Technical Reference #1922

1922.

Enhanced photodynamic efficacy towards melanoma cells by encapsulation of Pc4 in silica nanoparticles Baozhong Zhao; Jun-Jie Yin; Piotr J. Bilski; Colin F. Chignell; Joan E. Roberts; Yu-Ying He, National Institute of Environmental Health Sciences, Toxicology and Applied Pharmacology, 241(1922), (2009)

Keywords:
Photodynamic therapy; Nanoparticles; Melanoma; Phthalocyanine; Singlet oxygen; Apoptosis

Materials & Methods:
Fluorescence confocal imaging. We employed fluorescence microscopy to study cellular uptake and subcellular localization. A375 and B16F10 cells were seeded into 35 mm dishes containing a glass coverslip-covered 14 mm cutout (MatTek Ashland MA) for live cell microscopy measurement. Cells were incubated with 2 μM Pc4 or Pc4SNP for 2 h at 37 °C. For measurement of subcellular localization cells were further stained with 100 nM MitoTracker Green FM (Invitrogen/Molecular Probes Eugene OR) which is a specific fluorescence dye for mitochondria or 60 nM LysoTraker Green DND-26 (Invitrogen/Molecular Probes Eugene OR) which is a specific fluorescence dye for lysosomes. Confocal fluorescence imaging was performed with a Zeiss LSM-510 META confocal microscope. For Pc4 fluorescence excitation was carried out at 633 nm and emission was collected for wavelengths greater than 650 nm. For MitoTracker Green FM or LysoTracker Green DND-26 fluorescence excitation was carried out at 488 nm and emission was collected for wavelengths between 505 and 550 nm.

Microscopic Technique
Fluorescence Microscopy

Cell Type(s)
Pigmented mouse melanoma B16F10 cells