Introduction of Rhodamine Fluorescent Dye


flourescent dyes

Description: Laser dyes, biological dyes. Is a fluorescent dye, catechols.
Alias: 2-(6-Amino-3-imino-3H-xanthen-9-yl)benzoic acid methyl ester
Appearance: Red to brown powder.
Solubility: soluble in water, soluble in ethanol, reference concentration 1mg/ml.


1. Rhodamine 6G
Alias: rose red 6G, rhodamine 590, yellow light alkaline core fragrant red
Molecular formula: C28H31N2O3Cl
Molecular weight: 479.01

2. Rhodamine 123
English name: 2-(6-Amino-3-imino-3H-xanthen-9-yl)benzoic acid methyl ester
Molecular formula: C21H17ClN2O3
Molecular weight: 380.82

3. Rhodamine B
English name: Rhodamine B
Alias: Rose Red B
Rhodamine B
Molecular formula: C28H31ClN2O3
Molecular weight: 479.01
Rhodamine storage conditions: dry at room temperature

Rhodamine fluorescent dye properties:

1. Appearance: reddish brown powder
2. Purity: ≥95% (HPLC)
3. Product description: Rhodamine 123 is a fluorescent dye that can selectively stain mitochondria of living cells through the cell membrane, showing yellow-green fluorescence. It is widely used to detect mitochondrial membrane potential and is also commonly used to detect apoptosis. It can quickly pass through the cell membrane, can be captured by active mitochondria in just a few minutes, and has no toxicity to the cell.
Rh123 is used to stain many types of cells, including plant cells and bacteria. Since there is a correlation between the amount of intracellular ATP and the fluorescence intensity of Rh123, Rh123 was used to detect intracellular ATP. Rh123 is also used in cancer research.
The maximum excitation wavelength of Rh123 is 507 nm, and the maximum emission wavelength is 529 nm.
4. Dyeing procedure:
(1) Dissolve 0.4mgRh123 into 1mL DMSO to prepare a 1mMRh123-DMSO solution.
(2) Prepare cells with glass slides. The number of cells should be 5×104~5×105 cells/mL.
(3) Incubate the slide and wash the cells with PBS or Hank’s solution.
(4) Dilute 1mM Rh123 solution with culture medium to prepare 1~20µM Rh123 buffer.
(5) Add Rh123 buffer to slides and incubate at 37°C for 30 minutes to 1 hour.
(6) Remove the Rh123 buffer and wash the cells with medium (for fixation after washing the cells, add 10% buffered formalin and incubate for 15-20 minutes, followed by washing with PBS).
(7) Observe the cells with a fluorescence microscope with a fluorescein filter.
Storage conditions: Store at room temperature away from light, dissolve in DMSO to prepare a mother solution and store at -20°C.

Rhodamine fluorescent dye application

(1) Copper ion probe
Copper is an important trace element in the human body. The lack of copper in the body can lead to metabolic disorders and many diseases, such as increased cholesterol, decreased arterial elasticity, and increased blood pressure. For a long time, the study of copper ion bioluminescent probes has been a hot topic. et al designed and synthesized a new rhodamine lactam derivative 5 in 2009, and applied it to the detection of Cu2+ in aqueous solution and living cells. The response of this colorimetric probe to copper ions is instantaneous and reversible, and it will not interfere with the colorimetric and fluorescent signals of copper ions in the presence of high concentrations of other metal ions. Special requirements in biomedical and environmental monitoring are met. At present, this kind of probe has been widely used in the detection of copper ion concentration in environmental systems and in the imaging experiment of copper ion distribution in biological living cells, and its excellent comprehensive performance indicates an excellent application prospect.
(2) Iron ion probe
Iron is an essential trace element in the human body. It mainly exists in the form of complex ions in the human body. It forms hemoglobin and myoglobin with heme and proteins, and plays the role of transporting and storing oxygen in the body. Due to the paramagnetism of Fe3+, general Fe3+ fluorescent probes are all fluorescent quenching type, which is not conducive to the fluorescence imaging and in situ detection of Fe3+ in bovine body. Therefore, the design of fluorescence-enhanced Fe3+ fluorescent probes using the closed-ring-opening transition mechanism of rhodamine molecules has gradually attracted attention.
(3) Mercury ion probe
Mercury is a highly toxic metal. Elemental mercury and mercury ions can enter the environment through various ways. Long-term exposure and ingestion by the human body will cause severe nausea, vomiting, abdominal pain, and kidney function damage, which is extremely harmful. Due to the high attention paid to mercury toxicity, the research on Hg2+ fluorescent probes has been increasing in recent years. Xu et al. designed and synthesized a rhodamine sulfur hydrazide for UV and fluorescence detection of Hg2+ in aqueous phase. The stoichiometric ratio of probe and Hg2+ binding was 2:1. Qian et al. designed and synthesized a highly selective rhodamine-like Hg2+ Fluorescent probes can not only realize the fluorescence detection of Hg2+, but also make preliminary judgment on the existence of Hg2+ by using the color reaction. The probe is reversible. When EDTA is added to the color-developing equilibrium system, the purple-red color of the system becomes colorless. In addition, the fast fluorescence response is another feature of this probe. After the addition of Hg2+, stable and strong fluorescence is generated immediately. Compared with similar probes that require a certain equilibration time, this probe is more suitable for real-time environmental or biological samples. analyze.

Other derivatives

There are many rhodamine derivatives used for imaging purposes, for example Carboxytetramethylrhodamine (TAMRA), tetramethylrhodamine (TMR) and its isothiocyanate derivative (TRITC) and, sulforhodamine 101 (and its sulfonyl chloride form Texas Red) and Rhodamine Red. TRITC is the base rhodamine molecule functionalized with an isothiocyanate group (−N=C=S), replacing a hydrogen atom on the bottom ring of the structure. This derivative is reactive towards amine groups on proteins inside cells. A succinimidyl-ester functional group attached to the rhodamine core, creating NHS-rhodamine, forms another common amine-reactive derivative.

Other derivatives of rhodamine include newer fluorophores such as Alexa 546, Alexa 633, DyLight 550 and DyLight 633, HiLyte fluor 555 HiLyte 594, Janelia Dyes JF549 and JF669 have been tailored for various chemical and biological applications where higher photostability, increased brightness, different spectral characteristics, or different attachment groups are needed.