NBD concentration used for cell imaging

Dear Yeoul,

the current protocol has been optimized specifically for cells in suspension, which will be subsequently analyzed using flow cytometry. To ensure accurate results, it is crucial to maintain the correct cell density in the cell suspension, with a minimum of 1 million cells per milliliter.

For a 3 mL volume of cell suspension, we typically recommend utilizing the following quantities: 80 nmol of NBD-PE, 40 nmol of NBD-PC, and 20 nmol of either NBD-PS or NBD-SM, all dissolved in 20 µmol of DMSO.

However, it is important to note that excessive amounts of NBD lipids can potentially harm the cells (NBD-lipids resemble a bit lyso-lipids). Therefore, we advise keeping the quantity of NBD lipids as low as possible with a minimum of 1 million cells per milliliter while still achieving accurate results. This will depend in part on your flow cytometer.

If you have any further questions or require additional clarification, please do not hesitate to reach out. Success with the experiments.

Best regards,

Thomas