Photoswitchable PM8

Guide (The following is a recommended experimental protocol for reference only and should be adjusted according to specific requirements)
1. Working Solution Preparation
1.1 Diluent: PBS
1.2 Working concentration: 1 μM
1.3 Notes: Adjust the working solution concentration as needed; prepare fresh before use.

2. Staining Procedure
2.1 Sample type: Mammalian cells (HEK293, U2OS)^[1]
2.1.1 Adherent cells
2.2 Incubation conditions: Incubate with 1 μM dye working solution for 10 min at room temperature; image directly without washing

3. Detection and Analysis
3.1 Instruments: Confocal fluorescence microscope; Single-molecule localization microscopy (PALM)
3.1.1 Confocal imaging excitation wavelength: 488 nm
35.1.2 PALM imaging: Activation with 405 nm laser, excitation with 561 nm laser
3.1.3 Emission wavelength: 628 nm (for Z isomer)
3.2 Result analysis:
3.2.1 Fluorescence is selectively activated in hydrophobic cytoplasmic membrane environments, exhibiting a high signal-to-noise ratio
3.2.2 Fluorescence localizes to the cytoplasmic membrane, enabling visualization of filopodia
3.2.3 The average localization precision of super-resolution PALM imaging is 17 nm for fixed cells and 25 nm for live cells, resolving fine membrane structures that cannot be captured by diffraction-limited imaging.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. .