Live-Dead Cell Staining Kit (BioVision)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Incubate for 15 min at 37°C	Observe cells immediately under a fluorescence microscope using a band-pass filter (detects fluorescein and rhodamine).

Protocol tips
Incubate for 15 min at 37°C
Downstream tips
Observe cells immediately under a fluorescence microscope using a band-pass filter (detects fluorescein and rhodamine).

Publication protocol

The damage to tumor cells induced by thermal ablation with SWCNTs-(KFKA)7 and NIR irradiation was evaluated in vitro. Colon26 cells and HepG2 cells (2 × 105 cells/500 µl) were seeded in an 8-well chambered coverglass (Asahi Glass, Tokyo, Japan) and incubated overnight. After changing the culture medium, 1.5 µl and 5 µl of SWCNTs solution (200 µg SWCNTs/ml) was added to the 400 µl of culture medium of colon26 cells and HepG2 with final concentrations of 0.75 µg/ml and 2.5 µg/ml, respectively. After 2 hours of incubation, the wells were exposed to irradiation with an 808 nm NIR laser for 3 min at 1.2 W, collected in a 1.5 ml tube, and stained with a Live-Dead cell staining kit (Biovision, Mountain View, CA). The fluorescence microscopic observation was performed using a Biozero Bz-8000 (Keyence, Osaka, Japan) with Ex/Em = 470/535 nm (Live-Dye fluorescing green) and Ex/Em = 540/605 nm (propidium iodide fluorescing red), respectively. Confocal microscopy was carried out with A1RMP (Nikon, Tokyo, Japan) with Ex/Em = 488/525 nm (green) and Ex/Em = 562/595 nm (red), respectively. For flowcytometric analysis, cells were stained with propidium iodide of a Live-Dead cell staining kit and the number of labeled cells was analyzed by a FACSCant II (BD biosciences, San Jose, CA) with Ex/Em = 488/585 nm.

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