Rhodamine dyes are membrane-permeable cationic fluorescent probes that specifically recognize mitochondrial membrane potentials, thereby attaching to mitochondria and producing bright fluorescence, and at certain concentrations, rhodamine dyes have low toxicity to cells, so they are commonly used to detect mitochondria in animal cells, plant cells, and microorganisms. Storage: protect from light.

1.Rhodamine 800工作液的配制
1.1 制备储存液
用 525 μL 无水DMSO 稀释 1mg Rhodamine 800 去配制 5 mM 的储备液。
1.2 工作液的配制
用预热好的无血清细胞培养基或 PBS 稀释储存液，配制成 1-20 μM 的 Rhodamine 800工作液。
注：请根据实际情况调整 Rhodamine 800工作液浓度，且现用现配。
2. 细胞染色(悬浮细胞)
2.1 离心收集细胞，加入 PBS 洗涤两次，每次 5 分钟。细胞密度在1×10⁶/mL。
2.2 加入 1 mL Rhodamine 800 工作液，室温孵育 30-60 分钟。
2.3 400 g，离心 3-4 分钟，弃去上清。
2.4 加入 PBS 洗涤细胞两次，每次 5 分钟。
2.5 用 1 mL 无血清培养基或 PBS 重悬细胞后，使用荧光显微镜或流式细胞仪进行观察。
3. 细胞染色(贴壁细胞)
3.1 将贴壁细胞培养于无菌盖玻片上。
3.2 从培养基中移出盖玻片，吸除多余培养基。
3.3 加入 100 μL 染料工作液，轻轻晃动使其完全覆盖细胞，孵育 5-30 分钟。
3.4 吸去染料工作液，用培养基洗 2-3 次，每次 5 分钟 ，使用荧光显微镜或流式细胞仪进行观察。

西域 has not independently confirmed the accuracy of these methods. They are for reference only.

• . Sakanoue J, et al. Rhodamine 800 as a probe of energization of cells and tissues in the near-infrared region: a study with isolated rat liver mitochondria and hepatocytes. J Biochem. 1997 Jan;121(1):29-37.

  . Sakanoue J, et al. Rhodamine 800 as a probe of energization of cells and tissues in the near-infrared region: a study with isolated rat liver mitochondria and hepatocytes. J Biochem. 1997 Jan;121(1):29-37.  [Content Brief]

  . Jin T. et al. Near-infrared fluorescence detection of acetylcholine in aqueous solution using a complex ofrhodamine 800 and p-sulfonatocalix[8]arene. Sensors (Basel). 2010;10(3):2438-49.  [Content Brief]