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, andmicroorganisms^[1].

1.TMRM 工作液的配制
1.1 制备储存液
用 525 μL 无水 DMSO 稀释 1mg TMRM 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 TMRM 工作液。
注：请根据实际情况调整 TMRM 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 TMRM 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。

401.48

C₂₅H₂₅N₂O₃

115532-49-5

四甲基罗丹明甲酯

Room temperature in continental US; may vary elsewhere.

Please store the product under the recommended conditions in the Certificate of Analysis.

• 
  Description: TMRM is a rapidly and selectively mitochondrial membrane potential fluorescent probe with red fluorescence.
  Method: For mitochondrial membrane potential measurement.
  1. Cultures are exposed to Millipore-filtered solutions (0.22 μM) containing TMRM and/or drugs for 1 h at 37℃ (except the experiment involving different durations of exposure to TMRM).
  2. After treatment, solutions are removed and growth media reapplied under sterile conditions, and cultures are post-incubated for 18 hours at 37℃.
  3. Use a confocal laser scanning microscope for image.
• 
  Description: TMRM is a rapidly and selectively mitochondrial membrane potential fluorescent probe with red fluorescence.
  Method: For mitochondrial membrane potential measurement.
  1. Testing cells are pretreated first.
  2. Incubate cells with TMRM (20 μM; 30 min; 37℃; dark).
  3. Subsequently, the cells are rinsed with PBS for twice and fixed in 10% formaldehyde neutral buffer solution at room temperature for 10 min.
  4. Cells are imaged under a Zeiss LSM710 Confocal laser-scanning microscope.
• 
  Description: TMRM is a rapidly and selectively mitochondrial membrane potential fluorescent probe with red fluorescence.
  Method: For mitochondrial membrane potential measurement.
  1. Testing cells are pretreated first.
  2. The TMRM fluorescent dye is formulated into 50 nM/L with PBS. Incubate cells with TMRM (50 μL; 25 min; 37℃; dark).
  3. The fluorescence intensity of membrane potential measurement is detected by High Content Cell Imaging Analysis System (IN Cell analyzer 2500HS, GE, USA) and use a confocal laser scanning microscope for image.