Quantification of axonally transported material using cytofluorimetric scanning

This paper describes in detail a cytofluorimetric scanning technique used for studying amounts of material axonally transported in antero- and retrograde direction in peripheral nerves. Operating procedures, preparation of tissues and instrumental set-up are described. The basis for quantification of material in a nerve section treated for immunofluorescence is discussed. The reliability of the method has been tested by comparing results with biochemical data. There are several advantages of the technique. (1) Many different substances can be studied in one single nerve segment, thus reducing biological variation and costs. (2) Both morphological data and quantitative figures can be obtained; following scanning the section can be photographed. (3) The method can also be used on studies in the central nervous system and on tissue cultures, since it is possible to scan on single axons or bundles of fibres.