India Ink Recipe

The day prior to perfusions, mix the following:

100 ml 0.2 M phosphate buffer
12 - 15 g gelatin (the amount depends upon the Bloom)
50 ml dH2O

store in refrigerator overnight

On the day of perfusions heat the solution above and add 50 ml India ink

Deeply Anesthetize rat
Perfuse first with 0.1M Phosphate buffer until blood is cleared using a rate of 35 ml/min* (see Davies et al., 1996). This should take about 1-2 minutes.

Then switch to warmed (37° C) India Ink solution. To keep the solution at the correct temperature, you may want to store the container in a warmed water bath during perfusions.

* The perfusion rate may change depending on the age of the animal and species. This rate is appropriate for adult rats, and was obtaine from Davies KR, Richardson G, Akmentin W, Acuff V, Fenstermacher JD (1996) The microarchitecture of cerebral vessels. In Couraud and Scherman (Eds) Biology and Physiology of the Blood-Brain Barrier. New York:Plenum Press.