I am seeking assistance and guidance regarding the staining of MDMs with LysoSensor™ Yellow/Blue dextran, 10,000 MW, Anionic, Fixable. Specifically, I am interested in understanding if there is a recommended protocol for this staining procedure and the optimal staining duration for accurate pH calculations.
I have come across some articles suggesting a staining duration of 24 hours, but my results have been unexpected and unusual. I would greatly appreciate it if someone with experience in this area could provide me with advice, insights, or a reliable protocol to ensure the accuracy of pH measurements using LysoSensor™ Yellow/Blue dextran.
Your assistance and expertise would be invaluable to my research, and I am open to any guidance or recommendations you may have.
Thank you in advance for your time and assistance.
Best regards,
To stain cells or intracellular structures with LysoSensor™ Yellow/Blue Dextran, which is a pH indicator that accumulates in acidic compartments such as lysosomes, here is a basic protocol you can follow:
Materials Needed:
- LysoSensor™ Yellow/Blue Dextran, 10,000 MW, anionic, fixable
- Suitable cell culture medium
- Cells or tissue of interest
- Incubator with temperature and CO2 control
- Fluorescence microscope
Staining Steps:
It's important to note that the optimal staining duration may vary depending on cell type and culture conditions, so it's advisable to perform optimization experiments to achieve the best results in your system.
Keep in mind that the choice of concentration, staining duration, and culture conditions can influence the results, so optimization is essential. Also, look for specific LysoSensor™ staining protocols for your cell type or tissue if they are available in the literature or from your institution's resources.
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