SMEAR & FNAB

Dr Agus Suharto Sp.PA

Bagian Patologi Anatomi FK UMY
Indikasi Smear
THE CONCEPTS Of CELLULAR
PATHOLOGY
1665 : the cell as an entity was first described by Robert Hooke
who recognized the cell structure as a cavity or a vescicle

The term cell, from the Latin cella : hollow space, to

what was thought to be a hollow space.
19th century when the botanist Schleiden in 1838 and
the zoologist Schwann in 1839 revealed that a cell, as a
unit living matter, is formed by the division of another
cell.
The establishment of the cell theory by Schwann was
certainly one of the most remarkable milestones of the
modern scientific discoveries.
 1858 : Rudolpf Virchow, a German pathologist
advanced the concept of cellular pathology.
Virchow’s contribution had thus given morbid
anatomy its most valuable diagnostic tool of
histopathology.

 Disease processes, were investigated in the light of

microscopical observations of cellular changes in
diseases organs.

 Having focused attention on the cell as the unit of

living matter, changes in cell morphology were found
to represent recognizable patterns in pathological
states.
 Some General Principles
 In order to obtain good cytological results a clinician
needs the principles cytology to a good cytologist
and a well-equipped laboratory

 For that reason the efficiency of cytology depends

on 3 factors :
1. The quality of the clinician (his clinical cytology thinking,
his setting indications for cytology, his technique in
obtaining specimens, his handling of the cytologist’s
report, etc)
2. The quality of the cytologist and his personnel and
equipment
3. The quality of the communication between the clinician
and the cytologist
 Degree of Certainty
 It is occur with respect to malignancy

 Initially the description for malignancy were

classified as ‘negative’, ‘doubtful’ and ‘positive’

 For Papanicoloau :
there is a classification to devised cell to
normal, abnormal benign , atypia possibility to
malignant, atypia probability to malignant, and,
malignant cells ( as Class 1,2,3,4,5 respectively)
FALSE POSITIVE – FALSE NEGATIVE

 False positive : when the cytologist

give a definite answer of malignant cell
positive, but it turns to be negative
 False negative : when the cytologist
give a definite answer of malignant cell
negative, but it turns to be positive
 CRITERIA OF MALIGNANCY
The main cytology criteria of malignancy are
found in :
1. The nucleoli
2. The nucleus
3. The cytoplasm
4. The arrangement of cells, either
singly or aggregate
5. The location of cells and the
surrounding material : background
 The Nucleoli
 Abnormality in size
 Irregularity in shape
 The number of nucleoli
 The color of the nucleoli
 The disappears of the nucleoli and the
chromosome
 THE NUCLEUS
 The overlapping / superposition of cytoplasm
 Enlargement : Anisokaryosis usually
accompanied by Anisocytosis, marked
variation in the size of the cells
 Variation in the form
 The number of nuclei per cell
 The chromatin structure of the nucleus
 Mitotic figures
 THE CYTOPLASM
 abnormal amount of cytoplasm
 abnormal form of cytoplasm
 the pattern of cytoplasm
 abnormal colour of cytoplasm
 E.M. cytoplasm alteration
 Phagocytois of the same cell type / Cannibalism
 Cytochemical characteristic of malignancy
THE ARRANGEMENT OF CELLS

 cell aggregates
 multinuclear giant cells
 the rossette
 papillary formation
 mitosis can produce two identical cells :
Twin cells
THE CELLULAR BACKGROUND
 Hemorrhagic

THE NON-CELLULAR BACKGROUND

 Necrosis
 Marbled foamy
 Mucus
 TYPE OF CYTOPATHOLOGY

 EXFOLIATED CYTOLOGY

 NON EXFOLIATED CYTOLOGY - F.N.A.B.

 EXFOLIATED CYTOLOGY
 Specimens consist of single cells or clumps of
cells which are exfoliated,

 and therefore are dissociated from their

surrounding tissues
 The technique is used mainly for the
investigation and diagnosis of malignancy
 The cells are distributed on glass slides, either by
the person who takes the sample, smearing them
directly onto the slide at the time the sample is
taken or, by centrifugation methods in the
laboratory.
 The slides are stained by an appropriate method,
which is the most often the Papanicolaou
technique, and examined by light microscope.
 Since the cells are dissociated from their
surrounding tissue some features that are used in
histopathological diagnosis, such as invasion, and
other architectural abnormalities, are not available
for assessment.
NON EXFOLIATED CYTOLOGY / FNAB
 Cells collected by aspirating through
FINE BORE NEEDLE INTO A SYRINGE
(aspiration cytology)
 Also called aspiration biopsy cytology
 1927 Dudgeon and Patrick proposed the needling of
tumors as a means rapid microscopic diagnosis
 Martin and Ellis, USA at the Memorial Hospital
 1950 Europe particularly Scandinavia : FNA Lopez
Cardozo, in Holland, Zacijek, Sweden then spread to
Japan, Australia, the rest of country
 Procedure
 FIXATION :
After having specimen either from exfoliative or
aspiration methods, the specimen needs to be fixed
by fixation solution in order to get the ideal
morphology of each individual cells before we step to

 STAINING :
to get the optimal morphology of the individual cells
the specimen needs to be stained
 EXFOLIATED CYTOLOGY
Papanicolau procedure :
Cervical smear : After sraping or brushing, the mucin
substance from the cervix can be smeared onto the
surface of slides
Slides : Glass slides ( Object glass ) must be
thoroughly cleaned, dry, and free of grease.
Slides should be labelled before smearing using a
diamond pencil or Slides with frosted ends are
convenient for immediate labelling. After smearing, the
slide should be immersed in fixation sol.at once !
 Fixative
 For routine smears , 96% ethanol
(Absolute alcohol) coviniently in Coplin Jars.
 For Exfoliated cytology from body solution, can be
fixed by : absolute alcohol aa, or aether alcohol aa
For large amount of solution or material,
centrifugation needed in order to get more cells
before smearing
 For FNAB smears : The aspirate can be smeared
between two standard microscope slides. A 0,4mm
haematocytometer coverslip gives better control
over the pressure used in smearing and a more
even spread. Air-dried slides can be transported.
 Other FIXATIVES

 Carnoy’s fixative has the advantage of

lysing red blood cells

 Glutaraldehyde and 10% buffered formalin

should also be available if tissue fragments
for EM or for paraffin embedding are
obtained
 STAINING
 For exfoliative cytology :
Cervical smear : PAPANICOLOAU
Other exf.Cytol.: GIEMSA,
Papanicoloau or H.E. ( block paraffine)
 F.N.A.B. : Giemsa or Papanicoloau or
H.E. (block paraffine)
 EQUIPMENT
Pap’s smear :
gynaecology table/bed
lighting
speculum
ayre spatula or cervical brush
slides with diamond pencil or frosted slide
coplin jar(fixative)
 EQUIPMENT
For F.N.A.B.
Syringes
Syringe Holder
Slides and markeror frosted glass
Fixatives
Sterile containers
Other : skin desinfectans (76%ethanol), sterile
dressings, local anaesthethic (if needed), 76%
alcohol, cotton, plester
In case the mass or the target organ located inside the
body cavity or deep inside, USG or fluoroscopic guiding
is needed.
 EQUIPMENT

MICROSCOPE :
Light microscope
E.M
Fluorescence microscope
 PRAKTIKUM
Smear
Alat dan bahan :
- Tongue Spatle kayu / cotton bud / lidi kapas
- Kaca objek
- Alkohol 96% &/ methanol
- Giemsa Stain
- Mikroskop
+ Cara pengambilan Seperti gambar
+ Fixasi: + 5-15 menit dengan metanol lanjutkan di
cat Giemsa genangi + 5-10 menit
Atau dengan alkohol 96 % + 30 menit
lanjutkan di cat Papaniculou
 FNAB
Alat dan Bahan
Spuit dg Needle : 27 G, 25 G, 23 G
Aspirator (Needle Holder) bila perlu
Methanol
Giemsa Stain
Cara Pengambilan Spesimen
 Cara pengecatan Giemsa
Fixasi : Methanol + 1- 15 menit
Genangi dengan Giemsa 1-5 menit
Keringkan – siap dibaca – tutup dengan
kaca penutup dengan sedikit entelan