Time-Lapse Imaging of Mouse Neural Stem Cell Division Using Confocal Microscopy

Take a multi-well plate containing adhered neural stem cells or NSCs isolated from a transgenic mouse brain. These NSCs contain a fluorescently tagged protein that is specific to the first growth, or G1, phase.

As the NSC divides, it fluoresces in the G1 phase of the cell cycle while emitting no fluorescence in other phases.

Place the plate in a confocal laser scanning microscope imaging chamber, maintained at a controlled temperature.

Select the desired objective lens and adjust the fluorescence intensity.

Configure the parameters for time-lapse imaging.

Choose the large image option and schedule image acquisition at regular intervals for the desired duration.

Use a high-speed resonant scanner to avoid photobleaching and maintain high resolution.

Use the brightfield setting to visualize the cell shape, and use an appropriate fluorescent filter to capture the fluorescence.

Start time-lapse imaging of the NSC to visualize different cell cycle phases.

Prepare a confocal laser scanning microscope for imaging by preheating the attached temperature control chamber to 37 degrees Celsius under a 5% carbon dioxide atmosphere. Position the 96-well plate inside the prewarmed microscope chamber and b