Tubulin immunohistochemistry. Fixation methods affect the response of spinal cord cells in vitro

We report here the results observed when tubulin fluorescence immunohistochemistry is performed upon dissociated cultures of nervous tissue, principally of chick embryo spinal cord. When fixation includes nonpolar solvents or detergents, a uniform fluorescence is seen in neuron perikarya (with the exception of their nuclei), and the processes to which they give rise. Fixation with formaldehyde or glutaraldehyde alone, however, results in a discontinuous staining of neurites, and a less regular staining of their perikarya. The former pattern of response can be elicited if aldehyde fixation is followed by exposure to non-polar solvents. Such results are obtained both in thinly spread regions of the cultures, where neurons and their processes can easily be seen, and in the cell aggregates that also characterise them. Possible interpretations of these results are discussed.