[In vitro observation of ciliary activity of the tracheal membrane under a common light microscope].

ABSTRACT

OBJECTIVE: To establish a new method for in vitro observation of ciliary activity of the tracheal membrane in rabbit under a common light microscope. METHODS: Nine healthy adult rabbits were used. Two equal sized cervical trachea flaps were removed after the rabbits were anesthetized by urethane (4 mg/kg). The removed trachea flap was randomly assigned to the ephedrine (Eph) and DMEM control group, respectively. The observed trachea flap was placed in a small flat-bottomed glass container with its membranous side upward in DMEM culture medium solution (DMEM control group) or in 0.5% ephedrine solution prepared with DMEM (Eph group). One drop of 1% methylthioninium stained autologous blood cells was added into the glass container as the tracer, and the trachea flap was observed under a common light microscope (400 x). The latter was attached with a digital camera linking to an image manipulation system, the computed dynamic image analyser. The velocity of the tracer cell movement worked as the indicator for ciliary activity and was automatically determined by the digital image manipulation system. The variation coefficient (VC) was used as the indicator of the cell movement velocity. RESULTS: There was no significant difference among the VC at different time points in the DMEM control group. VC of the Eph group decreased regularly with the time point. CONCLUSION: A thin layer of flowing fluid was found on the surface of the tracheal mucociliary blanket which is driven by the activity of the mucociliary system. The new method of using the tracer to evaluate the ciliary activity of mammalia tracheal membrane in vitro is reliable and stable. It is practical and valuable in the in vitro observation and evaluation of ciliary activity of the tracheal membrane.