GLF16 FA is a hydrophilic fluorescent SBB analogue for rapid detection of senescent cells using flow cytometry and live cell sorting.

Instructions
I. Solution preparation
1. Stock solution: Dissolve GLF16 FA in an appropriate solvent, usually anhydrous DMSO or PBS, to prepare a stock solution with a concentration of 1–10 mM.
2. Working solution: Dilute the stock solution to a working concentration, usually 1–5 µM, using an experimental buffer (such as PBS, pH 7.4) or culture medium.
II. Operation steps
Cell culture experiment
(1) Cell type: GLF16 FA can be used to detect senescent cells, especially for a variety of cell lines cultured in vitro.
(2) Treatment method: Add GLF16 FA to the cell culture medium and culture the cells for 30 minutes to 1 hour at 37°C, away from light.
(3) Cell function detection:
a. Senescence marker detection: Detect the fluorescence intensity of GLF16 FA in cells by flow cytometry or live cell sorting to identify senescent cells.
b. Cell survival rate detection: CCK-8 or other cell activity detection methods can be used to evaluate cell survival rate.
c. Senescence marker detection: Combined with other senescence markers, such as SA-β-gal, the characteristics of senescent cells can be further verified.
Animal experiments
(1) Treatment method: GLF16 FA is administered to experimental animals by intraperitoneal injection or tail vein injection. The dose is usually 1–10 mg/kg depending on the experimental design.
(2) Senescence detection: After injection, the fluorescence signal of senescent cells in vivo is detected by flow cytometry or other imaging techniques.
3. Calibration and control
(1) Control group: Set up an untreated cell or animal group as a control to verify the effect of GLF16 FA.
(2) Standard curve: Use senescent cell samples with known concentrations to establish a standard curve between the fluorescence signal and the proportion of senescent cells.
Notes
(1) Storage conditions: GLF16 FA should be stored at -20°C in the dark and avoid repeated freezing and thawing.
(2) Fluorescence detection: The excitation wavelength is usually 485–495 nm and the emission wavelength is 515–525 nm.
(3) Photosensitivity: GLF16 FA is light-sensitive and strong light exposure should be avoided during the experiment.
(4) Solubility: Ensure that GLF16 FA is completely dissolved during dissolution to avoid any solid residue.