Observations on the living developing nerve fiber

The immediate object of the following experiments was to obtain a method by which the end of a growing nerve could be brought under direct observation while alive, in order that a correct conception might be had regarding what takes place as the fiber extends during embryonic development from the nerve center out to the periphery. The method employed was to isolate pieces of embryonic tissue known to give rise to nerve fibers, as for example, the whole or fragments of the medullary tube, or ectoderm from the branchial region, and to observe their further development. The pieces were taken from frog embryos about 3 mm. long, at which stage, i. e., shortly after the closure of the medullary folds, there is no visible differentiation of the nerve elements. After carefully dissecting it out the piece of tissue is removed by a fine pipette to a cover slip upon which is a drop of lymph freshly drawn from one of the lymph sacs of an adult frog. The lymph clots very quickly, holding the tissue in a fixed position. The cover slip is then inverted over a hollow slide and the rim sealed with paraffine. When reasonable aseptic precautions are taken, tissues will live under these conditions for a week and in some cases specimens have been kept alive for nearly four weeks. Such specimens may be readily observed from day to day under highly magnifying powers. While the cell aggregates, which make up the different organs and organ complexes of the embryo, do not undergo normal transformation in form, owing no doubt in part to the abnormal conditions of mechanical tension to which they are subjected, nevertheless the individual tissue elements do differentiate characteristically.