Kinetics and subcellular localization of 5-ALA-induced PpIX in DHL cells via two-photon excitation fluorescence microscopy.
Two-photon excitation fluorescence (TPEF) microscopy was used to measure the 5-aminolevulinic acid (5-ALA)-induced PpIX fluorescence in follicular lymphoma DHL cells. Kinetics of 5-ALA-induced PpIX accumulation in DHL cells under various 5-ALA concentrations was studied. We found that during the course of continuous incubation with 5-ALA, the relationship between the DHL cell fluorescence signal and the incubation time showed a biphasic variation. Initially the PpIX signal increased with the incubation time and reached the maximal value at about 3 h, and then it decreased with time during the subsequent incubation period. By labeling the 5-ALA incubated DHL cells with different organelle-specific fluorescence probes: Rhodamine 123 (for mitochondria), DioC6(3) (for endoplasmic reticulum) and LysoTracker Green (for lysosomes) respectively, we found that 5-ALA-induced PpIX was primarily localized in endoplasmic reticulum and mitochondria; its concentration in the lysosome was much lower. The results suggested that 5-ALA could potentially be an effective photosensitizer in photodynamic purging of DHL cells. Two-photon excitation fluorescence microscope is a useful tool for studying 5-ALA-induced PpIX subcellular localization.
Chen
Rong
R
Key Laboratory of Optoelectronic Science and Technology for Medicine, Ministry of Education, Fujian Normal University, Fuzhou, Fujian 350007, PR China.
Huang
Zufang
Z
Chen
Guannan
G
Li
Yongzeng
Y
Chen
Xianlian
X
Chen
Jianxin
J
Zeng
Haishan
H
D000622:Aminolevulinic Acid / Q000494:pharmacology*; D045744:Cell Line, Tumor; D006801:Humans; D008224:Lymphoma, Follicular / Q000737:chemistry*; D008856:Microscopy, Fluorescence / Q000379:methods*; D010778:Photochemotherapy; D011524:Protoporphyrins / Q000032:analysis*
D011524:Protoporphyrins; D000622:Aminolevulinic Acid; C028025:protoporphyrin IX