Disampaikan dalam acara Workshop Handling & Etika Penggunaan Hewan Coba serta Penandatanganan

MoU Program Studi S1 Farmasi Universitas Gunadarma, Rabu, 20 Februari 2019, Kampus Graha Simatupang,
Universitas Gunadarma

TATA CARA NEKROPSI TIKUS

dan PENGUJIAN PATOLOGI

Prof. Bambang Pontjo Priosoeryanto, Drh, MS, Ph D, APVet, Dipl. ACCM.

Kepala Divisi Patologi
Departemen Klinik, Reproduksi & Patologi (KRP)
Fakultas Kedokteran Hewan; Institut Pertanian Bogor
 Profil Prof. Bambang Pontjo Priosoeryanto, Drh, MS, Ph D, APVet, Dipl. ACCM.

Tempat/Tanggal lahir : Bandung, 28 Februari 1960

NIP : 19600228 198601 1 001
Pangkat : Pembina Utama Madya/ Guru Besar
Golongan : IV/D
Jabatan : Kepala Divisi Patologi Veteriner
Fakultas : Kedokteran Hewan
Departemen : Klinik, Reproduksi & Patologi (KRP)
Divisi : Patologi Veteriner
Alamat Kantor : Jalan Agatis, Kampus IPB Darmaga, BOGOR
Telepon dan Faksimili : (0251) 8421807 – kantor
E-mail : bpontjo4@gmail.com
Strata Tahun Bidang Tempat
S-1 (Drs. Med. Vet.) 1983 Kedokteran Hewan Institut Pertanian Bogor
Dokter Hewan (Drh) 1984 Kedokteran Hewan Institut Pertanian Bogor
S-2 (MS) 1990 Sains Veteriner Institut Pertanian Bogor
S-3 (Ph.D) 1994 Pathology & Preventive Veterinary United Graduate School of Veterinary Sciences,
Medicine Yamaguchi University, Japan
Post Doktoral 1999 Neuropathology of Prion Diseases Institute for Neuropathology, Faculty of Medicine,
Georg-August University, Goettingen, Germany
Brevet 2009 Ahli Patologi Veteriner (APVet.) Asosiasi Patologi Veteriner Indonesia (APVI)
2013 Diplomate Asian College Conservation Asian College Conservation Medicine, Japan
Medicine ( DACCM)
Keahlian Patologi Veteriner
Pemeriksaan Keadaan Luar

Periksa: 1. Keadaan Umum; 2. Jenis kelamin; 3. Kulit dan bulu;

4. Lubang-lubang kumlah; 5. Glandula mamari
Pemeriksaan Glandula Mamaria
Tahap Nekropsi
• Tempatkan hewan dengan
punggung menempel pada
styrofoam
• Fiksir tiap kakinya dengan jarum
pentul.
• Basahi seluruh kulit dan bulu
bagian abdomen dan medial
kaki dengan alkohol
• Buat sayatan di kulit sepanjang
linea alba, mulai dari ujung
dagu (regio mentalis) hingga ke
tepi anterior tulang pelvis
(pecten ossis pubis).
Penyayatan Kulit
Pembukaan kulit
• Kulit dipreparir hingga dapat
dikuakkan ke samping tubuh,
termasuk kulit di bagian atas
dari kaki-kaki.
• Fiksir kulit dengan jarum
pentul
• Sambil membuka kulit,
dilakukan pengamatan pada
subcutan
Lokasi Sistem Limfatik
Tikus Betina Dengan Glandula Clitoris (GC: Gl. Clitoridis)
Pembukaan Ruang Abdomen
• Otot perut (dinding abdomen)
digunting di linea alba,
dimulai dari ujung tulang
dada (processus xiphoideus)
hingga pecten ossis pubis.
• Gunting otot perut dibawah
kurvatura tulang rusuk dan di
daerah sekitar paha, hingga
otot-otot perut dapat
dikuakkan ke kanan dan ke
kiri
• Untuk lebih memudahkan
mengamati organ-organ
rongga perut, otot-otot perut
disingkirkan.
Pemeriksaan Ruang Abdomen

Pembukaan ruang
abdomen dan
pemeriksaan umum
Rongga Perut dengan Berbagai Organ
Sistem Pencernaan dan Limpa
Pemotongan oesophagus (OE) agar dapat mengeluarkan
organ-organ pencernaan. CX: Cartilago xiphoidea; Spleen
Sistem Pencernaan dan Limpa
Lambung Tikus dan Bagian-bagiannya
Ginjal dan Kelenjar Adrenal
 Urogenitalia Tikus Jantan

Penis dikeluarkan bersama dengan kelenjar asesorius (gl. vecicularis, gl.

vesicula seminalis, gl. prostate, gl. bulbourethralis), vesica urinaria, urethra
dan penis. DD: Ductus deferens.
Testis, Epididimis dan Ductus deferens
Urogenitalia Tikus Jantan
Urogenitalia Tikus Jantan

Kelenjar asesorius pada tikus jantan. P: gl. Prostate, GC: gl.

coagulationis, GV: gl. Vecicularis, GB: Gg. Bulbourethralis,
U: Urinary bladder.
Alat Reproduksi Tikus Betina

Organ-organ di ruang abdomen dan pelvis tikus betina. R:

Rectum, OV: Ovarium, SP: Symphysis pelvina.
Pembukaan Rongga Dada
1.Tulang rusuk terakhir
dipotong ke depan
menuju arkus tulang
sternum
2.Pemotongan dilakukan
pada sisi kanan maupun
kiri, kemudian perlekatan
dengan difragma
dipreparir, sehingga
tulang sternum dan
sebagian tulang rusuk
berbentuk segitiga dapat
dibuang
Pembukaan Rongga Dada

CX: Cartilago xiphoidea. DI: Diaphragma.

Pemeriksaan Timus
Pengeluaran Organ-organ Rongga Dada

Pengeluaran organ-organ rongga dada dalam satu

rangkaian: lidah, esophagus dan trakhea
Pembukaan Kranium
• Buat sayatan pada
kulit kepala tepat
dibagian tengah dan
berakhir sejajar
telinga
• Buang kulit dan otot-
otot di bagian dorsal
dan kaudal kranium
Pembukaan Kranium
dan Pengambilan Otak
Pembukaan Kranium
 Pemeriksaan
gl. Harderian

Pemeriksaan
hyphophysa
setelah otak
diambil
 Pemeriksaan Kelenjar Ludah

Pemeriksaan kelenjar ludah dan limfonodus. LNN: Lymph nodes.

GSL: gl. sublingualis. GSM: Gl. submaxillaris.
 Pengambilan Mata

GLE: Gl. lacrimalis extraorbitalis. GP: Gl. parotis

 Bagian Ventral Leher

GT: gl. Thyreoidea, L: Larynx, T: Trachea, M: Musculus masseter.

Nervus ischiadicus (NI) diantara otot paha medial
Sampling Organ
 How To and How Not To Submit your Biopsy Specimens
DA Kamstock, DVM, DACVP, LL Debuse, EJ Ehrhart, DVM, DACVP, BE Powers, DVM, DACVP
Colorado State University Veterinary Diagnostic Laboratory

1 Clinical Information 3 Packaging 6 Denoting Margins

 Formalin filled jars containing specimens should be placed in
a plastic bag, box, or other container with absorbent material to
absorb any leakage (A)
Please provide signalment and pertinent clinical information  Paperwork should be placed in a separate plastic bag to avoid
 Certain lesions occur more commonly in different species and contact with formalin if leaking does occur. Such contact can
certain breeds result in altered and illegible paperwork (B)
 Anatomical location of a lesion, as well as clinical appearance and  Your fresh sample size should never be larger than the most
progression, may also be critical information to allow your narrow portion of the jar in which you are submitting it (C). If it
pathologist to provide you with the best possible diagnosis and/or is, this will require cutting plastic jars or breaking glass jars
differentials (undesirable) in order to retrieve the tissue which may become
altered in the process.
 If you have a specific question, are concerned about a possible
A Tumor Bed Samples
disease process, or have a list of differentials you’d like to rule out,
please indicate such.
 Again, please make every effort to provide this necessary
information in the designated areas on the CSU-VDL biopsy B
NO
submission form. It will help us help you help your patients.
YES
Surgical Ink 1. Ink the area of interest
2. Be sure to ink prior to bread loafing (if
needed) 3. Allow ink to
begin drying before placing the specimen in
formalin
Tagging
1. Used to indicate margins or for orientation
2. Use variable numbers and/or colors of suture
3. Provide a clear description on the
submission form denoting what the sutures
indicate (i.e. one suture = cranial margin)
1. Submission of samples from the post-surgical
bed 2. Any tumor in these
specimens is evidence of remaining microscopic
C disease 3. Similar to
“submitting multiple sites” clearly label and submit
each region individually

4 Submitting Multiple Sites 7 Other Things to Avoid

YES NO The optimal method by which to submit NO
multiple lesions from a single animal is to Please help keep our technicians
submit each specimen individually in its own fingers safe and DO NOT submit
2 Tissue Fixation respective and appropriately labeled jar. This specimens with needles for any
reason!
should be reflected on the submitted
paperwork.
 Routine tissue fixation = 1:10 tissue to neutral buffered formalin YES Please do no staple, suture, or pin
tissue to cardboard. It can damage NO
 For appropriate fixation, 0.5 – 1.0 cm tissue thickness is If multiple specimens are submitted in a tissue and prevent appropriate margin
recommended single container (which is less ideal) there assessment
suture
 Bread loafing (incomplete parallel cuts at a minimum of 2cm needs to be some method of tissue
identification (i.e. suture) to denote
What Else Should I know?
apart) can be performed on large specimens (be sure to avoid
complete transection or too many cuts which can both result in specimens relative to respective anatomic 8
loss of tissue orientation!) YES site.
It is important for you to realize that after all is said and done
 Specimens can be held to fix (at least 24 hours) at your clinic the pathologist typically evaluates 1 to 4, 5um thick sections
prior to sending to the lab to avoid shipping large volumes of
formalin which can be expensive and increase the risk of leaking
5 Endoscopic Biopsies from the entire specimen which is submitted (A).

A The optimal method by which to
A. Incomplete parallel cuts at a
minimum of 2cm apart (bread
loafing) can be utilized to assist
with appropriate tissue fixation
for large specimens
YES Our staff and pathologists are here to assist you. If you have
NO NO
questions about how best to submit your sample or have
questions regarding any other issues, please contact the lab -
(970)-297-1281.
Additional information on the CSU-VDL can be found on the
web at www.dlab.colostate.edu Visit Us!
B C Do not submit endoscopic
Do not place endoscopic submit an endoscopic biopsy is to
biopsies wrapped in gauze A. From Our staff
biopsies on fragments of place it in a screen cassette after
B. Large samples can be held C. This is an example of an ~20cm sponges. Specimens may
cardboard. Specimens will which the cassette should be placed in mass to is here
and fixed at your clinic prior to diameter mass lesion which was fixed become lost or may be crushed an appropriately labeled formalin filled working
during the attempted retrieval
either float off or, if adhered, block to
submitting to the lab to help at the clinic and subsequently sent to tissue will slough off during jar. If individual cassettes are labeled hard for
avoid shipping large volumes of the lab in a plastic, labeled, zip lock process. It is better to submit properly (sharpie or no2 pencil),
slide
retrieval and/or may be you!
formalin which may be costly and bag devoid of any formalin. (bar = the specimen free floating in the multiple cassettes can be place in one
associated with cardboard
hazardous ~2.5 cm) jar than with gauze or any other jar.
fibers
material
 Sampling Organ
• Mengumpulkan sampel se-aseptik mungkin
• jaringan
• aspirat
• kultur darah

– Alat nekropsi steril

• Disinfeksi cold pack
• Alat autoclave
• Alkohol and pembakar

• Masukkan sampel yang representatif

• Ambil sampel dari yang rusak
• Ambil sample dari yang segar dan baik
Sampling
Organ

• Fiksatif Buffered Neutral Formalin 10%

• Volume Fiksatif : Organ = 10 : 1
• Waktu fiksatif yang cukup minimal 48
jam sebelum prosesing jaringan
 JARINGAN
Jaringan
• Koleksi sebesar genggaman (jika kurang, jaga kelembaban)

• Hanya jaringan yang dikirim untuk histopatologi yang harus diberi

formalin

• Jaringan dapat dikirim dalam:

– Whirl Pac
– Mangkok fecal/ urine yang bersih (belum digunakan) atau
kontainer plastik.
– Snap cap tube
– Red top tube

• Jaringan harus tetap lembab

– (cukup untuk melindungi jaringan)
– Bungkus dalam steril 4X4 soaked dalam salin steril
Contoh Pengiriman
Sample yang Buruk
Contoh Pengiriman Sample yang Baik
 PENGUJIAN PATOLOGI
MAKROSKOPIS MIKROSKOPIS
Patologi Anatomi Histopatologi
Sitopatologi
Histokimia (Pewarnaan Khusus)
Imunohistokimia (Reaksi Ag-Ab)
Elektron Mikroskopi (SEM & TEM)
ANALISI YANG DIPAKAI
1. Deskriptif
2. Skoring Lesio dengan metode Histoskor (Intensitas x
Distribusi), yang dilanjutkan dengan Analisis Statistika
 Studi Infeksi Virus
Parvovirus - Myokarditis
Intranuclear inclusion body

Sarang radang
 Analisis
Histopatologi
Tahapan
Kerusakan
otot jantung
akibat
Kardiotoksin
 Studi Histopatologi
Kinetika Tumor

TC

Basal Sel Tumor

HP
 Studi
Transplantasi
Sel Tumor

Pada Nude
Mouse
Studi Tumor Imunologi
 Imunohistokimia
Antibodi-Antikeratin
 Penulisan Laporan

• Signalemen
• Anamnese
• Temuan Patologi Anatomi
• Histopatologi
• Diagnosa/ Diferensial diagnosa
REFERENSI
• Henrik Elvang Jensen
Department of Veterinary
Pathobiology,
Royal Veterinary and Agricultural
University, Copenhagen, Denmark
• Vincenzo Covelli - Guide to the
Necropsy of the Mouse
• Dan lain lain
Terima Kasih