Introduction of 5(6)-Carboxyfluorescein Diacetate
5(6)-Carboxyfluorescein diacetate, also known as CFDA, is a fluorescent compound commonly used in cell biology and biochemistry research. It is a cell-permeant substrate that can be cleaved by intracellular esterases to release the fluorescent 5(6)-carboxyfluorescein dye. This dye can then be used to stain and track living cells in various experimental applications, including viability assays, cell tracking, and monitoring cellular uptake and trafficking.
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Design Principle and Characteristics of CFDA
1. Design Principle
The molecular structure of 5(6)-carboxyfluorescein diacetate includes an acetate group, which allows the molecule to be membrane-permeable and able to enter living cells.
Once inside the cell, the acetate groups are cleaved by intracellular esterases, releasing the 5(6)-carboxyfluorescein (5(6)-CF), which is a highly fluorescent compound that can be easily detected and visualized.
2. Characteristics
- Cell-permeable: 5(6)-carboxyfluorescein diacetate can easily penetrate the cell membrane and enter living cells, where it can be hydrolyzed to release the fluorescent dye.
- Selective Labeling: The hydrolysis of 5(6)-carboxyfluorescein diacetate occurs mainly within the cells, allowing for selective labeling of live cells.
- Stability: 5(6)-carboxyfluorescein diacetate is stable in cell culture conditions, which makes it suitable for long-term cell tracking experiments.
- Fluorescent Signal: Upon hydrolysis, the released 5(6)-carboxyfluorescein shows bright and stable fluorescence, making it an ideal dye for imaging studies.
- Compatibility: 5(6)-carboxyfluorescein diacetate is compatible with a variety of imaging techniques, including fluorescence microscopy and flow cytometry.
Overall, the design principle and characteristics of 5(6)-carboxyfluorescein diacetate make it a valuable tool for cell labeling, tracking, and imaging applications in biological research.
Applications of CFDA
CFDA is a widely used fluorescent marker that has numerous applications in various fields, including biology and other industries. Some of its key applications include:
1. Biological Field
- Cell Viability Assays: CFDA is commonly employed in cell viability assays to assess living cells by measuring esterase activity. It is converted into 5(6)-carboxyfluorescein, a highly fluorescent compound, by intracellular esterases in viable cells.
- Cell Labeling and Tracing: CFDA can be used for labeling and tracking cells in biological samples, enabling researchers to study cell migration, proliferation, and differentiation in various biological processes.
- Membrane Permeability Studies: CFDA is used to evaluate cell membrane integrity and permeability in different cell types, making it an essential tool for investigating cellular uptake mechanisms and drug delivery studies.
- Live Cell Imaging: CFDA is utilized for live-cell imaging to visualize dynamic processes within cells in real-time, allowing researchers to monitor cellular behavior, morphology, and interactions under physiological conditions.
2. Other Applications
- Environmental Monitoring: CFDA is used in environmental applications to detect and track pollutants, contaminants, and microbial activity in water bodies, soil, and air samples.
- Food Quality Assessment: CFDA can be employed in the food industry to evaluate food quality, freshness, and safety by monitoring microbial contamination, spoilage, and preservation methods.
- Pharmaceutical Analysis: CFDA is utilized in the pharmaceutical industry for drug screening assays, pharmacokinetic studies, and drug delivery system development, enabling the assessment of drug efficacy, bioavailability, and cellular distribution.
- Chemical Analysis: CFDA is utilized in chemical analysis techniques for fluorescence detection, quantification, and identification of analytes in complex mixtures, contributing to analytical chemistry applications.
Overall, the versatile properties of 5(6)-carboxyfluorescein diacetate make it a valuable tool in various fields, offering numerous applications for biological research, environmental monitoring, food analysis, pharmaceutical studies, and chemical analysis. Its fluorescence properties, cell-permeability, and ease of use make it a preferred choice for diverse applications requiring fluorescent labeling, imaging, and detection.
