The use of scanning electron microscopy for investigations into the three dimensional organisation of the interphase nucleus.

In an attempt to elucidate three dimensional information on the organisation of the nucleus, nuclei have been extracted from ascites tumour cells or tissue culture cells by a variety of biochemical techniques, and prepared for high resolution scanning electron microscopy using an osmium-thiocarbo-hydrazide infiltration procedure which has previously proved successful for analysis of chromosome structure. Nuclei were prefixed with either Methanol-Acetic acid, glutaraldehyde or formaldehyde and then extracted by a variety of detergents with the aim of a 'biochemical dissection' of their outer elements to allow surface visualisation of the nuclear lamina. Continued extraction removed all aspects of the nuclear periphery and allowed direct visualisation of the in situ organisation of the chromatin, apparent as at least two levels of supercoiling.