Celltrack Deep Red is a fluorescent dye with a fluorescence signal that can be maintained for at least 72 h and has good stability. Celltrack Deep Red can be used for cell tracing and multi-generation cell movement tracking. Within a cell population, Celltrack Deep Red is only transferred to daughter cells and not to neighboring cells (Ex/Em = 630/650 nm) [1].
In Vitro, Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs). 1. Dye preparation Stock solution: Dissolve in DMSO to a final concentration of 1 mM (protect from light and store in aliquots). Working solution: Dilute the stock solution to a working concentration of 250 nM-1 μM in serum-free medium. Preheat the staining working solution to 37 °C. 2. Cell labeling 2.1 Staining protocol for cells in suspension 2.1.1 Harvest cells by centrifugation and aspirate the supernatant. Resuspend the cells gently in pre-warmed Celltrack Deep Red Working Solution. 2.1.2 Incubate 15-45 minutes under growth conditions appropriate for the particular cell type. 2.1.3 Centrifuge the cells and remove the the dye. 2.1.4 Change to fresh culture medium. 2.1.5 Fluorescence microscopy: Ex/Em = 630/650 nm. 2.2 Staining protocol for adherent cells 2.2.1 Remove culture media, and gently add the pre-warmed Celltrack Deep Red Working Solution. 2.2.2 Incubate 15-45 minutes under growth conditions appropriate for the particular cell type. 2.2.3 Remove the Celltrack Deep Red Working Solution. 2.2.4 Change to fresh culture medium. 2.2.5 Fluorescence microscopy: Ex/Em = 630/650 nm. Notes 1. Please adjust the Celltrack Deep Red working solution concentration, incubation temperature and time according to actual conditions. 2. To maintain normal cellular physiology and reduce potential artifacts, keep the dye concentration as low as possible. 3. Avoid using buffers containing amino or thiol groups.
References:
[1]. Wang Y, et al. A Granzyme B-Cleavable T Cell-Targeted Bispecific Cell Vesicle Connector for Reversing New-Onset Type 1 Diabetes. J Am Chem Soc. 2025 Feb 5;147(5):4167-4179.
In Vitro, Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs). 1. Dye preparation Stock solution: Dissolve in DMSO to a final concentration of 1 mM (protect from light and store in aliquots). Working solution: Dilute the stock solution to a working concentration of 250 nM-1 μM in serum-free medium. Preheat the staining working solution to 37 °C. 2. Cell labeling 2.1 Staining protocol for cells in suspension 2.1.1 Harvest cells by centrifugation and aspirate the supernatant. Resuspend the cells gently in pre-warmed Celltrack Deep Red Working Solution. 2.1.2 Incubate 15-45 minutes under growth conditions appropriate for the particular cell type. 2.1.3 Centrifuge the cells and remove the the dye. 2.1.4 Change to fresh culture medium. 2.1.5 Fluorescence microscopy: Ex/Em = 630/650 nm. 2.2 Staining protocol for adherent cells 2.2.1 Remove culture media, and gently add the pre-warmed Celltrack Deep Red Working Solution. 2.2.2 Incubate 15-45 minutes under growth conditions appropriate for the particular cell type. 2.2.3 Remove the Celltrack Deep Red Working Solution. 2.2.4 Change to fresh culture medium. 2.2.5 Fluorescence microscopy: Ex/Em = 630/650 nm. Notes 1. Please adjust the Celltrack Deep Red working solution concentration, incubation temperature and time according to actual conditions. 2. To maintain normal cellular physiology and reduce potential artifacts, keep the dye concentration as low as possible. 3. Avoid using buffers containing amino or thiol groups.
References:
[1]. Wang Y, et al. A Granzyme B-Cleavable T Cell-Targeted Bispecific Cell Vesicle Connector for Reversing New-Onset Type 1 Diabetes. J Am Chem Soc. 2025 Feb 5;147(5):4167-4179.