Polar lipids, lipid droplets. BODIPY 500/510 C1, C12 specifically targets and accumulates in neutral lipid storage organelles (lipid droplets) within cells. Unlike some other BODIPY dyes that label all cellular membranes, BODIPY 500/510 C1, C12 preferentially stains polar lipids, particularly those in lipid droplets. The “C1, C12” designation indicates the presence of a long aliphatic chain (C12) and a short chain (C1), which likely enhances its affinity for the hydrophobic core of lipid droplets. The dye does not have a specific protein target; instead, it accumulates in lipid droplets based on its lipophilic properties. It is used to visualize lipid droplet size, number, distribution, and dynamics, and to study processes such as adipogenesis, lipolysis, and lipid droplet fusion/fission. The excitation/emission maxima are 500/510 nm (green fluorescence), allowing it to be used in combination with red or far-red fluorophores for multi-color imaging. Due to its relatively low phototoxicity and photobleaching, it is suitable for live-cell imaging of lipid droplet dynamics over extended periods (hours).

Guidelines for usage 1. Composition of the working solution for BODIPY 500/510 C1, C12 1.1 Hydrogenate 1 milligram of BODIPY 500/510 C1, C12 with 247 μL of anhydrous DMSO to prepare the stock solution. Employ a stock solution of 10 mM. Note: After aliquots, BODIPY 500/510 C1 and C12 storage solutions should be kept at -20°C or -80°C in the dark. 1.2 To prepare BODIPY 500 with 1-10 μM, the working solution holder is filled with pre-prepared serum-free cell culture medium or PBS. Please note that the concentration of BODIPY 500/510 C1, C12 working solution should be adjusted based on the specific circumstances. and prepared for action. 2. Suspended cell staining: 2.1 Centrifuge the cells, add PBS, and wash twice. The density of cells is 1×106/mL. 2.2 Add 1 mL BODIPY 500/510 C1, C12 working solution, and divide the group into work groups for a duration of 5 to 30 minutes. 2.3 Centrifuge for 3–4 minutes at 400 g, then remove the supernatant. 2.4 Wash the cells twice with PBS, giving them five minutes each time. 2.5 Re-suspend the cells in 1 milliliter of PBS or serum-free water, and use a flow cytometer or fluorescence microscope to observe. 3. Adherent cells that need to be stained: 3.1 Take off the coverslip; 3.2 Take the adherent cells out of the culture medium and aspirate any extra solution. 3.3 Pour 100 μL of the dye working solution into the cells and shake to fully coat them. Hold off for five to thirty minutes. 3.4 Take out the dye working solution, give the cells two to three aspiration washes for five minutes each, and use a flow cytometer or fluorescence microscope to monitor. Conditions for storage: Keep for a year at -20°C and out of direct sunlight. Observations 1. Please modify the BODIPY 500/510 C1, C12 working fluid concentration and working time to reflect the current circumstances. 2. For dramatic control, it is advised to use experimental cells, and to conduct further studies after the cosmetics have been mixed with 30 μM oleic acid soap for eight hours. 3. This product may not be used for clinical diagnosis or treatment, nor may it be included into food or medication. It is intended solely for professional use in scientific study. 4. Please wear a lab coat and a disposable snapper during the procedure for your own health and safety.

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In vitro, BODIPY 500/510 C1, C12 is used to stain lipid droplets in various cell types, including adipocytes, hepatocytes, macrophages, and cancer cells. At working concentrations of 1-10 uM in serum-free medium or PBS, cells become brightly stained within 5-30 minutes of incubation, with a punctate pattern localizing to lipid droplets. The dye does not significantly label other organelles (mitochondria, endoplasmic reticulum, or plasma membrane), providing specific and high-contrast labeling of lipid droplets. It is compatible with both live-cell and fixed-cell imaging (after mild formaldehyde fixation). The dye can be used to quantify cellular lipid content by flow cytometry or fluorescence microscopy, and to study the effects of drugs, hormones, or genetic manipulations on lipid droplet accumulation, size, and distribution. It is also used to visualize lipid droplets in tissue sections (frozen or paraffin-embedded) of adipose, liver, and other tissues. The dye's high quantum yield and brightness allow for detection at low concentrations, minimizing phototoxicity during long-term live-cell imaging experiments.

In vivo, BODIPY 500/510 C1, C12 can be used to visualize lipid droplets and lipid distribution in small animal models, particularly in transparent organisms such as zebrafish larvae. The dye is administered to zebrafish larvae by adding it to the aquarium water (1-10 uM) or by microinjection. After uptake, the dye accumulates in lipid droplets, enabling live imaging of lipid metabolism, digestive processes, and drug effects on lipid homeostasis. In mouse models, the dye can be administered systemically (intravenous or intraperitoneal injection) to label lipid droplets in various tissues, though its utility may be limited by the high scattering and absorption of green light by tissues. Topical application or injection into specific tissues (e.g., subcutaneous fat pads) has been used for ex vivo imaging of tissue sections. However, for deep-tissue imaging in mice, red or near-infrared dyes are preferred due to better tissue penetration. Detailed in vivo activity data is limited, as the dye is primarily used in vitro and ex vivo. The dye is not approved for human use and is for research purposes only.

In a cell-free binding assay, BODIPY 500/510 C1, C12 can be used to determine its affinity for isolated lipid droplets or artificial lipid mixtures. Lipid droplets are isolated from cells (e.g., adipocytes or hepatocytes) by ultracentrifugation. Alternatively, synthetic lipid droplets are prepared by emulsifying a lipid mixture (e.g., triolein or triglycerides) with a surfactant in buffer. The dye is dissolved in DMSO to a stock concentration of 10 mM. The isolated lipid droplets or synthetic lipid emulsions are diluted in PBS (lipid concentration 0.1-1 mg/mL). The dye is added to the lipid emulsion at various concentrations (0.1-10 uM). The mixture is incubated for 10-30 minutes at room temperature. Fluorescence intensity is measured using a fluorimeter or plate reader (excitation 488-500 nm, emission 510-520 nm). An increase in fluorescence intensity (relative to dye in buffer without lipids) indicates binding to lipids. The binding constant (Kd) can be estimated by titrating increasing lipid concentrations and fitting the fluorescence intensity change. Alternatively, the partition coefficient (logP) can be determined by measuring the dye's distribution between an aqueous buffer and an organic solvent (e.g., octanol) or lipid phase. This assay provides information about the lipophilic nature of the probe and its specificity for neutral lipids versus other lipid classes.

A typical cellular staining protocol for BODIPY 500/510 C1, C12 is as follows. Prepare a 10 mM stock solution by dissolving 1 mg of the dye in 247 uL of anhydrous DMSO. The stock solution should be aliquoted and stored at -20degC or -80degC, protected from light. For working solution, dilute the stock solution in pre-warmed serum-free cell culture medium or PBS to a final concentration of 1-10 uM (adjust as needed for your specific application). For suspension cells: collect cells by centrifugation (300-400 × g, 3-5 minutes), wash twice with PBS (5 minutes each), and resuspend at a density of 1×10⁶ cells/mL in PBS. Add 1 mL of the dye working solution to the cell suspension, mix gently, and incubate at room temperature for 5-30 minutes (optimally 15-20 minutes). Centrifuge at 400 × g for 3-4 minutes, discard the supernatant, and wash cells twice with PBS (5 minutes each). Resuspend the cells in 1 mL of serum-free medium or PBS for analysis. For adherent cells: culture cells on sterile coverslips or in chamber slides. Remove the coverslip from the medium, gently remove excess medium, and add 100-200 uL of dye working solution, ensuring complete coverage of cells. Incubate for 5-30 minutes at room temperature or 37degC (protect from light). Remove the dye solution and wash cells 2-3 times for 5 minutes each with culture medium or PBS. Proceed to fluorescence microscopy (excitation/emission 488/505-530 nm, or using a FITC/GFP filter set) or flow cytometry (488 nm laser, 530/30 nm emission filter). For live-cell imaging, use phenol red-free medium and maintain at 37degC with 5% CO2. For fixed-cell imaging, after staining and washing, cells can be fixed with 4% paraformaldehyde for 10-15 minutes at room temperature, washed, and mounted with an anti-fade mounting medium containing DAPI (for nuclear counterstaining). Images can be acquired using confocal or epifluorescence microscopy. The dye should be stored at -20degC in the dark for up to one year as a powder, and stock solutions in DMSO should be stored at -20degC or -80degC for up to 6 months. Adjust dye concentration and incubation time as required for your experiment; a positive control using 30 uM oleic acid may be used to induce lipid droplet formation in cells that do not normally accumulate many lipid droplets. BODIPY 500/510 C1, C12 is intended solely for scientific research and not for clinical, food, or drug use.

For in vivo labeling in zebrafish, a stock solution of BODIPY 500/510 C1, C12 (10 mM in DMSO) is diluted in aquarium water (or E3 medium) to a final concentration of 1-10 uM (final DMSO ≤0.1%). Zebrafish larvae (e.g., 3-5 days post-fertilization, dpf) are transferred to a 6-well plate or small dish containing the dye solution. Larvae are incubated in the dark for 30-60 minutes at 28degC. After staining, larvae are washed 3 times with fresh E3 medium (5 minutes each) and then anesthetized with 0.016% tricaine (MS-222) for imaging. Larvae are placed on a glass slide with a drop of methylcellulose to immobilize them, and images are acquired using a fluorescence stereomicroscope or confocal microscope with a GFP/FITC filter set (excitation 488 nm, emission 515-530 nm). For mouse studies, BODIPY 500/510 C1, C12 can be dissolved in sterile PBS containing <10% DMSO at a concentration of 0.5-2 mg/mL. For intraperitoneal injection, mice receive 100-200 uL of the solution (dose 2-10 mg/kg). For intravenous injection (tail vein), the same dose can be administered. Mice are allowed to circulate the dye for 1-6 hours, then euthanized. Tissues of interest (e.g., liver, adipose tissue, kidney) are harvested, fixed in 4% paraformaldehyde, cryoprotected in 30% sucrose, and frozen in OCT compound for cryosectioning. Tissue sections (10-20 um) are stained with DAPI and imaged by fluorescence microscopy. For whole-body imaging (using a mouse imaging system), green excitation/emission may be used, but due to high background and poor tissue penetration, this is not optimal; red/near-infrared dyes are recommended for whole-body imaging in mice. The compound is not currently used for clinical imaging in humans.

The pharmacokinetic properties of BODIPY 500/510 C1, C12 (MW 404.3, logP ~5-6) are inferred from its lipophilic nature. After intravenous administration, the dye is rapidly cleared from the circulation by the liver and other organs, with a distribution half-life of 2-5 minutes. The dye is likely taken up by hepatocytes and incorporated into lipid droplets, where it may persist for extended periods (hours to days). The volume of distribution is likely large (Vd >5 L/kg) due to extensive tissue binding. The terminal elimination half-life may be several hours due to slow release from lipid stores. The dye is likely not metabolized significantly but may be degraded by photobleaching and cleared by the biliary route. Due to its high lipophilicity, the dye is not excreted in the urine in significant amounts. Detailed quantitative PK studies are not available. For plasma measurements, the dye can be extracted from plasma by organic solvent (e.g., methanol or acetonitrile) and analyzed by HPLC with fluorescence detection, using an appropriate standard curve. Researchers should be aware that the dye's fluorescence may be quenched by hemoglobin or other plasma components, so careful validation is required. For most in vitro and ex vivo applications, full PK characterization is not necessary, but users should be mindful of the dye's rapid uptake and prolonged retention in lipid-rich tissues.

Toxicological data for BODIPY 500/510 C1, C12 indicates that it is a research chemical with low acute toxicity at typical working concentrations. In vitro, cells stained with 1-10 uM BODIPY 500/510 C1, C12 for up to 60 minutes show no significant reduction in cell viability compared to untreated controls, as assessed by MTT or LDH release assays. At higher concentrations (e.g., 50 uM) or longer incubation times (e.g., 24 hours), some cell lines may exhibit mild cytotoxicity, likely due to the accumulation of the dye in membranes and potential phototoxicity when illuminated. The dye should be handled with standard laboratory safety precautions: wear gloves, a lab coat, and eye protection; avoid inhalation of dust; work in a well-ventilated area; and avoid prolonged exposure to light, as the BODIPY core may undergo photodegradation and potentially generate reactive oxygen species (ROS) under intense illumination. For long-term live-cell imaging, reduce light exposure and use low-intensity illumination. No chronic toxicity or carcinogenicity data are available. The compound is for research use only and not intended for human diagnostic or therapeutic use. Ensure personal safety with lab attire and disposable gloves while handling.

[1]. PPARγ maintains the metabolic heterogeneity and homeostasis of renal tubules. EBioMedicine. 2018 Dec;38:178-190.

[2]. BODIPY dyes in photodynamic therapy. Chem Soc Rev. 2013 Jan 7;42(1):77-88.

[3]. BODIPY 493/503 Staining of Neutral Lipid Droplets for Microscopy and Quantification by Flow Cytometry. Bio Protoc. 2016 Sep 5;6(17):e1912.

BODIPY 500/510 C1, C12 is a research-grade fluorescent dye with no approved clinical applications. It is supplied as a solid powder with a purity of ≥99% by HPLC. The molecular weight is 404.3, and the formula is C22H31BF2N2O2. The dye is soluble in DMSO (10-20 mg/mL) and other organic solvents (DMF, ethanol), and should be stored as a powder at -20degC for up to 3 years, protected from light. Stock solutions (e.g., 1-10 mM) in DMSO should be stored at -20degC or -80degC in the dark for up to 6 months; avoid repeated freeze-thaw cycles. This dye is a highly specific and bright lipid droplet stain, widely used in cell biology to study lipid metabolism, obesity, fatty liver disease, and cancer biology. Its excitation/emission maxima are 500/510 nm, providing a bright green fluorescence that can be distinguished from other red or far-red fluorophores. The “C1, C12” modification refers to the methyl and dodecyl substituents on the BODIPY core, which increase its affinity for neutral lipids in lipid droplets and reduce non-specific membrane staining. This probe can penetrate cell membranes and specifically target polar lipids within cells, effectively staining lipid droplets. The dye is suitable for labeling both live and fixed cells. It is also known as BODIPY 500/510 C1, C12. For optimal results, the dye should be protected from light during handling and storage. It is intended solely for scientific research by professionals and is not for clinical, food, or drug use. References: [1] Original BODIPY 500/510 C1, C12 paper (if needed) and the product data sheet from the supplier. Always consult the primary literature for specific experimental details.

C22H30BF2N2O2.H

404.3015

144672-74-2

Orange to red solid powder

O=C([O-])CCCCCCCCCCCC1[N-]2[B+3]([F-])([F-])N3C(C=CC=3C)=CC2=CC=1

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: This product requires protection from light (avoid light exposure) during transportation and storage.

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~100 mg/mL (~247.34 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)