The animals were anesthetized by intraperitoneal injection of zoletil (12.5 mg) and xylazine (3 mg), and fixative containing 4% buffered paraformaldehyde was perfused through the left ventricle.
Isotonic phosphate buffer saline (IPBS) (pH 7·4) was perfused through the probe with a flow rate of 2 μl ml−1.
To measure protein concentration, 50 μL Tris-buffered saline containing NP40 (1% vol/vol) was perfused through the capillaries, the lysates were collected, and Bradford Protein Assay was performed.
Using this pump a modified Krebs-Henseleit buffer, with the addition of a colloid (40 g/L dextran-70), was perfused through the microdialysis catheter at a perfusion rate of 0.3 μL/min.
The liver was perfused through the portal vein with buffer I [D-Hanks (Invitrogen, Grand Island, NY, USA) media containing 0.05% collagenase IV (Sigma-Aldrich), 0.1% pronase E (Roche, Madison, WI, USA)] at 37°C for 20 min.
At the end of the experiment, the right kidney was removed and quickly frozen for molecular studies, and the left kidney was perfused through the femoral catheter with phosphate buffered saline (PBS).
The myocardium was perfused through the aorta at a constant pressure of 80 mmHg using 0.1 M cacodylate buffer (3 min) followed by 2.5% glutaraldehyde solution diluted in cacodilate buffer.
Briefly, whole blood loaded with 3,3′-dihexyloxacarbocyanine iodide was perfused through collagen coated (100 μg/mL) glass micro-capillaries (shear rate = 1000 s−1), followed by Tyrodes–Hepes buffer.
Each rat was perfused through the heart with 50 ml 1% sodium nitrate, followed by 500 ml 4% paraformaldehyde in 0.1 M phosphate buffer (PB).
