carbocyanine-dye-diic12(3) and 3-3--dioctadecylindocarbocyanine

A non-ideal lipid binary mixture (dilauroylphosphatidylcholine/distearoylphosphatidylcholine), which exhibits gel/fluid phase coexistence for wide temperature and composition ranges, was studied using photophysical techniques, namely fluorescence anisotropy, lifetime and resonance energy transfer (FRET) measurements. The FRET donor, N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-dilauroylphosphatidylethanol amine, and a short-tailed FRET acceptor, 1,1'-didodecil-3,3,3',3'-tetramethylindocarbocyanine (DiIC12(3)), were shown to prefer the fluid phase by both intrinsic anisotropy, lifetime and FRET measurements, in agreement with published reports. The other studied FRET acceptor, long-tailed probe 1,1'-dioctadecil-3,3,3',3'-tetramethylindocarbocyanine (DiIC18(3)), is usually reported in the literature as partitioning mainly to the gel. While intrinsic lifetime studies indeed indicated preferential partition of DiIC18(3) into a rigidified environment, FRET analysis pointed to an increased donor-acceptor proximity as a consequence of phase separation. These apparently conflicting results were rationalized on the basis of segregation of DiIC18(3) to the gel/fluid interphase. In order to fluid-located donors sense these interphase-located acceptors, fluid domains should be small (not exceed approximately 10-15 nm). It is concluded that membrane probes which apparently prefer the gel phase may indeed show a non-random distribution in this medium, and tend to locate in an environment which simultaneously leads to less strict packing constraints and to favorable hydrophobic matching interactions.

Topics: 4-Chloro-7-nitrobenzofurazan; Carbocyanines; Fluorescent Dyes; Phosphatidylcholines; Phosphatidylethanolamines; Spectrometry, Fluorescence; Temperature

The carbocyanine dye DiIC18(3) ("DiI") is commonly used for both anterograde and retrograde labeling of neurons, including live neurons in situ and in vitro. In the present experiments, DiIC18(3) was used to label motoneurons in the spinal cords and sensory neurons in the dorsal root ganglia of embryonic rats. When the neurons from these regions were placed in culture, the neurons labeled by the dye were found to die rapidly, suggesting that DiIC18(3) can be toxic to neurons of these types. A related dye, DiIC12(3), was found to be equally suitable for labeling these neurons, and was found not to have detectable toxic effects in vitro.

Topics: Affinity Labels; Animals; Carbocyanines; Cell Survival; Cells, Cultured; Embryo, Mammalian; Flow Cytometry; Fluorescent Dyes; Kinetics; Motor Neurons; Neurons, Afferent; Rats; Rats, Inbred Strains