Immunofluorescence Staining for Visualizing Proliferated Neural Stem Cells in Mouse Brain Sections

Take fixed mouse brain sections containing neural cells, including proliferated neural stem cells labeled with EdU, a thymidine analog.

Outline the sections with a hydrophobic marking to prevent solution spread.

Add a detergent to permeabilize the tissue. Wash to remove excess detergent.

Add a fluorophore to label the EdU. Remove the excess fluorophores.

Using a fluorescence microscope, confirm proper staining of the EdU-labeled cells.

Apply a blocking buffer containing proteins to prevent non-specific antibody binding. Remove the excess proteins.

Incubate with primary antibodies that bind to specific proteins in neural stem cells. Remove the unbound antibodies.

Add fluorophore-conjugated secondary antibodies to bind the primary antibodies. Wash off the unbound antibodies.

Stain the nuclei with a dye and remove the excess dye.

Remove the hydrophobic marking, apply mounting media, and place a coverslip.

Using a confocal microscope, observe the sections to visualize the labeled neural stem cells.