A 1 mg/mL (3.14 mM) Nile Red stock solution was prepared by dissolving 1 mg of Nile Red (ThermoFisher, N1142) in 1 mL of DMSO. A 1 mM Vybrant DiD stock solution (ThermoFisher, V22887) and a 20 mM Hoechst 33342 stock solution (ThermoFisher, 62249) were purchased directly. Cells were first stained in 2 µg/mL (6.28 µM) Nile Red in PSW for 20 min, then washed in PSW. Cells were then stained in 2.5 µM DiD and 20 µM Hoechst 33342 in PSW for 20 min, then washed in 20 µM Hoechst 33342 in PSW. Finally, cells in 20 µM Hoechst 33342 in PSW were loaded into the cage traps and imaged immediately. We found that Hoechst did not stain well under the conditions tested, so we opted to leave it in the solution during imaging to maximize the fluorescence signal.
Vybrant did stock solution
Manufactured by Thermo Fisher Scientific
Vybrant DiD stock solution is a fluorescent dye used for labeling lipid rafts and plasma membranes. It is a lipophilic carbocyanine dye that readily integrates into the lipid bilayer of cell membranes.
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Lab products found in correlation
2 protocols using vybrant did stock solution
1
Fluorescent Labeling of Cells
2
Multicolor Fluorescent Cell Staining
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A 1 mg/mL (3.14 mM) Nile Red stock solution was prepared by dissolving 1 mg of Nile Red (ThermoFisher, N1142) in 1 mL of DMSO. A 1 mM Vybrant DiD stock solution (ThermoFisher, V22887) and a 20 mM Hoechst 33342 stock solution (ThermoFisher, 62249) were purchased directly. Cells were first stained in 2 μg/mL (6.28 μM) Nile Red in PSW for 20 minutes, then washed in PSW. Cells were then stained in 2.5 μM DiD and 20 μM Hoechst 33342 in PSW for 20 minutes, then washed in 20 μM Hoechst 33342 in PSW. Finally, cells in 20 μM Hoechst 33342 in PSW were loaded into the cage traps and imaged immediately. We found that Hoechst did not stain well under the conditions tested, so we opted to leave it in the solution during imaging to maximize the fluorescence signal.
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