[Two-photon excitation fluorescence of 5-ALA induced PpIX in DHL cells].
Two-photon fluorescence microscopy is a novel imaging technique, which is primarily sensitive to a specimen's response coming from an in-focus plane, thus has low photo-bleaching and photo-damage to biological samples. 5-ALA induced production of PpIX in DHL cells was excited by 820 nm femtosecond laser; two-photon excitation fluorescence of single cell was obtained in Lambda mode of laser scanning confocal microscope. The specific fluorescence intensity of PpIX which accumulated in DHL cells was measured at 2, 4 and 10 mmol x L(-1) concentration of 5-ALA with different incubation time, which reflected the kinetics of 5-ALA accumulated in DHL cells. Accumulation of PpIX in DHL cells was a dynamic change process. Biphasic alterations of PpIX accumulation were noted: PpIX content enhanced with the increasing time and reached the maximal value around 3 h, however PpIX content decreased in the subsequent incubation time. Results indicate that two-photon fluorescence based on laser scanning microscope can be a useful technology for studying the kinetics of 5-ALA induced PpIX production in DHL cells and other leukemia cells.
Huang
Zu-Fang
ZF
Key Laboratory of Optoelectronic Science and Technology for Medicine, Ministry of Education, Fujian Normal University, Fuzhou 350007, China.
Chen
Rong
R
Li
Yong-Zeng
YZ
Chen
Guan-Nan
GN
Chen
Xian-Ling
XL
Feng
Shang-Yuan
SY
Jia
Pei-Min
PM
D000622:Aminolevulinic Acid / Q000494:pharmacology*; D045744:Cell Line, Tumor; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007700:Kinetics; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D017785:Photons; D011524:Protoporphyrins / Q000378:metabolism*
D011524:Protoporphyrins; D000622:Aminolevulinic Acid; C028025:protoporphyrin IX