TPE-SQ5

Operating Instructions (The following is a recommended experimental protocol for guidance only; adjust according to specific requirements)

1. Stock Solution Preparation
1.1 Solvents: THF; or water adjusted to pH 9.
1.2 Recommended Concentrations: 10 μM; or 30 μM.

2. Working Solution Preparation
2.1 Diluents: THF/H₂O mixture (water volume fraction = 95%), cell culture medium; or distilled water/tap water.
2.2 Working Concentrations: 5-10 μM for cell imaging; or 15 μM for water sample analysis.
2.3 Notes: Adjust the working solution concentration as needed; prepare and use immediately.

3. Staining Procedures
3.1 Sample Type Description
3.1.1 Adherent cells^[1]: Hela cells, 3T3 cells, MCF-7 cells.
3.1.2 Suspension cells^[1]: RAW264.7 cells.
3.1.3 Water samples^[2]: Tap water and distilled water samples.
3.2 Incubation Conditions
3.2.1 Adherent cells^[1]: Co-incubate with 10 μM TPE-SQ5 nanoparticles at 37°C for 30 min to 2 h; for detection of exogenous ClO^-, an additional co-incubation with 100 μM ClO^- for 30 min is required.
3.2.2 Suspension cells^[1]: Co-incubate with 5 μM TPE-SQ5 nanoparticles for 2 h; for detection of endogenous ClO^-, pretreat cells with LPS (1 μg mL^-1) for 12 h first, then incubate with PMA (1 μg mL^-1) for 30 min before adding TPE-SQ5 nanoparticles.
3.2.3 Water samples^[2]: No specific temperature or time requirements; no light avoidance requirement.
3.3 Washing Step: No specified washing step.

4. Control Setup
4.1 Negative Controls:
4.1.1 Hela cells incubated only with TPE-SQ5 nanoparticles.
4.1.2 RAW264.7 cells incubated only with TPE-SQ5 nanoparticles.
4.2 Positive control for verifying the experimental system.
4.3 Blank control for eliminating reagent fluorescence interference.

5. Detection and Analysis
5.1 Instrument Types: Fluorescence spectrophotometer, laser confocal scanning microscope.
5.2 Excitation/Emission Wavelengths
5.2.1 Fluorescence spectrophotometer:
5.2.1.1 In vitro cell-related detection^[1]: Ex = 380 nm, Em = 485 nm and 643 nm.
5.2.2 Laser confocal scanning microscope^[1]:
5.2.2.1 Red channel: Ex = 488 nm, Em = 550-650 nm.
5.2.2.2 Green channel: Ex = 405 nm, Em = 450-550 nm.
5.3 Result Analysis
5.3.1 Fluorescence Intensity Changes:
5.3.1.1 In vitro cell-related detection^[1]: The fluorescence intensity ratio at 485 nm and 643 nm (F₄₈₅/F₆₄₃) increases with the rise of ClO^- concentration; when ClO^- exists in cells, the fluorescence of the red channel weakens while that of the green channel enhances.
5.3.2 Fluorescence Localization^[1]: TPE-SQ5 nanoparticles localize inside cells.
5.3.3 Color Changes:
5.3.3.1 In vitro cell-related detection^[1]: After adding ClO^-, the emission light color changes from deep red to blue-green; when the intracellular ClO^- level increases, the fluorescence shifts from red to green.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.