AIM:To observe the absorption of tetrandrine sustained-release absorption film in vivo in rabbit conjunctiva under the laser confocal microscopy. METHODS:Tetrandrine film prepared with an appropriate proportion of chitosan and gelatin was implanted under the conjunctiva of rabbits,blank film as control.1 week later the two groups of film was observed with the confocal microscope at five different levels.The average percentage of absorption area of the film was calculated. The difference of absorption was analyzed by the SPSS 18.0. RESULTS:Lots of uniform small particles in the dry film was observed under the laser confocal microscope.At the same time,there were some vacuoles and the absorption regions in the film under the rabbit conjunctiva.The average absorption percentage was 10.5%±0.28%in the tetrandrine film group,and 15.0%±4.6%in the control. Tetrandrine film absorption in the rabbits conjunctiva was lower,with t= 1.301 and P= 0.000. CONCLUSION:The laser confocal microscope can be used to observe the degradation of film directly,and to get the result of the tetrandrine releasing the biological absorption of the film.So it is a good tool of the pharmaceutical research.