Next-generation red ultra-bright fluorescent dyes for nuclear imaging and peripheral blood leukocytes sorting

The nucleus is a membrane-bound organelle in eukaryotic cells and plays a crucial role in cellular processes. Visualizing nuclear morphology is essential for investigating nuclear functions and understanding the relationship between nuclear morphological alterations and multiple diseases. Fluorescent dyes have been developed to visualize nuclear morphology, but the selection of red nuclear-labeling fluorescent dyes remains limited (high price, unknown structure, or high toxicity). Herein, we have developed a red ultra-bright nuclear-targeted dye, BPC1, through the engineering of unsymmetrical cyanine dyes derived from D-π-A systems. BPC1 exhibits ultrahigh fluorescence brightness and exceptional cell permeability, and selectively stains nuclear DNA rather than mitochondrial DNA, enabling the visualization of the nucleus in diverse cells at extremely low doses (100 nM) and laser power (0.8 μW). Furthermore, BPC1 is utilized for nuclear staining in blood cells, aiding in the distinct visualization of the white blood cell nucleus and facilitating the identification and enumeration of various leukocyte types. Our study implies considerable commercial potential for BPC1 and underscores its capacity to serve as a powerful tool in life sciences and cell biology research.