Live Imaging of Ependymal Cell Ciliary Activity in a Mouse Brain Section

Take a sagittal section of a mouse brain to expose the ventricles, which are cavities filled with cerebrospinal fluid, or CSF.

The ependymal cells lining the ventricles possess motile cilia that propel CSF to facilitate its circulation.

Place the section in a glass-bottom dish containing a nutrient-rich medium to sustain ciliary activity.

Transfer the dish to the climate-controlled chamber of a differential interference contrast, or DIC, microscope to maintain cell viability during imaging.

DIC microscopy employs a prism to split polarized light, directing it through the specimen along different paths.

Variations in refractive index across the specimen introduce phase differences between the beams.

A second prism recombines the beams, creating interference that produces a high-contrast image with a three-dimensional effect, enhancing the visibility of fine structures.

Locate functional ependymal cells with motile cilia by observing bubble movement generated by their beating.

Using high-contrast DIC imaging, record the cilia to quantify their beating frequency.