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PKH Fluorescent Cell Linker Kits permit stable, reproducible, fluorescent cell labeling. Patented cell linker technology incorporates aliphatic reporter molecules into the cell membrane lipid bilayer by selective partitioning.¹ Three fluorescent linkers are available. PKH2 and PKH67 are green fluorochromes with excitation (490 nm) and emission (504 nm) similar to fluorescein, while PKH26, a red fluorochrome, has excitation (551 nm) and emission (567 nm) characteristics compatible with rhodamine or phycoerythrin detection systems. PKH26 may also be excited by the 488 nm emission of an argon-ion laser. The linkers are physiologically stable and show little to no toxic side-effects on cell systems. Labeled cells retain both biological and proliferative activity, and are ideal for cell tracking and cell-cell interaction studies. Due to the non-specific labeling mechanism of the cell linkers, a wide variety of cell types have been labeled successfully. The linkers have been applied to both animal and plant cells as well as other membrane containing particles. The pattern of staining is dependent upon the cell type being labeled and the membrane of the cells.² Although most applications center around general labeling (GL) methods involving membrane incorporation of the probes, the linkers may also be used for selective phagocytic cell labeling (PCL). Appearance of labeled cells may vary from bright "immunofluorescence" labeling to a punctate or patchy appearance. Since the labeling is not a saturation reaction, but rather a function of both dye and cell concentration, it is essential that the amount of dye available for incorporation be limited. Overlabeling of the cells will result in loss of membrane integrity and cell recovery.

References

1. Horan, P., and Slezak, S., Nature, 340, 167 (1989).

2.Horan, P., et al., Meth. Cell Biol., 33, 469 (1990).

General Reference:

Horan, P., et al.,"'Fluorescent Cell Labeling for in vivo and in vitro Cell Tracking," in Methods in Cell Biology, Vol. 33, Flow Cytometry, (eds., Darzynkiewicz, Z., and Crissman, A.), Academic Press, Inc., page 469 (1990).