Characterization of Small Bowel Neuroendocrine Tumor Spheroids Using Immunofluorescence

Take small bowel neuroendocrine tumor spheroids in a tube.

Add a fixative solution to preserve the cells. 

To wash, centrifuge, and replace the fixative with buffer.

Centrifuge and discard the supernatant to remove any remaining fixative.

Add a buffer containing detergent and a blocking agent to permeabilize the cellular membranes and prevent non-specific antibody binding.

Wash with buffer to remove excess detergent and blocking agent.

Incubate with primary antibodies, which bind to the target protein specific to the neuroendocrine cell.

Wash with buffer to remove unbound antibodies.

Add fluorophore-tagged secondary antibodies that bind to the primary antibodies.

Rinse with buffer to remove excess antibodies.

Add a mounting medium containing a dye to stain the nucleus.

Transfer the spheroids onto a slide and mount with a cover slip.

Use a Fluorescence microscope to visualize neuroendocrine cells of the spheroid expressing specific proteins.