LDs-BCA

Guidelines (The recommended experimental protocol below is for guidance only and should be modified according to your specific requirements)

1. Stock Solution Preparation
1.1 Solvent: Ultrapure water containing 1% DMSO.
1.2 Concentration Recommendation: Prepare a high-concentration stock solution with a concentration of 1-10 mM.

2. Working Solution Preparation
2.1 Diluent: Serum-free medium or PBS is commonly used. Proteins and esterases in serum may interfere with staining results or cause dye hydrolysis.
2.2 Working Concentration: 10 μM.
2.3 Notes: Adjust the working solution concentration as needed; prepare and use it immediately.

3. Staining Procedures
3.1 Adherent Cell Assay^[1]:
3.1.1 Sample Types: A549 cells, HeLa cells.
3.1.2 Incubation Conditions: Incubate with 10 μM LDs-BCA for 10 min.
3.2 In Vivo Mouse Gastric Tissue Assay^[2][3]:
3.2.1 Sample Types: Gastric tissues of normal and obese live mice.
3.2.2 Incubation Conditions: Incubate with LDs-BCA; the observable duration of fluorescence signal reaches up to 12 h; monitor lipid droplet metabolism in gastric tissues.
3.3 In Vivo Live Mouse Assay^[1]:
3.3.1 Sample Types: Obese and normal live mice.
3.3.2 Incubation Conditions: Intraperitoneally inject 100 μL of 1 mM LDs-BCA; perform imaging 10 min later.

4. Control Setup
4.1 Cell Assay Controls: Untreated HeLa cells, HeLa cells incubated with complete 1640 medium, untreated A549 cells.
4.2 In Vivo Mouse Assay Controls: Normal mice injected with 1 mM LDs-BCA, obese mice without probe injection, gastric tissues of normal mice (negative control), gastric tissues of obese mice (positive control).
4.3 System Validation Controls: Positive control (for validating the experimental system); blank control (for eliminating reagent fluorescence interference).

5. Detection and Analysis
5.1 Instrument Types: Laser confocal scanning microscope; fluorescence imaging system.
5.2 Excitation/Emission Wavelengths:
5.2.1 Cell Imaging: Ex = 565 nm, Em = 620-730 nm.
5.2.2 In Vivo Imaging: Ex = 560 nm, Em = 670 nm; Ex = 570 nm, Em = 650 nm.
5.3 Result Analysis:
5.3.1 Fluorescence Intensity Evaluation: The fluorescence intensity in the gastric region of obese mice is significantly higher than that of normal mice; the fluorescence intensity of HeLa cells treated with oleic acid or HBSS is stronger than that of control cells; strong fluorescence signals are observable in gastric tissues of obese mice, while signals in gastric tissues of normal mice are negligible.
5.3.2 Fluorescence Localization: Red fluorescence specifically localizes to intracellular lipid droplets (with a high overlap coefficient with commercial lipid droplet dyes); in in vivo assays, red fluorescence targets gastric fat in obese mice.
5.3.3 Dynamic Monitoring: Track changes in the gastric fat microenvironment; monitor the dynamics of lipid droplet metabolism.
5.3.4 Fluorescence Color: LDs-BCA emits red fluorescence in lipid droplets and gastric fat.

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