Description: Deep Red Fluorescent acidotropic probes for labeling and tracking acidic organelles in live cells Form: Liquid Directions for Use : 1. Prepare the working solution Add Lysosome Red Probes to the cell culture medium at the ratio of 1:10,000 - 1:20,000. The final concentration is 50-100 nM. The working solution can be warmed at 37°C before use. 2. Label the lysosomes For adherent cells, discard the cell medium and wash with 1 x PBS. Adding the working solution, and incubate the cells at 37°C for 30 mins to 2 hours. Discard the working solution, wash with 1 x PBS for three times, and then photographed under a fluorescence microscope. For suspension cells, centrifuge to obtain a cell pellet and aspirate the supernatant. Resuspend the cells gently in probe-containing medium, and incubate the cells at 37°C for 30 mins to 2 hours. Centrifuge to obtain a cell pellet and aspirate the supernatant, wash with 1 x PBS for three times, and then photographed under a fluorescence microscope. Notice 1. If the staining effect is not good, the concentration of the probe in the working fluid can increase the concentration of the working solution , or can extend the reaction time. 2. In order to reduce the background, pleasee use lower concentration probes. 3. Take photos quickly, because the dye is easy to quench.Application : The Lysosome Deep Red Probes, which consist of a fluorophore linked to a weak base that is only partially protonated at neutral pH, are freely permeant to cell membranes and typically concentrate in spherical organelles. Their mechanism of retention has not been firmly established but is likely to involve protonation and retention in the membranes of the organelles, although staining is generally not reversed by subsequent treatment of the cells with weakly basic cell-permeant compounds. Note that in Lysosome Deep Red Probes stained cells, the lysosomal fluorescence may constitute only a small portion of total cellular fluorescence, making it difficult to quantitate the number of lysosomes by flow cytometry or fluorometry.Storage/Stability : Store at -20 °C in the dark for 6 months. Platform : Ex/Em = 649/665 nm