Light and electron microscopy of laser microirradiated nucleoli and nucleoplasm in tissue culture cells

This manuscript describes in precise detail the ultrastructural alterations produced as a result of laser microirradiation of nucleoli and nucleoplasm of tissue culture cells. Because of the general difficulty of single cell recovery, flat embedding, and serial sectioning, very few studies have ever been conducted on microbeam irradiated cells; yet the use of the microbeam technique has become widespread in functional studies of the nucleus. The results presented here demonstrate two classes of lesion material: small spherical electron dense bodies 0.05-0.02 μm in diameter and a larger, more irregular electron dense material up to 1 μm in length. The occurrence of these different types of lesion materials is described in control irradiated nucleoli and nucleoplasm, irradiated nucleoli and nucleoplasm in quinacrine treated cells, irradiated nucleoli and nucleoplasm in actinomycin D treated cells, and irradiated nucleoli and nucleoplasm in combined actinomycin D and quinacrine treated cells. In all the cells in which actinomycin D was employed, nucleoli were selectively irradiated in either their granular or fibrillar zones. The results of the ultrastructural studies are discussed in light of earlier functional studies.