Figure 8

The labeling and tracking of differentiated neuron cells using FNDs.

(a) A model of differentiated neuron cells labeled with FNDs. (b) P19 cells and neuron cells were treated with or without FNDs (50 μg/ml for 24 h). The fluorescence intensity of FNDs in P19 cells and neuron cells were measured by flow cytometer. The fluorescence intensity of FNDs was excited with wavelength 488 nm and the emission was collected in > 650 nm signal range (FL3-H). (c) The fluorescence intensity of FNDs was quantified from a minimum of 10,000 cells by CellQuest software. Results were obtained from three separate experiments and the bar represented the mean ± S.E. **p < 0.01 indicates significant difference between the ECS cells and neuron cells by treatment with FND, or the neuron cells treated with or without FNDs. ***p < 0.001 indicates significant difference between the untreated and FND-treated ECS cells. (d) P19 cells were differentiated into neuron cells by RA induction. Then the neuron cells were treated with FNDs (50 μg/ml for 24 h). At the end of treatment, the cells were incubated with mouse anti-SSEA-1 and rabbit anti-β-III-tubulin antibodies and then incubated with goat anti-mouse IgG Cy3 and goat anti-rabbit IgG HiLyte Fluor 488 antibodies. The nuclei were incubated with Hoechst 33258. The red fluorescence intensity of FNDs was excited with wavelength 580 nm and the emission was collected in 600–700 nm. The blue fluorescence intensity of nuclei was excited with 405 nm and the emission was collected in 415–485 nm. The green fluorescence intensity of β-III-tubulin was excited with 488 nm and the emission was collected in 505–545 nm. The fluorescence intensity of SSEA-1 was excited with 543 nm and the emission was collected in 560–580 nm.