Processing of Lentivirus-Transduced Mouse Intestinal Organoids for Immunohistochemical Analysis

Take a multi-well plate containing mouse intestinal organoids embedded in a gel matrix.
The organoid cells are transduced with a reporter gene using lentivirus. This gene expresses a reporter protein.
Add fixative to the well to preserve organoid tissue morphology.
Wash with buffer to remove residual fixative.
Resuspend the organoids in buffer, transfer them to a glass vial, and allow them to settle.
Replace the buffer with ethanol containing eosin dye, which stains the organoids to enable visualization.
Treat the organoids with increasing concentrations of ethanol to dehydrate them.
Add xylene to increase tissue transparency.
Decant the xylene, add molten paraffin, and incubate at high temperature to infiltrate the tissue.
Replace with fresh paraffin, ensuring complete infiltration.
Transfer organoids into a mold containing paraffin, center them, and allow the paraffin to solidify.
Add additional paraffin and insert an embedding cassette.
Paraffin-embedded lentivirus-transduced organoids are ready for immunohistochemical analysis.

Remove the medium from a single well with full-grown organoids, leaving the embedded organoids intact, and add one milliliter of 4% paraformaldehyde in PBS directly