Cholyl-L-lysine fluorescein disodium

Guide (The following is our recommended protocol, which serves only as a guideline and should be modified according to your specific needs)^[1].
1. Centrifuge organoids at 300 g for 2 minutes, then remove the supernatant.
2. Add 100 μL of medium containing the corresponding fluorescent substrate Cholyl-L-lysine fluorescein disodium, with or without inhibitors, to each sample, and incubate at 37 °C until the specified time point.
33 After incubation, add 1 mL of ice-cold PBS to terminate substrate transport, and centrifuge at 300 rpm for 2 minutes.
4. Remove the supernatant, wash the sample with 1 mL of PBS, and repeat this step 3 times. Seed the obtained organoids into a 24-well plate containing 300 μL of pre-warmed HCM medium at 37 °C, and perform analysis using an inverted fluorescence microscope and CellSens software.
5. For semi-quantitative analysis, capture at least 7 images containing at least 7 organoids each under the brightfield channel. Subsequently, capture corresponding fluorescence images of each channel using the same fluorescence settings. Use ImageJ software to mark organoids in the brightfield images and calculate their area. Next, set an intensity threshold for the fluorescence images and keep it unchanged throughout the analysis. Subsequently, use ImageJ to determine the fluorescence-positive area and calculate the ratio of the fluorescence-positive area to the total area of organoids.

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