[Application of two-photon excitation fluorescence imaging to real-time investigation of mouse preimplantation embryo].
The blue shift of a two-photon excitation absorption peak allows the application of a single wavelength to the simultaneous excitation of several fluorochromes with disparate emission characteristics. An output at 730 nm of a mode-locked femtosecond Ti-sapphire laser was used to excite four different commonly used fluorochromes, namely Hoechst 33342, Fluo-4, PI and Indo-1, and characteristic fluorescence images were obtained using (455 +/- 15) nm, (540 +/- 15)nm, (580 +/- 16) nm and (500 +/- 15) nm filters respectively. An approach to exciting with a single wavelength, staining with two different fluorochromes, and collecting fluorescence in two separate channels was employed to study mouse preimplantation embryos by 3D and 4D real-time imaging. Combined with the merits of two-photon excitation fluorescence imaging, such as better penetration, less photon-damage, and higher signal-to-noise ratio, this approach was supposed to be a novel multi-parameter investigating tool for mouse preimplantation embryos development study.
Cheng
Bing
B
The Key Laboratory of Atomic and Molecular Nanosciences, Ministry of Educatioin, Tsinghua University, Beijing 100084, China.
Lin
Dan-Ying
DY
Wang
Xiao-Guang
XG
Chen
Die-Yan
DY
Ma
Wan-Yun
WY
D000818:Animals; D003199:Computer Systems; D010064:Embryo Implantation; D004622:Embryo, Mammalian / Q000166:cytology; D005260:Female; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence / Q000379:methods*