Cell Navigator® Live Cell RNA Imaging Kit *Green Fluorescence*

Protocol summary

1. Prepare cells
2. Add StrandBrite™ RNA Green working solution
3. Incubate for 30 - 60 minutes
4. Analyze the cells under fluorescence microscope at Ex/Em = 490/520 nm (FITC filter set)

Important notes
Thaw all the components at room temperature before starting the experiment.

1. Culture cells to a density optimum for imaging according to your specific induction protocol (about 1 - 2 × 10⁴ cells/well/96-well plate).
   
   
2. For living cells: Incubate cells with StrandBrite™ RNA Green (Component A) diluted 400X in medium or live cell staining buffer (Component B) at room temperature for 30 - 60 minutes (100 µL/well). Note: 25 µL of StrandBrite™ RNA Green (Component A) is enough for one 96-well plate. Protect from light and avoid repeated freeze-thaw cycles. The appropriate incubation time depends on the individual cell type and cell concentration used. Optimize the incubation time for each experiment. See figure 1 for details.
   
   
3. For fixed cells: Fix cells with pure methanol for 1 minute at room temperature, then wash with PBS. Immerse cells in 1% Triton-100 for 2 minutes, then wash with PBS twice. Incubate cells with StrandBrite™ RNA Green (Component A) at the concentration of 1X in PBS at room temperature for 15 - 30 minutes. Note: It is recommended to increase either the labeling concentration or the incubation time to allow the dye to accumulate if the cells do not appear to be sufficiently stained. See figure 1 for details.
   
   
4. (Optional) Wash the cells with PBS for 1 - 2 times, add 100 µL PBS to each well.
   
   
5. Monitor fluorescence intensity with fluorescence microscope at Ex/Em = 490/520 nm (FITC channel).