Chemical Name: Rhodamine 123, 2-(6-Amino-3-imino-3H-xanthen-9-yl)benzoic acid methyl ester, hydrochloride [cas# : 62669-70-9]
Appearance: Red to reddish-brown powder or solid
MW: 380.82, C21H17ClN2O3
Storage Condition : 0-5ºC, protect from light
Shipping Condition : ambient temperature
Rhodamine 123 (Rh123) is cell-membrane permeable and localizes in mitochondria of viable cells to emit yellowish-green fluorescence (Fig. 1). Rh123 is utilized for staining a wide variety of cells, including plant cells and bacteria. Since there is a correlation between the amount of ATP in a cell and the fluorescence intensity of Rh123, this compound is used for the detection of intracellular ATP. Rh123 is also used in cancer research.
Fig. 1 Cell staining mechanism
- Dissolve 0.4 mg Rh123 in 1 ml DMSO to prepare 1 mM Rh123-DMSO solution.
- Prepare cells with a glass slide. The cell number will be 5x104 to 5x105 cells per ml.
- Incubate the slide and wash cells with PBS or Hank’s medium.
- Dilute the 1 mM Rh123 solution with culture medium to prepare 1-20 μM Rh123 buffer solution.
- Add the Rh123 buffer solutiona) to the glass slide and incubate at 37oC for 30 min to 1 hour.
- Remove the Rh123 buffer solution and wash cells with culture medium.b)
- Observe the cells under a fluorescence microscope with a fluorescein filter.
a) Incubate the Rh123 buffer solution at 37oC prior to adding to cells.
b) For fixing after washing cells, add 10% formarin buffer and incubate for 15-20 min, and then wash with PBS.
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