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CAS:596-09-8
Appearance:Light yellow to yellow solid
Storage:Powder : -20℃, 2 years;In solvent(mother liquid): -20℃, 1 month; -80℃, 6 months
Purity:HPLC≥98%
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It is a fluorescent probe used In Vivo staining, and is often used in combination with PI to detect cell viability and cytotoxicity. Ex=494nm / Em=521nm
FDA is a non-fluorescent hydrophobic luciferin derivative, which can penetrate the cell membrane into the cell and produce the fluorescence product luciferin with high intensity through the hydrolysis of diacetate groups catalyzed by cellular lactase. Fluorescent molecules accumulate inside cells with intact cell membranes, so this green fluorescence can serve as a kind of label for cell viability. Cells that do not have a complete cell membrane or metabolic activity cannot accumulate fluorescence products in the cell and therefore cannot display green fluorescence. FDA can be used in combination with the dye PI, inactive dead cells are colored by PI and emit red fluorescence; The active cell PI cannot be colored and, under the action of FDA, tightly fluoresces green. These two colors can well distinguish dead cells from living cells, and provide a more accurate quantitative detection method of living cells than a single color detection.
Examples of using this product（for reference only）
In Vitro:
Cell （KB cells；5μg/mL FDA；15s）:
PI and FDA co-staining experiments evaluated the effect of PTT In Vitro
. KB cells were cultured with Au NCs in 6-well plate at concentration of 100μg/mL based on the content of Au, and then irradiated by 660 nm laser with laser power density of 2 W/cm2 for different times of 0, 5, 10 and 15 min. FDA of 5μg/mL was used to stain the living cells, and the staining time was 15 s. Meanwhile, PI of 5 μg/mL was used to stain the apoptosis cells, and the staining time was 15 min.
Reference：
Chen Q, Guan G, Deng F, Yang D, Wu P, Kang S, Sun R, Wang X, Zhou D, Dai W, Wang X, Zhang H, He B, Chen D, Zhang Q. Anisotropic active ligandations in siRNA-Loaded hybrid nanodiscs lead to distinct carcinostatic outcomes by regulating nano-bio interactions. Biomaterials. 2020 Apr 3;251:120008. doi: 10.1016/j.biomaterials.2020.120008. Epub ahead of print. PMID: 32388031.
Cell（PC12 cell；10 μg/mL FDA；15 min）:
PC12 cells were added to the 6-well plate with a density of 1×105 cells/well. Cells were cultured at 37 ℃ for 24 h with 5% CO2. The preincubated Aβ samples with or without 2.00 mg/mL ulvan were added to the cells and cultured for an additional 48 h. After the medium was gently aspirated with a pipet gun, cells with good adherence were incubated with 10 μg/mL FDA and 5 μg/mL PI for 15 min. Finally, the FDA/PI staining solution was removed, andn the cells were washed three times with PBS buffer and observed using a BX53 fluorescence microscope.
Reference：
Liu F, Zhao W, Zhao F, Dong Q, Wang Y, Wei W, Jia L, Li L, Lu F. Dual Effect of the Acidic Polysaccharose Ulvan on the Inhibition of Amyloid-β Protein Fibrillation and Disintegration of Mature Fibrils. ACS Appl Mater Interfaces. 2020 Sep 16;12(37):41167-41176. doi: 10.1021/acsami.0c14292. Epub 2020 Sep 1. PMID: 32818379.
P. ternata seedling(Pinellia ternata root tip cell; 15 μg/mL FDA; 10min):
We accurately weigh 100 mg of fluorescein diacetate (FDA) (Solarbio, Beijing, China) and dissolved it in 4 mL of acetone to prepare a 25 mg·mL-1 mother liquor, which was stored away from light at -20 ℃. When in use, FDA storage solution was added to 0.65 mol·L-1 mannitol to obtain FDA dyeing solution in proportion. 10 mg of propidium iodide (PI) (Solarbio, China) was dissolved in 2 mL of 0.65 mol·L-1 mannitol to prepare a 5 mol/L mother liquor. The FDA (15 μg·mL-1) and PI (5 μg·mL-1) mixtures were used to treat P. ternata seedlings. P. ternata seedlings were hydroponically cultured and treated with M-Pa-B for 1, 2, 3, and 4 h. After that, the root tips of seedlings were washed three times with deionized water and then dyed with the FDA-PI in the dark for 10 min. The excess dye was washed away thoroughly with deionized water after dyeing. Ten root samples were collected from five individual plants at each time point. A laser confocal scanning microscope was used for observation and photography. The excitation and emission wavelengths are 488 nm and 630 nm, respectively.
Reference：
He Z, Wang Y, Yan Y, Qin S, He H, Mao R, Liang Z. Dynamic analysis of physiological indices and transcriptome profiling revealing the mechanisms of the allelopathic effects of phenolic acids on Pinellia ternata. Front Plant Sci. 2022 Oct 18;13:1039507. doi: 10.3389/fpls.2022.1039507. PMID: 36340387; PMCID: PMC9635339.
