Types of sample:

1. Conventional Pap smear (CP)

2. Liquid-Based preparation (LBP)
i. Thin preparation
ii. Sure path

Sample adequacy:

CP- 8K to 12K squamous cells (well-preserve, well-visualized)

Indicate sufficient cell, at
LBP- 5K to 20K squamous cells ( )
the correct site including
At least 10 glandular cells (endocervical or squamous metaplastic cells) transformation zone (TZ)

*metaplastic will not be counted as glandular neoplasm

Organisms to look for: bacteria, virus,
Gynae Negative for intraepithelial malignancy (NILM) parasite, fungal

Epithelial cell abnormalities (ECA) non-neoplasm

Cell reactive: inflamed, colour change,
cytoplasm change, nucleus change

Tumor/neoplasm/new growth benign: new growth that will not metastasize from its
origin in-situ)

malignant: Cell metastasize from its origin through blood

vessel or lymphatic
Usually, virus : LSIL HPV, but can also be

LSIL w/out HPV infection

NILM change not due to infection/non-neoplasm is caused by

Intrauterine Contraceptive Device (IUCD)

Radiation cause changes in DNA

Atrophic smear
Due to decreased hormone level (give local
- After menopause oestrogen treatment and asked to repeat in
- Lactation three months)

* Atrophic smears are difficult to analyse and a repeat Pap smear will ensure the most accurate result

Menopause more immature cells because no hormone release

Squamous mature cell superficial

P/E high
Intermediate
Squamous immature cell parabasal

Basal

Recognize and understand the characteristics of cells

Superficial, intermediate, parabasal derived from basal cell

Superficial squamous cells i. Outermost layer of non-keratinizing

epithelium
ii. Centrally located pyknotic nucleus
iii. No nuclear detail can be seen due to
nuclear degeneration.
iv. Polygonal shaped
v. Eosinophilic coloured cytoplasm
vi. Flat
vii. Abundance during the late
proliferative and ovulatory phases of
the menstrual cycle (estrogen at
peak)
Intermediate squamous cells i. Centrally located nucleus
ii. The nucleus is vesicular with fine
evenly dispersed granular chromatin
iii. Cytoplasm is thin, transparent, and
typically stains basophilic
iv. Abundance during the luteal and
early follicular phases of the
menstrual cycle, and the second and
third trimester of pregnancy
(progesterone at high)

Parabasal squamous cells i. Found in basal layer

ii. The nuclear detail reveals a finely
granular chromatin
iii. High N:C ratio
iv. Round to oval shaped
v. Dense homogenous basophilic
cytoplasm
vi. Uncommon finding, often seen in
patients who lack estrogen
(premenstrual, post partum, taking
estrogen-restricting hormones or
postmenopausal)
Squamous metaplastic cells i. Arise from the basal layer of
glandular epithelium (as a protective
response to stimuli, creating the
transformation zone)
ii. Round centrally located nuclei
iii. Round to polygonal shaped
iv. Dense biphasic staining cytoplasm

Endocervical glandular cells i. Round basally placed nucleus

ii. Fine granular chromatin with an
occasional nucleoli.
iii. Tall, columnar-shaped
iv. The mucinous cytoplasm are
visualized as being granular or
vacuolated
v. Can be seen on Pap tests in three
arrangements:
- single cells,
- as strips,
- as a sheet
*arrange in uniform manner (single @ palisading)
*honeycomb pattern
*normally cilia can be seen
Endometrial glandular cells i. Round- to oval- to bean-shaped
nucleus
ii. The nuclei are hyperchromatic
iii. Small and cuboidal shaped
iv. In cycling women, endometrial cells
are expected to be seen on Pap
tests from the first day bleeding
starts through the twelfth day.
v. After day twelfth day, the presence
of endometrial cells may be
considered a significant finding
*densely arranged have to look at peripheral side
*randomly arrange not honeycomb pattern
*3-D, ball-like shape
*seen within @ 2-3 days after menstruation

Chromatin in nucleus indicate actively dividing cells, which has high risk of leading to
carcinoma, seen in metaplastic
endocervical
carcinoma
Method to screen: horizontal vertical
The grid part of the slide should be on the left side

Lab 17/3/17

WBCs (purple color)

Bacteria

= shift in flora suggestive of bacterial vaginosis

Normal flora of vagina Doderlein bacilli

No visible thin line squamous cell is being

eaten by the bacteria
Parabasal

Glycogen in the cells

Glycogen in cell, can be seen like rough

surface in the cytoplasm (only in intermediate
& parabasal only!)
Condition:
1. post-partum
2. lactation

Endocervical cells

Honeycomb pattern
arranged in uniform manner:
1. single
2. palisading (basally located nucleus)

Cilia is normally seen, from terminal bar (thin

line)
Lab 31/3/17

Trichomonas vaginalis
(can see the nucleus inside cell)

Perinuclear halos

4 characteristics of fungal infection:

Abundant WBCs
Canon-ball
Shish kebab
Perinuclear halos

Candida sp.
(the hyphae)

= NILM with fungal infection

Negative for Intraepithelial Lesion or Malignancy (NILM)

1) Fungal consistent with Candida spp.

Candida grows rapidly; needs glycogen for energy. Thus, it grows on the cells.

The budding yeast may be misdiagnosed with RBC since both stain eosinophilic (red)

How to differentiate rbc with budding yeast?

yeast cell appear in big cluster

fine focusing to see rbcs central parlour

Repeat smear after antibiotic treatment

2) Bacteria vaginosis

Gardenella vaginalis (coccobacilli)

Commensal bacteria = Doderlein bacilli

Only 1 of these organism can present on the smear

GV DB
X = shift of normal flora
X

GV grows on the squamous cell @ covered by a layer of GV = clue cells

Repeat smear after antibiotic treatment

3) Actinomyces spp.

Eosinophilic to purplish colour

Filamentous projection @ wooly-body

Associate with IUCD usage

Repeat smear after antibiotic treatment

4) Virus

Effect:

o Multinucleation presence of multiple nucleus in a cell

o Margination chromatin move to peripheral side @ central clearing = ground glassappearance
o Moulding

Cell that always affected by Herpes Simplex Virus (HSV) = endocervical

Repeat smear in 3 to 6 months

4) Trichomonas vaginalis

Characteristics of TV:

o Nucleus towards the end

o Pale colour (ghost cell)
o Cytoplasm membrane foamy
o Cyanophilic coloured cytoplasm
o Basophilic coloured nucleus

Bacteria associate = Leptotrix

Repeat smear after antibiotic treatment

Other Non-Neoplastic Findings

5) Inflammation

4 characteristics:

o Abundant WBCs
o Canon-ball Inflammation
o Perinuclear halos
o Slightly high N:C ratio / enlarge nucleus

+ signage to search for any entity. If absent, = inflammation

Repeat smear in 3 to 6 months

If no entity; Repeat smear as scheduled

6) Atrophy normal for > 40 y/o women

Characteristics:

o Parabasal flat monolayer sheet (immature cell due to lack of hormone)

o Uniform size of nucleus / enlarge nucleus
o Smooth nuclear membrane
o Fine and evenly distributed chromatin

*ovary secrete the estrogen & progesterone

Causes:

o Menopausal state
o Total hysterectomy

Atrophy + Inflammation = tick both

Also presence of smudge cell

If still present after treatment, should consider HSIL

Repeat smear after hormonal treatment

7) Irradiation

Person diagnosed with squamous cancer

To monitor treatment successful or not

If succeed, may see regression of carcinoma cell

Characteristics:

o Bizarre cell shape

o Cellular debri
o Degenerated blood
o Cytomegaly
o Nucleomegaly
o Engulfed PMN

8) IUCD

Small cluster of cell with vacuolation = endometrial

Being degenerated

Very high N:C ratio

Lab 7/4/17

Background: watery diathesis

High N:C ratio

Trichomonas vaginalis

Trichomonas vaginalis (on cell)

o Nucleus eccentric
o Pale colour
o Cytoplasm cyanophilic,
membrane foamy

Background:filmy

Clue cells

Gardnerella vaginalis

Actinomyces - Clumps of filamenteous

organism
Herpes Simplex Virus
o Multinucleation
o Margination
o Moulding

Ground glass appearance

Glandular endocervical cell in single

Candida albicans

Pseudohyphae

Shish kebab

Perinuclear halos

Budding yeast
Turolopsis glabrata

Candida without pseudohyphae

High N:C ratio

Other sign of inflammation:

Biphasic (2 colours in a cell)
Lab 14/4/17

Trichomonas vaginalis with presence of blood

Blood (eosinophilic colour)

Bacteria vaginosis

Watery diathesis
-a layer in cyanophilic color
-looks like flow of water
-protein substance smeared in the background

Tumor diathesis
-old blood (eosinophilic) swarming
-pyknotic dots = fragmented nucleus
Lab 20/4/17

Koilocytes
e protein from hpv
Wht type of e protein

Single koilocytes
Hyperchromatic
compare colour with wbc

Pleomorphic nucleus
Thick and irregular nuclear membrane
Koilocyte
Binucleation (inflammation changes)
Pearl formation (inflammation changes)
Non keratinizing squamous carcinoma
LSIL
Parakeratosis
sq cell with lots of keratin
very mature
Orangeophilic colour
Wht is hyperkeratosis

Coarse chromatin
Thick nucleus

Classical koilocytes
2-3 times enlargement
Coarse chromatin
Atypical cell
Hyperchromatic
Very high NC ratio
SQUAMOUS METAPLASTIC HSIL
Cytoplasm spreading, larger than
endocervical
Centrally located nucleus

HSIL
Overlapping
Cytoplasm quite big
Nuclear groove

Binucleation
LSIL
Classical HSIL
Scanty cytoplasm
Hyperchromatic
Coarse chromatin
No nuclear groove

Nuclear groove

NILM
parabasal
Fine granular chromatin
Nuclear enlargement
 Karyohexis (nucleus remnant)

Tumor diathesis consist of

old rbc
Fragmented nucleus
Wbc
Proteinaceous

elongation of nucleus
Wbc inside cytoplasm (polymorphs
ingestion)

Pleomorphism of nucleus
 Orangeophilic cytoplasm
Fibrious/ projectile
NKSCC

HSIL
Suggestion: Refer to gynaecologist oncologist

Lab 27/4/17

flat
Abundant cytoplasm
Spiky cytoplasm, cannot be
defined/knobby
No nuclear membrane thickness
Slightly hyperchromatic
Coarse n clumping chromatin
ASCH
Management: Colposcopy n biopsy
30% of agcnos is hsil

high nc ratio
Presence of blood
Irregular shape
Coarse Chromatin clumping
No nucleoli
Thick nuclear membrane
HSIL without tumor diathesis

Homogenise look?
 Polymorph ingested cytoplasm
Basally located nucleus (glandular
abnormality)
Nucleoli
Losing polarity, dispersed
Distinct cytoplasm
Background: proteinecious residual
AGCNOS

Glandular;
palisading
Vacuolation inclusion

Macronucleoli
 the nucleus about to form feathering
characteristics
Hyperchromatic
Macronucleoli
Nucleus round to oval
AGCNOS Favour neoplastic due to lack of
AIS characteristics

elongated nucleus

flat
Sharing community border

Refer to: benign

smooth community border