Cell-Tracker Red CMTPX (Synonyms: CellTracker Red CMTPX Dye)

1. Cell preparation

Cultivate cells in appropriate culture medium. Adhering cells can climb and grow in a culture dish containing a cover glass, and sufficient growth medium can be loaded.

The following steps describe the process of adding dye to cultured cells and imaging them under a fluorescence microscope. Various factors, such as loading dyes into cells or tissues, may require modifications to certain conditions based on specific cell types.

The optimal staining concentration of the probe needs to be adjusted according to its purpose. It is recommended to measure at least one concentration within a 10 fold range when conducting experiments. Generally speaking, long-term staining (>3 days) or the use of rapidly dividing cells require 5-25 uM of dye. For short-term experiments such as vitality testing, use low concentration dyes (0.5-5uM). To maintain normal cell morphology and reduce potential artifacts, use low concentrations of dyes as much as possible.

2.1 Preparation of Cell Tracker staining solution

① Before opening the bottle, remove the product from the refrigerator and place it in Xingwen to let it warm up for at least 20 minutes

② Dissolve the powder with high-quality anhydrous DMSO to a concentration of 10mM. For example, for 50ug Cell Tracker Red CMTPX (Mw: 686.25 g/mol) freeze-dried powder, add 7.2 μ IDMSO is fully dissolved to obtain 10mM mother liquor.

③ Dilute the mother liquor with serum-free culture medium to a working concentration of 0.5-25uM. Preheat the dyeing solution to 37 ℃.

2.2 Suspension cell staining steps

① Collect cells by centrifugation and aspirate the supernatant. Gently resuspend cells using preheated Cell Tracker staining solution.

② Incubate for 15-45 minutes under suitable growth conditions for specific cell types.

③ Centrifuge the cells and aspirate the Cell Tracker staining solution.

④ Add the selected culture medium and distribute the labeled cells onto a glass slide or into the selected culture vessel.

⑤ Select appropriate excitation and emission wavelength filters according to Appendix 1 for imaging detection.

2.3 Staining steps for adherent cells

① Remove the culture medium.

② Gently add preheated Cell Tracker staining solution.

③ Incubate for 15-45 minutes under suitable growth conditions for specific cell types.

④ Suck off the Cell Tracker staining solution.

⑤ Add the selected culture medium.

⑥ Select appropriate excitation and emission wavelength filters according to Table 1 for imaging detection.

3. Fluorescence microscopy observation

Cell Tracker fluorescent probes can be detected using various fall emission fluorescence optical microscopes with standard optics and image enhancement. Select the appropriate filter according to the dye. See Appendix 1 for the spectral characteristics of the Cel Tracker fluorescent probe.

matters needing attention

Fluorescent dyes all have quenching problems, so be careful to avoid light during storage and operation. Avoid using buffer solutions containing hydrogen and organic groups. For your safety and health, please wear laboratory clothes and disposable gloves when operating.