university of california, san diego histology shared resources

Dissection Protocol
Equipment and Materials:
Styrofoam board or cork cutting board and pins, flat for cutting
Scissors, delicate ½”—Fisher Cat. No. 08-951-5
Bell jar with lid
200ul Pipetman and pipets
Kimwipes, Lab liners, paper towels
Forceps, curved—VWR Cat. No. 25714-007
Secureline Marker—Fisher Cat. No. 14-905-30
1ml syringe—Fisher Cat. No.
Vacutainer 23G ¾ --Fisher Cat. No.0266539
HistoPrep Tissue Capsules—Fisher Cat. No. 15-182-219
Peristaltic Pump
Phosphate Buffered Saline
Isoflurane—MWI Cat. No. 012497
10% Neutral buffered formalin (Buffered Formaldefresh)--Fisher Cat. No. SF93-4
labeled specimen cups
Personal protective equipment eg. lab coat, gloves, eye protection.
Check out this website: www.eulep.org for photographs and other details
Before Autopsy:
1. Determine which organs will be harvested and formalin fixed or frozen.
2. Gather materials and label all necessary cassettes or capsules (see Blocking Chart Protocol.)
3. Record the identifying data on each animal including its appearance and behavior.
4. Record the weight of the animal, the individual organs, any abnormal gross pathology, and the
carcass after the organs are removed. (Relative organ weights can be found on page_____.) If
skeletal defects are suspected save the carcass and obtain a radiograph of the skeleton. The carcass
may be “cleared” using potassium hydroxide and stained with Alizarin Red and Alcian Blue,
following established protocols.
5. Anesthetize and euthanize animals humanely according to animal welfare guidelines.
6. Wet the animals' fur with 70% ethanol to minimize contamination with hair and lay upon a clean
paper towel. Pin extremities to styrofoam or cork board. Dedicated scissors are recommended for
cutting skin and bone.
7. Start from the abdomen and make and a Y-shaped incision should be made to open up the thorax
and abdomen. The ribcage should be opened bilaterally, exposing the underling thymus, heart and
lungs.
8. If the plan is to perfuse the entire animal, canulate the left ventricle of the heart with the catheter
connected to the peristaltic pump and secure the catheter with alligator clips..
9. Make an incision in the right atrium to allow the circulating fluids to exit the right atrium.
10. Pump PBS through the circulatory system, using a peristaltic pump. This will flush out blood.
Adequate perfusion is indicated when the liver and kidneys become pale.
11. Follow the PBS flush with freshly made 4% paraformaldehyde for 15 min.
12. Just before finishing the perfusion, insert a formalin filled syringe into the trachea just above the
lungs until full. Then tie off the trachea to keep the fixative in the alveoli
13. Once perfusion is complete, remove the organs listed below in the order listed. This will ensure
that all the necessary structures are collected.