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  3. DiI

DiI is a long-chain carbocyanine dye. Carbocyanine dyes are widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins.

For research use only. We do not sell to patients.

DiI Chemical Structure

DiI Chemical Structure

CAS No. : 41085-99-8

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Solid + Solvent
10 mM * 1 mL in DMSO
ready for reconstitution
USD 34 In-stock
Solution
10 mM * 1 mL in DMSO USD 34 In-stock
Solid
5 mg USD 25 In-stock
10 mg USD 30 In-stock
25 mg USD 63 In-stock
50 mg USD 110 In-stock
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Customer Review

Based on 15 publication(s) in Google Scholar

Top Publications Citing Use of Products

    DiI purchased from MedChemExpress. Usage Cited in: J Immunother Cancer. 2020 Aug;8(2):e000330.  [Abstract]

    Ecto-CRT translocation to cell surfaces confirmed by three counterstains and merged analysis with an anti-CRT antibody followed by FITG-conjugated secondary antibody (green), DiI (red), and 4',6-diamidino-2-phenylindole (DAPI, blue).
    • Biological Activity

    • Purity & Documentation

    • References

    • Customer Review

    Description

    DiI is a long-chain carbocyanine dye. Carbocyanine dyes are widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins[2].

    In Vitro

    Carbocyanine dyes are widely used as Di to label cells, organelles, liposomes, viruses and lipoproteins. Long-chain carbocyanines which include DiO (DiOC18(3)), DiI (DiIC18(3)), DiD (DiIC18(5)) and DiR, and dialkyl aminostyryl dye DiA (4-Di-16-ASP) are used for labeling membranes and other hydrophobic structures. DiIC16(3) has shorter alkyl substituents (C16) than DiI (C18). They have extremely high extinction coefficients, environmental dependent fluorescence and short excited-state lifetimes in lipid environments. They are oils at room temperature and weakly fluorescent in water but highly fluorescent and quite photostable when incorporated into membranes or bound to lipophilic biomolecules. These optical characteristics make them ideal for staining the cytoplasmic membranes of cells. Once applied to cells, these dyes diffuse laterally within the plasma membrane, resulting in staining of the entire cell[1].
    DiO, DiI, DiD and DiR exhibit distinct green, orange, red and infrared fluorescence, respectively? thus facilitating multicolor imaging and flow cytometric analysis of live cells . DiO and DiI can be used with standard FITC and TRITC filters respectively. Among them DiI and its analogs are most frequently used since they usually exhibit very low cell toxicity. In addition, DiI is widely used for determining lipoproteins such as LDL and HDL. The lipophilic aminostyryl dye DiA is also often used for neuronal tracing[2].
    General Protocol
    1. Preparing Stain Solutions of Di
    a. Prepare DMF, DMSO or ethanol stock solutions: The stock solutions should be prepared in dimethyl formamide (DMF), dimethylsulfoxide (DMSO, or ethanol DMSO at 1-5 mM. DMF is preferable to ethanol as a solvent for Di. The stock solution should be used promptly. Any unused solution need to be aliquoted and refrozen at least -20℃. Avoid repeated freeze/thaw cycle. The solution can be stored for 6 months.
    b. Prepare working solutions: Dilute the stock solutions into a suitable buffer such as serum-free culture medium, HBSS or PBS to make 1 to 5 μM working solutions. We do not recommend storing the aqueous solution for more than one day.
    Note: The final concentration of the working solution should be empirically determined for different cell types and/or experimental conditions.
    2. Suspension cells
    a. Centrifuge at 1000 g at 4℃ for 3-5 minutes and then discard the supernatant. Wash twice with PBS, 5 minutes each time.The cell density is 1×106/mL.
    b. Add 1 mL of Di working solution, and then incubate at room temperature for 5-30 minutes.
    c. Centrifuge at 400 g at 4℃ for 3-4 minutes and then discard the supernatant.
    d. Wash twice with PBS, 5 minutes each time.
    e. Resuspend cells with serum-free cell culture medium or PBS.Observation by fluorescence microscopy or flow cytometry.
    3. Adherent cells
    a. Culture adherent cells on sterile coverslips.
    b. Remove the coverslip from the medium and aspirate excess medium.
    c. Add 100 μL of working solution, gently shake it to completely cover the cells, and then incubate at room temperature for 5-30 minutes.
    d. Wash twice with medium, 5 minutes each time. Observation by fluorescence microscopy or flow cytometry.

    MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

    In Vivo

    DiI-labeled motoneurons have remained viable for up to 4 weeks in culture and up to one year in vivo[1]

    MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

    Molecular Weight

    933.87

    Formula

    C59H97ClN2O4

    CAS No.
    Appearance

    Solid

    Emission (Em)

    565

    Excitation (Ex)

    549

    SMILES

    CCCCCCCCCCCCCCCCCC[N+]1=C(/C=C/C=C2N(CCCCCCCCCCCCCCCCCC)C3=C(C=CC=C3)C/2(C)C)C(C)(C)C4=C1C=CC=C4.O=Cl(=O)([O-])=O

    Shipping

    Room temperature in continental US; may vary elsewhere.

    Storage

    4°C, sealed storage, away from moisture and light

    *In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

    Solvent & Solubility
    In Vitro: 

    DMSO : 12.5 mg/mL (13.39 mM; ultrasonic and warming and heat to 80°C; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)

    Preparing
    Stock Solutions
    Concentration Solvent Mass 1 mg 5 mg 10 mg
    1 mM 1.0708 mL 5.3541 mL 10.7081 mL
    5 mM 0.2142 mL 1.0708 mL 2.1416 mL
    View the Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

    • Molarity Calculator

    • Dilution Calculator

    Mass (g) = Concentration (mol/L) × Volume (L) × Molecular Weight (g/mol)

    Mass
    =
    Concentration
    ×
    Volume
    ×
    Molecular Weight *

    Concentration (start) × Volume (start) = Concentration (final) × Volume (final)

    This equation is commonly abbreviated as: C1V1 = C2V2

    Concentration (start)

    C1

    ×
    Volume (start)

    V1

    =
    Concentration (final)

    C2

    ×
    Volume (final)

    V2

    In Vivo:

    Select the appropriate dissolution method based on your experimental animal and administration route.

    For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
    To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day.
    The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.

    • Protocol 1

      Add each solvent one by one:  10% DMSO    40% PEG300    5% Tween-80    45% Saline

      Solubility: 1.25 mg/mL (1.34 mM); Suspended solution; Need ultrasonic

      This protocol yields a suspended solution of 1.25 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (12.5 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.

      Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
    • Protocol 2

      Add each solvent one by one:  10% DMSO    90% (20% SBE-β-CD in Saline)

      Solubility: 1.25 mg/mL (1.34 mM); Suspended solution; Need ultrasonic

      This protocol yields a suspended solution of 1.25 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.

      Taking 1 mL working solution as an example, add 100 μL DMSO stock solution (12.5 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.

      Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
    In Vivo Dissolution Calculator
    Please enter the basic information of animal experiments:

    Dosage

    mg/kg

    Animal weight
    (per animal)

    g

    Dosing volume
    (per animal)

    μL

    Number of animals

    Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
    Please enter your animal formula composition:
    %
    DMSO +
    +
    %
    Tween-80 +
    %
    Saline
    Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
    The co-solvents required include: DMSO, . All of co-solvents are available by MedChemExpress (MCE). , Tween 80. All of co-solvents are available by MedChemExpress (MCE).
    Calculation results:
    Working solution concentration: mg/mL
    Method for preparing stock solution: mg drug dissolved in μL  DMSO (Stock solution concentration: mg/mL).

    *In solvent : -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light)

    The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
    Method for preparing in vivo working solution for animal experiments: Take μL DMSO stock solution, add μL . μL , mix evenly, next add μL Tween 80, mix evenly, then add μL Saline.
     If the continuous dosing period exceeds half a month, please choose this protocol carefully.
    Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
    Purity & Documentation

    Purity: 99.92%

    Dyeing Example
    References

    Complete Stock Solution Preparation Table

    * Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
    Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (sealed storage, away from moisture and light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

    Optional Solvent Concentration Solvent Mass 1 mg 5 mg 10 mg 25 mg
    DMSO 1 mM 1.0708 mL 5.3541 mL 10.7081 mL 26.7703 mL
    5 mM 0.2142 mL 1.0708 mL 2.1416 mL 5.3541 mL
    10 mM 0.1071 mL 0.5354 mL 1.0708 mL 2.6770 mL
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    DiI Related Classifications

    Help & FAQs
    • Do most proteins show cross-species activity?

      Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.

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