(irrigation)

Method of smearing
1.Conventional smear method
2.Liquid based method
Samples are collected from the outer
opening or os of the cervix using
 An Ayre’s spatula – conventional
PAP
 Endocervical brush or a plastic-
fronded broom – Liquid Based
Cytology
Conventional Smear Method
Liquid Based Cytology
(LCB) Method
Liquid Based Cytology (LCB)
 Since the mid-1990s, techniques based around placing the sample into a vial
containing a liquid medium that preserves the cells – media: primarily ethanol-based

 Works on different principle to ultimately give accurate results

 Sensitivity and specificity has high range of 90%

Types of LCB:
 Thin-prep: filtration and collection of vacuum-packed cells on membrane and
transferring to slide
 Sure Path: centrifugation and sedimentation through a pressure gradient

Uses of LBC
 Gynae cytology
 Non-gynae cytology eg thyroid cyst fluid, pleural effusions, urine, brushing samples
etc
The ThinPrep Pap test was designed to address many of the sampling pitfalls associated
with the conventional Pap test. Studies prove that the ThinPrep system is superior in
effectiveness at detecting abnormalities.

Unlike the conventionally prepared Pap test slide, studies show the ThinPrep slide is
clean and truly representative of a woman’s state of cervical health, making it a more
accurate test. Laboratory professionals are able to interpret the results with greater
confidence.
The entire procedure takes about 70 seconds per slide and results in a
thin deposit of cells in a circle 20 mm in diameter
 Advantages
Conventional Papanicolaou test
 The gold standard for 50 years
 Comfort
 Ready for staining when it arrived in the
lab
Liquid-Based Cytology
 Virtually all cellular material is collected
 Ease of use for smear taker
 Limited obscuring material
 Even distribution of cells
 Fewer but well-preserved cells to
examine
 Ease of examination, decreased
screening time required
 Microorganisms preserved
 Extra slides can be made, including cell
blocks and immunocytochemical/DNA
probe
 Additional molecular testing performed
on residual vial material
Disadvantages
 >80% of the cellular material is
discarded after sampling
 300,000 cells screened (needle in
haystack)
 Uneven distribution of cellular material,
clumping/overlapping of cells, fixation
artifacts, unsatisfactory smears.
 New technology causes insecurity
 Training of clinicians, lab technicians,
cytotechnologists and pathologists is
required
 Increased workload in lab preparations,
monotony in preparing slides
 Fixatives (shipping, disposal)
 Increased costs (lab preparation labor
and disposable)
 Requires some vigilance, intensity of
screening.
SCREENING AND STAINING?????