THE CYTOLOGICAL FEATURES

OF NORMAL EPITHELIAL
CELLS.
ANATOMICAL PATHOLOGY 2B
Course Code: ANP621S
Compiled by: S. Israel
Revised by: BR Tsauses
August 2022
 Approach to this presentation
• Use these slides as a summary and revision of theory and practical
covered in class.
• The information provided in these slides is a great combination of
theory and practical.
 INTRODUCTION
• Identification of normal epithelial cells is the first step in learning how to screen
cervical samples.
• The Papanicolaou stain is ideally suited for this purpose, containing a
combination of cytoplasmic stains and a nuclear stain, allowing the
identification of the various types of epithelial cells.
• The cytoplasmic stains are: eosin, which stains superficial cells pink; light green,
which stains the cytoplasm of the less mature cells; and orange G, which stains
keratinized cells orange.
• The nuclear stain is haematoxylin, which stains the nuclei blue-black.
• The full range of epithelial cells can be identified by their morphology and
staining properties.
 CONTINUE…
• Before describing the specific features of each cell type, you should be aware of
the descriptive vocabulary that cytologists use.
• At the most basic level, the distinction between normal and neoplastic epithelial
cells is based primarily on the morphology of nucleus.
• Correlations between cytology and histology have identified the following
features as the most useful for distinguishing normal and neoplastic cells:
• Absolute nuclear size, usually expressed as a diameter measurement.
• Relative nuclear size, i,e. the ratio of the size of the nucleus to the size of the cell.
• This is referred to as the nucleocytoplasmic ratio, or NCR for short.
•
CONTINUE…
Chromatin pattern- the combination of proteins and deoxyribonucleic
acid(DNA) within the nucleus.
• Chromatin patterns refers to the degree of the chromatin granules as
seen under the light microscope.
• The term pyknotic is used to describe a particularly dense chromatin
pattern, whereas vesicular is the term used to describe a fine granular
chromatin pattern.
• Chromatin may also be coarsely granular or there may be discernible
pattern, in which case the nucleus is described as ‘bland’.
 CONTINUE…
• Chromatin distribution
• Describes the variation in the spatial density of chromatin granules
over the area of the nucleus.
• Although image analysis systems can measure this precisely,
cytologists find this attribute of the most difficult to interpret and
define.
• Put crudely, it is usually sufficient to describe chromatin distribution as
either uniform or irregular.
• Nuclear shape e.g. round, oval, irregular.
• Regularity of the nuclear membrane, e.g. smooth, wrinkled, angular
• Intensity of nuclear staining, e.g. normochromasia,
hyperchromasia(darkly stained), and hypochromasia(pale staining)
 PARABASAL CELLS

• PARABASAL CELLS
• Round/oval cells 12-30 um in diameter.
• Dense (heaped) cytoplasm.
• Rounded/oval nucleus~8 um in diameter, occupying half of the cell.
• Evenly distributed vesicular chromatin.
• Associated with atrophy
• Naked nuclei- delicate cytoplasm
• Clusters of naked parabasal nuclei mimic endometrial cells
 INTERMEDIATE CELLS

• INTERMEDIATE CELLS
• Polygonal cells 30-40 um in diameter, sometimes with folded cells edges.
• Cytoplasm usually cyanophilic, sometimes eosinophilic.
• Round/oval nucleus~ 8um in diameter.

• Low NCR .
• Fine vesicular chromatin
 Predominance associated with progesterone, 2nd half of cycle and pregnancy
• Key reference
 Size
 Stain, texture
 SUPERFICIAL CELLS

• SUPERFICIAL CELLS
• Polygonal cells 35-45 um in diameter.
• Eosinophilic cytoplasm.
• Low NCR
• Small pyknotic nucleus, 5 um in diameter.
 Largest squamous cell in Pap smear
 Voluminous thin cytoplasm
 Pyknotic (ink dot) nucleus
 Predominance associated with estrogen, reaches peak at mid cycle
 The Glandular Epithelial Cells
• Cells from the endocervix – endocervical cells
o Pallisade arrangement
o Honeycomb arrangement
• Cells from the endometrium – endometrial cells
o 3D balls – “top hat” arrangement
o Stromal endometrial cells
 ENDOCERVICAL CELLS

• ENDOCERVICAL CELLS
• Honeycomb sheets, palisaded strips, or loosely associated single columnar cells.
• Finely vacuolated cyanophilic cytoplasm, sometimes filled with clear mucus.
• Note the uniformity, orderly = honeycomb appearance
• Nuclei round/oval with smooth outlines – benign glandular epithelium
• On right- seen from the side- palisades – note uniform, basally located nuclei
• No stratification
 CONTINUED…

• Ciliated endocervical cells, terminal bars and cilia

• Pencil thin, elongated endocervical cells – Lugol’s iodine application at time of colposcopy may
cause this change
• Cilia occasionally visible at one end.
• Oval nucleus with fine chromatin pattern, occasionally with one or more small nucleoli.
• Nuclei sometimes have nipple-like protrusions.
 METAPLASTIC CELLS
• METAPLASTIC CELLS
• Morphology depends on degree of maturation.
• Cells size approximately that of parabasal/early intermediate cells.
• Usually cyanophilic cytoplasm.
• Cytoplasmic projections, giving rise to the term spider cells.
• Nuclei are vesicular, variable in size, with small nucleoli.
 CONTINUED…
• Metaplasia is the replacement of one mature cell type
with another mature cell type
• Delicate endocervical glandular cells replaced by more
robust squamous cells
• Process begins with reserve cell hyperplasia and
metaplastic cells often seen with endocervical cells
• These cells originate from the squamocolumnar junction
o Immature and mature squamous metaplasia
METAPLASTIC CELLS…
• Small round or
polygonal metaplastic
cells with slightly
enlarged N/C ratio.
• Smooth nuclear
membranes.
• Chromatin pattern is
finely granular and
evenly distributed.
• Small round nucleoli
are commonly seen.
 METAPLASIA BEGINS WITH…
• Reserve cells similar to basal cells – begin to proliferate – RCH
• Capacity to differentiate into glandular or squamous cells
• RCH at point begins to differentiate – first immature then
mature squamous metaplastic cells
• The reserve cell thought to be source of squamous, adeno +
adenosquamous Carcinoma.
IMMATURE SQUAMOUS
METAPLASIA
• Parabasal size cells,
cobblestone
arrangement, thick
dense cytoplasm
looks like its been
cut out of
cardboard.
• -endo/ectoplasmic
rim common feature
of immature
metaplasia
SPIDER CELLS

These are cells

from immature
squamous
metaplasia,
forcibly scraped
from the
mucosal surface.
METAPLASIA
• Mature squamous metaplasia
– resemble normal
intermediate cells, except
their outlines may be a little
more rounded than polygonal
and cytoplasm may be slightly
denser than mature
intermediate cells.
• Eventually squamous
metaplasia matures
completely and becomes
cytologically indistinguishable
from the native squamous
epithelium.
Deep stromal
invaginations
of endocervical
mucosa – 8mm
deep, termed
endocervical
glands, not true
glands
Relatively
mature
squamous
metaplastic
epithelium
bridging an
invagination of
the
endocervical
mucosa.
Complete
blocking leads
to mucus filled
cysts – Naboth
follicles
RESERVE CELL HYPERPLASIA
• Small basaloid cells, high
N/C ratio, delicate
undifferentiated cytoplasm
– rarely seen on cytology
• Clue may be cells are
associated with
endocervical cells or
immature metaplasia
METAPLASIA
• Normal polygonal
squamous metaplastic
cells
• Nuclei – round to oval,
bland chromatin.
• Identify that TZ has been
sampled – 10 well
preserved endocervical or
metaplastic cells required.
 ENDOMETRIAL CELLS
• ENDOMETRIAL CELLS
• Normally present up to 12 day of cycle
• Round/oval cell clusters with dense core of stromal cells and periphery
of larger epithelial cells.
• This type cell clustering is sometimes referred to as wreath or top hat
formation.
• Sometimes present as tight groups of disorganized cells with crumpled
nuclei.
• Individual cells 8-10 um diameter with little cytoplasm.
• Hyperchromatic nuclei same size as intermediate cell nucleus.
ENDOMETRIAL CELLS…

Endometrial cells
tend to be packed
together and
degenerate –
normal endometrial
cell is the size of an
intermediate cell
nucleus.
CONTINUED…

Classic
“double
contour” – or
“Top Hat”
arrangement
-stroma in
center
surrounded
by epithelium
– associated
with the
exodus.
 STROMAL ENDOMETRIAL CELLS
Deep stromal - round to spindle
shaped with small oval nuclei
and scant cytoplasm

Superficial stromal
endometrial cells – form
loose aggregates
NAVICULAR CELLS
•Glycogenated cell
•Associated with pregnancy,
postpartum, lactation
•Samples often contain a large
number of glycogenated
intermediate cells (navicular cells)
which can mimic viral infection.
 REFERENCE
• Shambayati B, 2011,Cytopathology.