Fura-4F AM

Operating Instructions
(The following is a recommended experimental protocol for guidance only, and adjustments are required based on your specific needs)

1. Stock Solution Preparation
1.1 Solvent: Most dyes are typically dissolved in organic solvents, such as anhydrous DMSO.
1.2 Concentration Recommendation: It is generally recommended to prepare a high-concentration stock solution of 1-10 mM.

2. Working Solution Preparation
2.1 Diluent: Modified Krebs-Henseleit solution.
2.2 Working Concentration: 2.5 μM.
2.3 Notes: Prepare and use immediately.

3. Staining Procedure
3.1 Sample Types:
3.1.1 Intact heart tissue: Langendorff-perfused intact guinea pig hearts.
3.1.2 Tissue sections: Guinea pig left ventricular tissue sections (350 μm thick).
3.2 Incubation Conditions: Incubate with 2.5 μM fura-4F AM working solution for 30 min at 36±1 °C via cyclic perfusion.
3.3 Washing Step: Information not provided; this section is omitted.

4. Control Group
Information not provided; this section is omitted.

5. Detection and Analysis
5.1 Instrument: EMCCD high-speed camera.
5.1.1 Excitation wavelength: 385 nm; emission wavelength is filtered through a multi-band filter with a passband for calcium dye signals.
5.2 Result Analysis:
5.2.1 Changes in fluorescence intensity: Fura-4F AM can record intracellular Ca²⁺ transient signals in isolated hearts and heart tissue sections. After treatment with Nifedipine, the CaT amplitude decreases, indicating that this dye can be used to monitor drug-induced Ca²⁺ dynamic changes.
5.2.2 This dye is used for Ca²⁺ optical imaging in whole hearts and tissue sections in literature, with no description of subcellular localization.

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