工作原理

• 即用型

• 快速

• 灵敏

• 适用于3D细胞培养

• 成本低

本试剂盒使用吖啶橙（AO）和碘化丙啶（PI）两种核染色（核酸结合）染料。AO可渗透活细胞和死细胞，可染色所有有核细胞产生绿色荧光；PI只穿透细胞膜受损的有核细胞的细胞膜，并染色死亡细胞产生红色荧光。由于猝灭作用，当细胞同时被AO和PI染色时，所有活的有核细胞产生绿色荧光，所有死的有核细胞产生红色荧光。

工作方法

Data and References

Figure 1. Live-dead cell viability analysis by using Cyto3D Live-Dead Assay Kit.
Glioblastoma cells (SF 298, about 60% cell viability) were 3D cultured in VitroGel system for 2 days. 2 μL of Cyto3D reagent was added to each well containing 50 μL hydrogel and 50 μL cover medium.  The mixture was incubated at 37 °C for 5-10 min. The cells were then observed under a fluorescent microscope. The images show the Live (green) and Dead (orange) cells within the 3D hydrogel matrix. The z-stack images of cells within hydrogel were then 3D reconstructed and showed in the 4D view images. The live and dead cells at higher levels of the hydrogel clearly show in the images by using Cyto3D Live-Dead Assay Kit.

Figure 2. Live-dead cell viability images of stem cell spheroids.
Stem cells were static suspension-cultured in VitroGel STEM (CAT# VHM02) for 5 days. 2 μL of Cyto3D reagent was added to each well containing 100 μL cell suspension.  The mixture was incubated at 37 °C for 5-10 min. The cells were then observed under a fluorescent microscope. The images show the Live (green) and Dead (orange) stem cell spheroids cultured in a 3D hydrogel matrix. The live-dead dyes of Cyto3D Live-Dead Assay Kit can successfully penetrate into large cell spheroids for cell viability analysis.