Detecting Cell Death Mechanisms via Hyperthermia

To further determine the type of cell death induced by HT, Capan-2 and BxPC-3 cells were labeled with an apoptosis/necrosis detection kit (ab176749, Abcam, Cambridge, UK). Cells were stained according to the manufacturers’ protocol immediately after removal from the hyperthermic conditions. This kit enables discrimination between healthy, apoptotic, and necrotic cells. Cells incubated at 55 °C for 1 h were used as a positive control for necrosis. Apoptosis was calculated relative to the respective cell lines growing at 37 °C, during the logarithmic growth phase. Images were acquired via Nikon Eclipse TI inverted microscope at a magnification of 4x. Quantitative analysis was performed using the Fiji/ImageJ software [42 (link)]. Microscopic evaluation was performed in sextuplicate.