After in situ perfusion with cold PBS and subsequently with cold buffered formalin, the arch and thoracic portion of the dorsal aorta are isolated from the arch to the aorticiliac bifurcation. After removing the adventitia and adipose tissue it is placed in 4% formalin overnight. The aorta was then opened lengthwise, and pinned flat. The tissue was stained for 15 minutes with 0.5% Sudan IV solution in acetone and 70% ethanol (1:1). The tissue was decolorized for 5 minutes using 80% ethanol, and then washed gently with water for several minutes.
Sudan IV stain | Working Solution |
---|---|
Sudan IV | 5 g |
70% ethanol | 500 ml |
100% acetone | 500 ml |
20% SDS | 5 ml |
dH20 | to 500 ml |
Reference long:20610861