Soft x-ray lithographic studies of interphase chromosomes.

Soft x-ray absorption lithograph patterns of purified interphase human nuclei and chromosome arrays, imaged on PMMA resists, were examined by scanning EM. The patterns obtained were compared to those utilizing more conventional sources, including transmission EM, scanning EM, high voltage EM, and various light microscopic techniques. The x-ray resist images revealed orderly arrays of absorption profiles in the 3-dimensional specimen with both mild and more extensive developments of the resist. Dense chromatin at the edge of interphase nuclei revealed aligned periodic peaks on the order of 2200 A diameter, with substructure. The periodicity and alignment of interphase chromosomes were entirely consistent with birefringent data on nuclei indicating a high degree of 3-dimensional order. This degree of 3-dimensional order was observed in nuclei containing essentially DNA and histones with only very few other minor (probably structural) proteins. Sonication and nuclease treatment to disperse interphase chromosomes revealed similar absorption periodicities in individual chromosome fibers. Analysis of x-ray absorption profiles thus appears to offer significant new insights into the ordered structure of these defined biological specimens.