Characteristics of a novel deep red/infrared fluorescent cell-permeant DNA probe, DRAQ5, in intact human cells analyzed by flow cytometry, confocal and multiphoton microscopy.

摘要: Background The multiparameter fluorometric analysis of intact and fixed cells often requires the use a nuclear DNA discrimination signal with spectral separation from visible range fluorochromes. We have developed novel deep red fluorescing bisalkylaminoanthraquinone, DRAQ5 (Exλmax 646 nm; Emλmax 681 Emλrange 665–>800 nm), high affinity for capacity to enter living cells. describe here characteristics applications this synthetic compound, particularly in relation cytometric cell cycle. Methods Cultured human tumor were examined ability locate using single multiphoton laser scanning microscopy (LSM) flow cytometry. Results Multiparameter cytometry shows that dye can rapidly report cellular content live at resolution level adequate cycle cycle-specific expression proteins (e.g., cyclin B1). The preferential excitation by lines (633/647 nm) was found offer means fluorescence selective excitation, greatly reduced emission overlap UV-excitable fluophors as compared propidium iodide. LSM reveals architecture clearly defines chromosomal elements permit imaging nuclei 1,047-nm emitting mode-locked YLF laser. unusual properties also conventional 488 nm single-photon between low infrared (IR; 780 wavelengths. Single studies revealed three-dimensional structure location expressing endoplasmic reticulum targeted-GFP, MitoTracker-stained mitochondria, or vital probe free zinc (Zinquin). Conclusion The incorporation measurement into variety analyses. Cytometry 40:280–291, 2000 © Wiley-Liss, Inc.