We need to purge the kidneys of intravascular blood cells prior to leukocyte quantification. Thanks in advance.
It is a good idea to perfuse while the animal is still alive, otherwise the renal vasculature constricts. Look at the literature on perfused rat kidney - a short 90° glass cannula is the usual tool. This reference may help get you started: Adam WR, Adams BA.
Ren Physiol. 1985;8(3):150-8.
retrograde by characterization of the abdominal aorta. First catheterize below the renal arteries using a clip to occlude blood flow and then ligate the aorta superior to the renal arteries after the perfusion has started. You will also need to cut the vena cava to prevent back pressure. Its also a good idea to ligate the celiac and mesenteric arteries to maintain perfusion of the kidney with PBS. We use 4mL/min per kidney.
You can also try to directly perfuse through the renal vein - refer to our hydrodynamic renal vein delivery method
Article A Method to Facilitate and Monitor Expression of Exogenous G...
Paul M O'Connor's answer is the definite answer for pressure fixation (e.g. with PFA). The only different way to go would be to monitor not flow, but pressure during the perfusion. We like to use high pressure in the beginning, which we then lower. 200mmHg for 5 minutes, then 100mmHg for another 5 minutes. It is useful and gives better results if the perfusion solution is not ice-cold but warmed to 37°c at this time point. Additionally, it makes sense to first flush the kidney with 5ml of 2% Na-Heparin added into the buffer.
If you just want to clear the kidneys of intravascular blood, though, and not fix them, perfusing PBS through the left ventricle is the option for rapid perfusion of many animals. The upside to this method is that it can be mastered quickly also by the technically unadvanced person. You can open the right ventricle for releasing blood and perfuse through the left ventricle, which is easiest hit puncturing the tip of the heart.
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