We describe a method for labeling cultured endothelial cells (ECs) and smooth muscle cells (SMCs) by letting the cells grow for three days in culture medium containing a low concentration of the fluorescent carbocyanine dyes DiI and DiO. We show that good labeling can be obtained with considerably lower concentrations (2.5 micrograms/ml) than has previously been described. With optimal concentration the labeling is very strong and seems to label all membranous structures in the cells. It was possible to clearly distinguish differentially pre-labeled cells both in coculture and seeded on denaturated vascular grafts. The cells remain fluorescent for more than seven days and may be passaged with retained proliferative capability. We suggest that DiI/DiO-labeling using dye-containing medium may be used for several cell types and is applicable in tissue culture and in the detection of implanted cells in vivo.
We have examined the topographic relationship between the sagittal bands of zebrin I immunoreactive Purkinje cells revealed by a monoclonal antibody, mabQ113, and the distribution of spinocerebellar fibers originating from the central cervical nucleus in the rat. The mossy fiber terminals were anterogradely labeled following injections of cholera toxin subunit B into the C1-C3 segments and visualized immunohistochemically. Zebrin I positive Purkinje cells appeared in seven sagittal bands (P1+ to P7+ bands). In lobules I-V of the anterior lobe, labeled mossy fiber terminals were distributed in the midline region, subjacent to the P1+ bands and at around 0.5 mm from the midline region, subjacent to the P2+ band in the lateral A1 to the medial A2 zones of Voogd et al. (1985). Labeled terminals were seen in the entire B zone and those distributed in its medial part were related to the P3+ band. In lobule VIII, labeled terminals were seen subjacent to the P1+, P2+ and P3+ bands, which were located in the lateral A1-A3 (or B) zones. In the copula pyramidis, labeled terminals appeared subjacent to the P4+, P5+ and the P6+ bands in the C1 and C2 zones (or the C1-C3 zones). Although the labeled terminals were seen beneath the zebrin I positive bands, the borders of terminal distribution were not well-delineated, and did not respect the borders of zebrin I positive bands.
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