Pre analitik spesimen

histopatologi
Didik Setyo Heriyanto

Department of Anatomic Pathology, Faculty of Medicine, Public Health,

and Nursing – Dr. Sardjito Hospital, Universitas Gadjah Mada
 Laboratory processes encompass three phases:
the preanalytical, analytical and postanalytical

Preanalytical1 Analytical1 Postanalytical1

Sample Collection Sample Processing and

Sample Transport Sample Storage Testing Reporting Record Keeping
and Stabilization Management

Graphic adapted from World Health Organization. apps.who.int/iris/bitstream/10665/44665/1/9789241548274_eng.pdf.

1. World Health Organization. apps.who.int/iris/bitstream/10665/44665/1/9789241548274_eng.pdf. Accessed June 2020
 The greatest scope for laboratory errors exists during the
pre-analytical phase of the workflow

Preanalytical1 Analytical1 Postanalytical1

Sample Collection Sample Processing and

Sample Transport Sample Storage Testing Reporting Record Keeping
and Stabilization Management

60-75% of all laboratory errors may occur in the pre-analytical phase,

specifically those related to specimen collection, handling,
transportation, preparation, and storage2,3

Graphic adapted from World Health Organization. apps.who.int/iris/bitstream/10665/44665/1/9789241548274_eng.pdf.

• 1. World Health Organization. apps.who.int/iris/bitstream/10665/44665/1/9789241548274_eng.pdf. Accessed January 31, 2019. 2. Lippi G, et al. Clin Chem Lab Med. 2015;53(3):357-370. 3.
Compton C. Int Clin Pathol J. 2016;2(4):94-95.
 Nurse

Clinician
The most common
problems that occur in the
pre-analytic phase are
FIXATION AND FIXATIVE

Laboratory
assistant
 Teknisi senior banget

Konsulen PA Konsulen PA
Senior Senior

Konsulen PA Muda

PPDS Senior

PPDS Junior
Sebelum lanjut ke proses fiksasi, beberapa aspek berikut ini harus
diperhatikan:

1. Nama pasien
2. Usia pasien
3. Asal regio diambilnya spesimen
4. Tujuan pemeriksaan
5. Cek apakah fiksasi sudah sesuai dan volume telah sesuai
6. Jika spesimen sangat besar dan spesimen membutuhkan cairan
fiksasi yang banyak, segeralah kontal ahli patologi untuk membuat
potongan awal
FIXATION is a process for preserving of tissue in solution to retain the “life-like” state. It can
then undergo histological review in Anatomical Pathology (AP).

Most people think fixation is a simple process of storing tissue in liquid after surgery, but it is actually
a series of complex
Principle of fixation
Fixative has a property of forming CROSS LINKS
between proteins, hence keep them in their exact
locations

The crosslinking of most of the proteins will

continue to occur yielding complete fixation of
the tissue.

FIXATIVE
Cold ischemia time
Time between tissue removal from patient body
and fixation of specimen

Prolonged cold ischemia time results in

preferential loss of signals that may lead to false
negative results

American Society of Clinical Oncology-College of

American Pathologists (ASCO – CAP)
recommends one hour or less
 Preservative agent
FIXATIVE
Neutral buffer formalin 10% is most commonly used for tissue fixation

Confusing nomenclature: formaldehyde, formalin, or

10% NBF?
§ Formaldehyde is GAS.
§ 100% formalin is saturated (37%) formaldehyde solution.
§ 10% NBF is buffered formalin that contains part of concentrated formalin and water thus containing
approximately 3.7% formaldehyde.

Dilution in water Dilution in water

+ Salt buffer

Formaldehyde (Gas) Concentrated formalin solution 10% Neutral Buffered Formalin

(37-40% Formaldehyde)
FIXATIVE Why Use Buffered Formalin?

• The physiological pH of cell is 7-7.5

• Unbuffered formalin solution will oxidize to

form formic acid (low pH) à react with Hb

à acid formaldehyde hematin à artifact of

brown-black pigment deposited

https://www.leicabiosystems.com/knowledge-pathway/fixation-and-fixatives-2-factors-influencing-
chemical-fixation-formaldehyde-and-glutaraldehyde/
FIXATIVE
How to make 1L 10% formalin from 40%
formalin?

C1.V1 = C2.V2

40.V1 = 10.1000

V1 = 250 Q3 TRUE OR FALSE?

Diambil 250ml (dari 40% stok
formalin) dicampur dengan
750ml air menjadi formalin 10%
1liter
3. Apakah volume cairan formalin 40% sebanyak 250ml
sesuai untuk membuat larutan (pengenceran) formalin 10%?

C1.V1 = C2.V2

40.V1 = 10.1000 A. BENAR

V1 = 250
B. SALAH

Diambil 250ml (dari 40% stok

formalin) dicampur dengan
750ml air menjadi formalin 10%
1liter
FIXATIVE How to make NBF 10% 1L?

+ +

100ml Formalin
(37-40% stock solution) 4g NaH2PO4 6.5g Na2HPO4 900 ml
water
FIXATIVE
How to make 1L 10% formalin from 40%
formalin?

C1.V1 = C2.V2

40.V1 = 10.1000

V1 = 250 TRUE OR FALSE?

Diambil 250ml (dari 40% stok
formalin) dicampur dengan
750ml air menjadi formalin 10%
1liter
FIXATIVE
How to make 1L 10% formalin from 40%
formalin?

C1.V1 = C2.V2 C1.V1 = C2.V2

40.V1 = 10.1000 100.V1 = 10.1000

V1 = 250 V1 = 100

Diambil 250ml (dari 40% stok Diambil 100ml (dari 40% stok
formalin) dicampur dengan formalin) dicampur dengan
750ml air menjadi formalin 10% 900ml air menjadi formalin 10%
1liter 1liter
FIXATIVE Ratio specimen : fixative
FIXATIVE Ratio specimen : fixative

Ratio specimen : fixative

1: 10-20
Specimen size: 1X1X0.5 = 0.5 cm3 (= 0.5 ml)

1 X 1 X 0.5 cm
Volume of fixative required 5-10 ml