Animal experiment:

1 to 5×105 cells per mL HSPCs are stained with 5 μM DiD perchlorate (DiD) or 7.5 μM DiI in PBS without serum for 10 min at 37℃, washed once in PBS and immediately injected into the tail vein of recipient mice. Unless stated differently, each imaged CD45.2 mouse receive 8,000 to 15,000 labelled CD45.1 HSPCs together with 3 to 5×105 CD45.2 supportive whole bone-marrow mononuclear cells to ensure survival[1].

DiD perchlorate is a lipophilic cyanine dye.

Two weeks after injection of stained cells, a single, bright, DiD perchlorate (DiD)-positive cells located between 15 to 40 µm from the endosteum is found. Results reveal a progressive appearance of cell clusters of decreased dye intensity, consistent with the partitioning of DiD perchlorate label on cell division[1].

[1]. Lo Celso C, et al. Live-animal tracking of individual haematopoietic stem/progenitor cells in their niche.