Objective To discuss injection methods that macromolecular tracer entered rat caudate-putamen (CPu) in order to find a better way to study the drainage pathway from the parenchyma of the brain. Methods Adult Sprague-Dawley rats were randomly divided into cutting needle group, non-cutting needle 4ul group and non-cutting needle 1μl group. Evans Blue-labeled albumin (EBA) solution was injected into the rat CPu with microsyringe under stereotaxic apparatus' control. Frozen sections of rat brain samples were made and observed by naked eye and fluorescence microscope respectively. Results The cutting needle way by which tracer's dose is injected into CPu was inaccurate and unstable, however, non-cutting needle way was accurate and stable. Conclusion Non cutting needle wethod by which tracers were injected into rat CPu was economical, accurate, stable and suitable for researching drainage of macromolecules from the parenchyma of the brain.