tata cara nekropsi tikus dan pengujian...
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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
Disampaikan dalam acara Workshop Handling & Etika Penggunaan Hewan Coba serta PenandatangananMoU Program Studi S1 Farmasi Universitas Gunadarma, Rabu, 20 Februari 2019, Kampus Graha Simatupang,Universitas Gunadarma
Profil Prof. Bambang Pontjo Priosoeryanto, Drh, MS, Ph D, APVet, Dipl. ACCM.
Tempat/Tanggal lahir : Bandung, 28 Februari 1960NIP : 19600228 198601 1 001Pangkat : Pembina Utama Madya/ Guru BesarGolongan : IV/DJabatan : Kepala Divisi Patologi VeterinerFakultas : Kedokteran HewanDepartemen : Klinik, Reproduksi & Patologi (KRP)Divisi : Patologi VeterinerAlamat Kantor : Jalan Agatis, Kampus IPB Darmaga, BOGORTelepon dan Faksimili : (0251) 8421807 – kantor
E-mail : bpontjo4@gmail.com
Strata Tahun Bidang TempatS-1 (Drs. Med. Vet.) 1983 Kedokteran Hewan Institut Pertanian BogorDokter Hewan (Drh) 1984 Kedokteran Hewan Institut Pertanian BogorS-2 (MS) 1990 Sains Veteriner Institut Pertanian BogorS-3 (Ph.D) 1994 Pathology & Preventive Veterinary
MedicineUnited Graduate School of Veterinary Sciences, 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 Medicine ( DACCM)
Asian College Conservation Medicine, Japan
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
• 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).
Tahap Nekropsi
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
Penyayatan Kulit
Lokasi Sistem Limfatik
Tikus Betina Dengan Glandula Clitoris (GC: Gl. Clitoridis)
• 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.
Pembukaan Ruang Abdomen
Pembukaan ruang abdomen dan pemeriksaan umum
Pemeriksaan Ruang Abdomen
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
Penis dikeluarkan bersama dengan kelenjar asesorius (gl. vecicularis, gl. vesicula seminalis, gl. prostate, gl. bulbourethralis), vesica urinaria, urethra
dan penis. DD: Ductus deferens.
Urogenitalia Tikus Jantan
Testis, Epididimis dan Ductus deferens
Urogenitalia Tikus Jantan
Kelenjar asesorius pada tikus jantan. P: gl. Prostate, GC: gl. coagulationis, GV: gl. Vecicularis, GB: Gg. Bulbourethralis,
U: Urinary bladder.
Urogenitalia Tikus Jantan
Organ-organ di ruang abdomen dan pelvis tikus betina. R: Rectum, OV: Ovarium, SP: Symphysis pelvina.
Alat Reproduksi Tikus Betina
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.
Pembukaan Rongga Dada
Pemeriksaan Timus
Pengeluaran organ-organ rongga dada dalam satu rangkaian: lidah, esophagus dan trakhea
Pengeluaran Organ-organ Rongga Dada
•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
Pembukaan Kraniumdan Pengambilan Otak
Pembukaan Kranium
Pemeriksaan gl. Harderian
Pemeriksaan hyphophysa setelah otak
diambil
Pemeriksaan kelenjar ludah dan limfonodus. LNN: Lymph nodes. GSL: gl. sublingualis. GSM: Gl. submaxillaris.
Pemeriksaan Kelenjar Ludah
GLE: Gl. lacrimalis extraorbitalis. GP: Gl. parotis
Pengambilan Mata
GT: gl. Thyreoidea, L: Larynx, T: Trachea, M: Musculus masseter.
Bagian Ventral Leher
Nervus ischiadicus (NI) diantara otot paha medial
Sampling Organ
4 Submitting Multiple Sites
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
3 Packaging
Routine tissue fixation = 1:10 tissue to neutral buffered formalin
For appropriate fixation, 0.5 – 1.0 cm tissue thickness is
recommended
Bread loafing (incomplete parallel cuts at a minimum of 2cm
apart) can be performed on large specimens (be sure to avoid
complete transection or too many cuts which can both result in
loss of tissue orientation!)
Specimens can be held to fix (at least 24 hours) at your clinic
prior to sending to the lab to avoid shipping large volumes of
formalin which can be expensive and increase the risk of leaking
2 Tissue Fixation
6 Denoting Margins
7 Other Things to Avoid
8 What Else Should I know?
It is important for you to realize that after all is said and done
the pathologist typically evaluates 1 to 4, 5um thick sections
from the entire specimen which is submitted (A).
Our staff and pathologists are here to assist you. If you have
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!
1 Clinical Information
Please provide signalment and pertinent clinical information
Certain lesions occur more commonly in different species and
certain breeds
Anatomical location of a lesion, as well as clinical appearance and
progression, may also be critical information to allow your
pathologist to provide you with the best possible diagnosis and/or
differentials
If you have a specific question, are concerned about a possible
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
submission form. It will help us help you help your patients.
5 Endoscopic Biopsies
B. Large samples can be held
and fixed at your clinic prior to
submitting to the lab to help
avoid shipping large volumes of
formalin which may be costly and
hazardous
B
C. This is an example of an ~20cm
diameter mass lesion which was fixed
at the clinic and subsequently sent to
the lab in a plastic, labeled, zip lock
bag devoid of any formalin. (bar =
~2.5 cm)
C
A. Incomplete parallel cuts at a
minimum of 2cm apart (bread
loafing) can be utilized to assist
with appropriate tissue fixation
for large specimens
A
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
formalinTagging
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)
Tumor Bed Samples
1. Submission of samples from the post-surgical
bed 2. Any tumor in these
specimens is evidence of remaining microscopic
disease 3. Similar to
“submitting multiple sites” clearly label and submit
each region individually
Formalin filled jars containing specimens should be placed in
a plastic bag, box, or other container with absorbent material to
absorb any leakage (A)
Paperwork should be placed in a separate plastic bag to avoid
contact with formalin if leaking does occur. Such contact can
result in altered and illegible paperwork (B)
Your fresh sample size should never be larger than the most
narrow portion of the jar in which you are submitting it (C). If it
is, this will require cutting plastic jars or breaking glass jars
(undesirable) in order to retrieve the tissue which may become
altered in the process.
YES
The optimal method by which to submit
multiple lesions from a single animal is to
submit each specimen individually in its own
respective and appropriately labeled jar. This
should be reflected on the submitted
paperwork.
If multiple specimens are submitted in a
single container (which is less ideal) there
needs to be some method of tissue
identification (i.e. suture) to denote
specimens relative to respective anatomic
site. YES
suture
NO NO YES
Do not submit endoscopic
biopsies wrapped in gauze
sponges. Specimens may
become lost or may be crushed
during the attempted retrieval
process. It is better to submit
the specimen free floating in the
jar than with gauze or any other
material
The optimal method by which to
submit an endoscopic biopsy is to
place it in a screen cassette after
which the cassette should be placed in
an appropriately labeled formalin filled
jar. If individual cassettes are labeled
properly (sharpie or no2 pencil),
multiple cassettes can be place in one
jar.
Do not place endoscopic
biopsies on fragments of
cardboard. Specimens will
either float off or, if adhered,
tissue will slough off during
retrieval and/or may be
associated with cardboard
fibers
A. From
mass to
block to
slide
Our staff
is here
working
hard for
you!
Please help keep our technicians
fingers safe and DO NOT submit
specimens with needles for any
reason!
NO
NOPlease do no staple, suture, or pin
tissue to cardboard. It can damage
tissue and prevent appropriate margin
assessment
YES NO
YES
NO
A
CB
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
• Fiksatif Buffered Neutral Formalin 10%
• Volume Fiksatif : Organ = 10 : 1
• Waktu fiksatif yang cukup minimal 48 jam sebelum prosesing jaringan
Sampling Organ
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
JARINGAN
Contoh PengirimanSample 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. Deskriptif2. Skoring Lesio dengan metode Histoskor (Intensitas x
Distribusi), yang dilanjutkan dengan Analisis Statistika
Studi Infeksi Virus Parvovirus - MyokarditisIntranuclear inclusion body
Sarang radang
AnalisisHistopatologi
TahapanKerusakan
otot jantungakibat
Kardiotoksin
Studi HistopatologiKinetika Tumor
Basal Sel Tumor
TC
HP
StudiTransplantasiSel Tumor
Pada Nude Mouse
Studi Tumor Imunologi
ImunohistokimiaAntibodi-Antikeratin
• Signalemen
• Anamnese
• Temuan Patologi Anatomi
• Histopatologi
• Diagnosa/ Diferensial diagnosa
Penulisan Laporan
REFERENSI
• Henrik Elvang JensenDepartment of Veterinary Pathobiology, Royal Veterinary and Agricultural University, Copenhagen, Denmark
• Vincenzo Covelli - Guide to the Necropsy of the Mouse
• Dan lain lain
Terima Kasih
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