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Lipophilic Tracer Sampler Kit

Lipophilic carbocyanine dyes are cyanine fluorescent dyes bearing long hydrophobic hydrocarbon tails. These dyes exhibit low fluorescence in an aqueous environment but show high fluorescence when incorporated into lipid bilayers, which makes them valuable for staining membranes. They effectively and durably label cytoplasmic membranes and various intracellular membrane structures across a wide range of cell types ^[1]. These dyes have been employed as tracers in investigations involving cell fusion ^[2], cellular adhesion ^[3], and migration ^[4] due to their minimal toxicity and strong resistance to transfer between cells. By using different fluorescent colors, cell populations can be uniquely labeled for subsequent identification after mixing. Double labeling can help pinpoint cells that have fused or formed clusters ^[5]. Additionally, these dyes are widely used as anterograde and retrograde axonal tracers in living ^[1,6] and fixed ^[7,8] neuronal tissues and cells.

This kit contains samples of eight carbocyanine dyes with different membrane diffusion rates, solubility, and fluorescence properties:

• DiI (DiIC18(3); 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindocarbocyanine) is a bright and photostable orange-red fluorescent lipophilic dye widely used for membrane labeling, cell tracking, axonal tracing, and super-resolution imaging by STORM ^[9].
• DiO (DiOC18(3); 3,3’-dioctadecyloxacarbocyanine) is a widely used green fluorescent lipophilic dye with a slower lateral diffusion rate on the membranes than DiI. DiO is often used together with DiI in dual-color studies.
• DiA (4-Di-16-ASP; 4-(4-(dihexadecylamino)styryl)-N-methylpyridinium iodide) is a yellow-green fluorescent lipophilic dye which diffuses much faster than DiO in cell membranes. DiA is often used together with DiI for dual-color labeling.
• DiD (DiIC18(5); 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindodicarbocyanine) is an analog of DiI with red-shifted fluorescence excitation and emission spectra allowing to avoid autofluorescence and phototoxic effects during imaging. The dye is used for multicolor labeling and super-resolution imaging by STORM ^[9].
• DiR (DiIC18(7); 1,1’-dioctadecyl-3,3,3’,3’-tetramethylindotricarbocyanine) is a near-infrared (NIR) fluorescent lipophilic dye. The NIR region of the spectrum is optically transparent for many tissues so that DiR can be used for small animal in vivo imaging ^[10] and super-resolution microscopy by STORM ^[9].
• Neuro-DiO is a green fluorescent lipophilic carbocyanine dye. In contrast to DiO, Neuro-DiO has been specifically designed to improve solubility in membranes and prevent the formation of nonfluorescent aggregates, which tend to decrease the dye diffusion rate.
• RAPID DiI (dilinoleyl DiI; FAST DiI™; 1,1’-dilinoleyl-3,3,3’,3’-tetramethylindocarbocynanine) is an unsaturated analog of DiI with faster lateral diffusion rate in cell membranes than original DiI. It is especially useful for tracing neurons both in live and fixed tissues.
• RAPID DiO (dilinoleyl DiO; FAST DiO™; 1,1’-dilinoleyl-3,3’-oxacarbocynanine) is an unsaturated analog of DiO with a faster lateral diffusion rate in cell membranes than the original DiO. It is especially useful for tracing neurons both in live and fixed tissues.

References

^[1] J Cell Biol 103, 171 (1986); ^[2] Cytometry 21, 160 (1995); ^[3] J Cell Biol 136, 1109 (1997); ^[4] Anticancer Res 18, 4181 (1998); ^[5] J Immunol Methods 156, 179 (1992); ^[6] Trends Neurosci 12, 333 (1989); ^[7] Methods Cell Biol 38, 325 (1993); ^[8] Development 101, 697 (1987); ^[9] Proc Natl Acad Sci U S A 109, 13978 (2012); ^[10] Curr Drug Deliv 13, 40 (2016).