Fluorescent probes of chromosome structure and replication.

Procedures employing fluorescent dyes or Giemsa stain have been utilized to differentiate methaphase chromosomes into longitudinal segments termed bands. In spite of the immense practical utility of chromosome banding, the chemical basis of banding patterns remains incompletely understood. Physical chemical studies have elucidated the modes and specificities of the interaction of fluorescent dyes such as quinacrine, 33258 Hoechst, daunomycin, chromomycin A3 and 7-aminoactinomycin D with DNA and chromatin. However, it is not clear that all aspects of chromosome staining are explainable in terms of the optical properties of soluble dye-DNA complexes. BrdU-dye techniques in which chromosome staining depends on the schedule of BrdU incorporation by cells, have been used for cytological studies of chromosome structure and replication. These procedures have revealed a close association between quinacrine or Giemsa bands and late replicating chromosomal regions. Biochemical studies on chromatin differentially labelled according to replication timing may thus prove useful for investigating the molecular basis of chromosome banding.