Source: Perumal, N. et al., Sample Preparation for Mass-spectrometry-based Proteomics Analysis of Ocular Microvessels. J. Vis. Exp. (2019).
This video describes the protocol to isolate the short posterior ciliary arteries from the porcine eye. The isolated short posterior arteries are a suitable source for elucidating the molecular insights and protein profiles of the ocular blood vessels.
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. Isolation of Short Posterior Ciliary Arteries
Porcine eyes together with optic nerve and extraocular tissues were obtained immediately post-mortem. Enucleated eyes were transported to the laboratory in ice-cold phosphate buffered saline (PBS) and used immediately.
NOTE: The porcine eye is basically divided into the anterior (Figure 1A) and posterior sections (Figure 1B).
2. Sample Preparation
Figure 1: Representative photographs of the porcine eyes.
(A) Lateral view of the eye globe shows the cornea, which is located in the anterior part of the eye, the sclera, surrounding muscle and optic nerve. (B) Posterior view of the eye shows the optic nerve. (C) Branches of sPCA seen at the back of the eye globe composed of the paraoptic and distal branches. (D) Isolated sPCA with surrounding fat and connective tissues.
The authors have nothing to disclose.
Calcium chloride dihydrate (CaCl2) | Carl Roth | 5239.1 | 2.5 mM |
Dulbecco's phosphate-buffered saline (PBS) | Thermo Fisher Scientific | 14190169 | |
Sodium chloride (NaCl) | Carl Roth | 9265.2 | 118.3 mM |
Sodium hydrogen carbonate (NaHCO3) | Carl Roth | 965.3 | 25 mM |
α-(D)-(+)- Glucose monohydrate | Carl Roth | 6780.1 | 11 mM |
Disposable scalpels #21 | pfm Medical | 200130021 | |
Dissection pins | Carl Roth | PK47.1 | |
Extra Fine Bonn Scissors | Fine Science Tools | 14084-08 | |
Falcon conical centrifuge tubes (50 mL) | Fisher Scientific | 14-432-22 | |
Mayo scissors, Tough cut | Fine Science Tools | 14130-17 | |
Precision tweezers | Fine Science Tools | 11251-10 | Type 5 |
Precision tweezers, straight with extra fine tips | Carl Roth | LH53.1 | Type 5 |
Standard pattern forceps | Fine Science Tools | 11000-12 | |
Student Vannas spring scissors | Fine Science Tools | 91501-09 | |
Vannas capsulotomy scissors | Geuder | 19760 | Straight, 77 mm |