Vancouver Rat Project Necropsy SOP 2012
DETAILED NECROPSY AND TISSUE COLLECTION PROCEDURE
1. Remove rats from freezer at 4:30 PM the previous day. Remove oropharyngeal swabs,
blood tube and serum from bag and return to freezer. Place packaged rats in the
necropsy fridge and allow to thaw overnight.
2. Record rat ID numbers and label whirl pack bags for the next day’s necropsies.
3. Collect all necessary equipment and containers. Label container jars with Rat ID
4. Prepare ectoparasitology area (outside biosafety cabinet), necropsy area (inside
biosafety cabinet), and photography area (outside biosafety cabinet).
5. Remove rat from packaging in ectoparasitology area. Suspend over plastic bowl and
brush vigorously with both the flea comb and brush. Transfer ectoparasites from bowl
to whirl pack using paint brush to move ectoparasites if necessary.
6. Place rat in photography area and photograph in right lateral recumbancy (remember to
include label with rat number in the photograph). Straighten rat’s spine before photo is
taken to accurately estimate length.
7. Place rat in necropsy area.
8. Open via full ventral incision (chin to anus) and make note of mammary gland
development for females. From this point forward any abnormalities should be
photographed. Photographs should always include the rat ID label. Tissues
demonstrating pathology should be collected for histopathology and culture/freezing.
9. Skin rat except leave one ear attached to skull. Collect a skin sample from lateral thorax
for Histopath. Collect rest of pelt for BB Skin.
10. Mammary Tissue: Note nipple size and development of mammary tissue. Sample
mammary tissue for histopath.
11. Skeletal Muscle Sampling
o Collect muscle from the gluteus (including sciatic nerve), quadracepts, triceps
(including the axillary lymph node), and diaphragm for both histopath and BB
Muscle.
12. Bones:
o Femur: Collect a femur for BB Bone and Histopath (split diagonally before placing
in formalin).
o Ribs (3 from mid-thorax): Collect for BB Bone and Histopath
13. Remove abdominal wall and ventral rib-cage. Perform basic internal exam prior to
removing any organs.
14. Remove and examine the pluck
o Tongue Tip: Collect the tip for BB Muscle.
Vancouver Rat Project Necropsy SOP 2012
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Remaining Tongue, Cranial Trachea and Thyroids: Collect for Histopath
Thymus: Remove from base of heart and ventral to trachea for Histopath
Caudal Trachea: BB Lung/Trachea
Lung: Separate the cranial and caudal lung lobes. Five samples that contain a
section of cranial and caudal lung should be sectioned for BB Lung/Trachea, WT
Lung, DNA Lung, RNA Lung, and Histopath
15. Heart: Section heart perpendicular to long axis ventral to the level of the AV valves.
Collect apex for BB Heart and base for Histopath.
16. Remove liver, stomach, duodenum, pancreas, and spleen. Examine liver.
17. Liver: Collect one sample of liver for each of DNA Liver, RNA Liver, WT Liver (slightly
larger piece), and histopath (4 samples total). Collect the remaining liver tissue for the
BB Liver.
18. Spleen: Divide the spleen into 4 sections. Make sure the DNA Spleen and RNA Spleen
have large enough samples. BB and Histo can be small pieces. Collect 1 section for each
of DNA Spleen, RNA Spleen, Histopath, and BB Spleen. Note: for very small spleens,
make sure to have enough for DNA, RNA. Then if there is enough, for Histo, finally, if
there is enough, BB.
19. Kidneys: Remove and examine the kidneys and adrenal glands. Divide the kidneys into 5
sections.
o 1 Adrenal and 1 Kidney Section: BB Kidney/Ad/BL, and Histopath
o Collect one section of kidney for each of DNA Kidney, RNA Kidney, and WT
Kidney.
o Bladder: Remove urine from bladder using syringe and collect for biobank.
Divide the bladder into 2 sections and collect 1 for BB Kidney/Ad/BL and 1 for
histopathology. Note small rats: collect whole bladder for Histo
20. Reproductive Organs: Examine and photograph (if mature female). Make note of
reproductive status. Count and describe placental scars or fetuses.
21. Brain: Remove the skull cap. Section the brain in ½ along the the long axis and collect ½
for BB Brain. Leave the remaining ½ brain in skill and collect entire head for Histopath
22. Remove and examine the intestinal tract.
o Duodenum & Pancreas: Collect for Histopath
o Jejunum: Collect 3 (>2 cm long) samples at different levels for Histopath
o Ileum: Collect 1 sample for Histopath
o Feces: Collect feces, colon, cecal, or ileum contents as necessary for C. diff feces,
Bact Feces, DNA Feces, and RNA Feces. C.diff and Bact need slightly larger
samples than DNA/RNA. Keep within GI tract (ie. Do not express out) If there is
minimal contents try to include a sample from different parts for each.
Vancouver Rat Project Necropsy SOP 2012
o Colon: Collect a sample for Histopath
o Remaining Gastrointestinal tract: Collect for BB GI
23. Open stomach to examine contents.
o Stomach: Collect stomach contents for BB Stomach. Collect a longitudinal
section of stomach wall (cardia to pylorus) for Histopath
o Collect all reproductive organs for histopathology.
COLOR CODING AND LABELS
WHIRL PACK LABELS
Use only Sharpie Markers!
Rat ID number should be written in the upper 2/3 of bag. Make writing large.
Whirl Packs should be kept roughly in this order:
Ecto
BB Skin
BB Muscle
BB Bone
BB Heart
BB Lung/Trachea
DNA Lung
RNA Lung
WT Lung
BB Kidney/Ad/Bl
DNA Kidney
RNA Kidney
WT Kidney
BB Spleen
DNA Spleen
RNA Spleen
BB Liver
DNA Liver
RNA Liver
WT Liver
Vancouver Rat Project Necropsy SOP 2012
C. diff Feces
Bact Feces
DNA Feces
RNA Feces
BB GI
BB Stomach
BB Brain