Winda Nur Pebriani Yusup, Amd.

AK
 Sitologi  ilmu yang mempelajari tentang sel
 Diagnostik Sitologi  interpretasi dari sel

Non-
Expoliated
Expoliated

Prinsip teknik sitologi

Proses spesimen sitologi  preparat refresentatif 

bisa diinterpretasikan & didiagnosa.
The first era
- Abad19
- Sel-sel kanker yang terlepas (exfoliated)
dapat ditemukan di semua jenis spesimen

-1861
-Pada sekresi faring
-Post mortem
-Keratinizing squamous cell carcinoma
The second era

-wet fixation
 Pulasan Papanicolaou
- screening pada kanker serviks

Titik awal Perkembangan

Sitologi

Dr. George N Papanicolaou (1832-1962)

The third era
- technique of FNAC
- Diagnostic cytology and its
histopathologic basis

The fourth era

- the Bethesda system of
Dr. Leopold G Koss
reporting cervical/vaginal
cytology diagnoses
 Kelebihan Pemeriksaan Sitologi

 Mudah
 tidak
 Murah menimbulkan
stimulasi
 Cepat metastase
 Sederhana
 Efektif untuk
 Resiko minimal diagnosis tumor
 Waktu pemeriksaan saluran cerna, paru,
saluran kemih, dan
singkat lambung
Kekurangan Pemeriksaan Sitologi
 Diagnosis  berdasar perubahan sitoplasma dan

inti sel

 Perubahan  bukan akibat kesalahan teknis

 Mendeteksi lesi  di permukaan mukosa

Kekurangan Pemeriksaan Sitologi

 Lesi  tidak tertutup keratin tebal

 Tidak efektif pada lesi hiperkeratotik  sel-sel

abnormal tertutup oleh lapisan keratin

 Masih perlu konfirmasi biopsi

 Seringkali bahan yang terambil tidak

refresentatif
 LCS
Vaginal smear/ cervical smear
Sputum
Sendi
Bronchial washing/ brushing
Urine cerebrospinal

Cairan lambung
pleura
Aspirasi jaringan tumor

Imprint jaringan tumor ascites

Cairan tubuh lain pericardium

Exfoliative
cytology

Fine Needle
Aspiration
Cytology (FNAC)
  Sel yang terlepas atau diambil dari permukaan Epitel
dari berbagai organ.

wash smear

scraping brushing
Scraping
Pleura Pericardium
CSF Ascites
 Fiksasi 
- mencegah proses degenerasi sel dan jaringan

- mempertahankan morfologi sel

 Wet Fixation/ fiksasi basah 95% Ethyl Alcohol
(Ethanol)

 Air drying/ Fiksasi kering

 Giemsa/ Diff Quick

 Liquid-based Fixation for

Papanicolaou Tests
• Spray Fixation

Jarak antara spesimen - spray fixation.

 Sediaan apus kering sebelum difiksasi

 Fiksasi tidak menggunakan alkohol 96%

 Hairspray/ spray fixation  jarak terlalu dekat

Fiksasi secepatnya PENTING

 I. Proses sediaan Cairan Tubuh/ Body Fluid

 Identifikasi Sampel dan formulir

 Sentrifuge  sedimen apusan  fiksasi alkohol
96%

SENTRIFUGE
Teknik Apusan  Pull Apart
 Cytocentrifugation/ Sedimen + NaCl 0.09% (1:1)
Cytospin

Wet
Fixation 1000 rpm 5 menit
Papanicolaou

Dry
Fixation
Giemsa
Cytocentrifugation/
Cytospin

• to concentrate cells within a defined area

• a filter card between the chamber sample and the glass
slide resulting in cell to slide adhesion

Hydraulic force

Centrifugal force
 Shandon cytocentrifuge I

 Shandon cytocentrifuge II

 Shandon cytocentrifuge III

 Shandon cytocentrifuge IV

 Wescor cytopro

 Hettick cytocentrifuge

 Leif’s centrifugal cytology buckets

 II. Proses sediaan Sputum

Teknik Apusan SPREADING

- used for thick mucoid secretions

- smears of fresh sputum and bronchial aspirates
- Fiksasi alkohol 96 %
III. Proses sediaan Aspirasi/ FNAB

Dry Air Alkohol 96 %

Diff Quick
Fixation (wet Fixation)

Giemsa Methanol Papanicolaou

Diff Quick Giemsa Papanicolaou

Diff Quick Papanicolaou Giemsa

 IV. Proses sediaan Pap Smear / Cervical
Smear
konvensional

Pap smear
Alkohol 96 % Papanicolaou
konvensional

collection of
Saline moistened
samples
Ayre spatula cotton tip
applicator

Conventional smear
Liquid based cytology

 Cytologi Liquid medium

 Cells are collected from cervix(any other
site)  placed directly into liquid
preservative,  transferred to slide.
 Sample is processed and resultant thin
smear  easy to screen
 Cellprep LBC system liquid based
cytology system that used blowing
technology.
 Filtration and collection of vacum packed
cells on a membrane and transferring to the
coating slide.
 Cell prep kit

Specimen filtration Membrane Microscope

& Preservation Vial Cervix Brush
Filter slide
Cell prep
Conventional smear Cellprep slide
Conventional smear Cellprep slide
 Convetional ThinPrep Cellprep

Fixation Ethanol methanol Ethanol

Collection Smear on slide Sample rinsed Collection

in vial device left in
vial

Cell sample Random Uniform Uniform

distribution distribution distribution
over 20 mm of over 20mm of
slide slide
Collection EC brush, EC brush, Cervex-Brush
device spatula, spatula, most effective,
Cervex- Cervex-Brush tip
Brush rinsed deposited into
in vial vial

Preservation Air drying, blood, All preservation All preservation

artifacts inflammation, artifacts greatly artifacts greatly
irregular reduced reduced
distribution of
cells
Automate Not Vacuum Blowing
d applicable pressure technology
processing through
TransCyte filter

Imaging Not applicable Available Available

technology

Ancillary Not applicable HPV, Chlamydia, HPV, Chlamydia,

testing gonorrhea gonorrhea
Papanicolaou

Giemsa

Diff Quick
Papanicolaou Staining

 Ditemukan  Dr. George N. Papanicolaou

(1832-1962)

 Pewarnaan polikromatik

 Menunjukkan berbagai variasi morfologi sel 

sesuai derajat maturitas dan aktifitas

metaboliknya
 Prinsip Papanicolaou

Hidrasi dan Dehidrasi

Hidrasi

Dehidrasi

Dehidrasi dan clearing solution

 Zat Warna
Papanicolaou

Nuclear staining/ Harris

pewarnaan inti Hematoxylin
 Two cytoplasmic counter staining

1. Orange
G -6

2. Eosin
including three
Azure (EA)- stains
50 –Eosin Y
–Light Green
–Bismarck
brown
 Prinsip Pewarnaan Papanicolaou

(1) Fixation
- 95% ethyl alcohol
- minimum of 15 minutes

(2) Nuclear staining

Harris haematoxylin
(3) Cytoplasmic staining
(4) Dehydration
- Rinse the smears in absolute alcohol for
two or three changes for the removal of
water.
(5) Clearing
- alcohol is being replaced with Xylene
(6) Mounting
 fixation

hydration

dehydration

clearing
 Age of dyes
Type of fixatives No. Of slides in
each dye

Moisture and h
umidity
Regressive or
progressive Factors affecting
Pap staining

Length of
staining time

Presence or absence of in
Quality of cell sa flammatory cell changes
mple
 Giemsa

Tujuan  Detail morfologi inti sel

1. Larutan Giemsa
Bahan 2. Larutan phosfat buffer
(ph 6,8)
3. Methanol
Prosedur Pewarnaan Giemsa

1. Fiksasi methanol  5 menit

2. Keringkan di udara
3. Tetesi dengan pewarna Giemsa dengan
perbandingan (GZ: Buffer phosfat 1:4)
4. Cuci dengan aquadest  kering di udara
5. Mounting
 Diff Quick

 Fast three-step differential

stain for bone marrows,

smears, FNAs, and

Includes:
microorganisms such as H.
Reagent 1: Fixative;
pylori
Reagent 2: Eosin;

Reagent 3: Methylene Blue

 Prosedur Diff Quick

1. Apus sampel pada slide

keringkan di udara

2. Celup Reagen #1 5 kali

3. Celup Reagen #2 5 kali

4. Celup Reagen #3 5 kali

5. Bilas dengan aquadest

6. Keringkan di udara/ dryer

RESULT
1. Naylor B, Ramzy I. Cytopathology:the past, the present and the glimpse
into the new millenium. In. Gray W, Mckee GT. Diagnostic cytopathology.
2nd edition. Churchill Livingston.2002. p. 3-13.

2. Bales CE. Laboratory techniques. In. Koss LG. Koss’s diagnostic cytology
and its histopathologic bases. 5th edition. Lippincott Williams and Wilkins.
2005. p. 1570- 1634.

3. Weidmann JE, Keebler CM, Fasik MS. Cytopreparatory techniques. In.

Bibbo M, Wilbur DC. Comprehensive cytopatholgy. 3rd edition. Saunders.
2008. p. 835-58.
4. Cibas ES. Cervical and vaginal cytology. In. Ducatman BS. Cytology:
diagnostic principles and clinical correlates.4th edition. Saunders. 2014. 1-57

5 . Orell SR, Veilh p. The techniques of FNA cytology. In. Orell SR, Sterret GF.
Fine needle aspiration cytology. 5th edition. Churchill Livingstone. 2011. p.
8-27.