[Three-dimensional visualization of the brain ependyma and tanycytes]

The aim of this investigation was to develop an integrated approach to spatial reconstruction of the cells lining the ventricles of the brain using confocal laser microscopy and immunocytochemical reaction to vimentin. The work was performed on paraffin sections of rat brain of different thickness (5 and 10 microm). To visualize the immunocytochemical reaction the fluorescent dyes in the visible range were selected: SYTOX Green selectively staining the nucleus and indocarbocyanin (Cy-3) conjugated with streptavidin. As a result of testing of various processing conditions, the protocol which allows to receive an intensive staining of the structures was developed. The set of fluorochromes proposed in confocal laser microscopy allows to separate easily the channels, to study the structures independently, if needed, and does not require the use of an expensive ultraviolet laser.