Live Imaging for Quantifying Radiation Therapy Effects on Patient-Derived Tumor Organoids

To begin, obtain the patient-derived tumor organoid, or PDTO, Matrigel Dome and position it in the live imaging system. In the IncuCyte software, select Schedule To Acquire. Click on the plus sign in the upper left corner, select Scan On Schedule, and click Next.

Then select Create Vessel New, choose Organoid, followed by QC and Phase plus Brightfield image channels. After choosing the software validated plate, as per the catalog number, select the location of the plate on the instrument and the wells according to the plate layout. To create plate map, click on the plus sign in the upper right corner of the plate layout.

Then under the Cells section, click the Add or Edit Cell Type icon and indicate the cell type. Choose the appropriate wells and click on the plus icon. To indicate the treatment, go to growth conditions and click on Create a New Condition.

Apply to the appropriate wells with the plus icon. Click on OK, followed by Next. After checking if defer analysis until later is selected, click on Next.

Choose to scan twice a day indefinitely. Finally, check the summary and click on Add To Schedule. Irradiation of one of the breast cancer PTDOs in a single dose of two Gray radiation therapy significantly delayed PDTO regrowth, while higher radiation doses completely prevented the regeneration of growing PDTO.