Abstract

Two-photon excitation fluorescence (TPEF) microscopy has evolved into a versatile tool in biological research. However, the multiplexing capability of TPEF microscopy is limited by the narrow spectral bandwidth of the light source. In this study, we apply a photonic crystal fiber in TPEF microscopy to broaden the excitation source bandwidth. We tuned the spectral window using a spatial light modulator as a programmable diffraction grating that was placed behind a prism pair. In addition, we combined a grating pair to compensate for dispersion to improve the two-photon excitation efficiency. The combination of a broad spectrum and a programmable grating enabled fast spectral window tuning rate on a time scale of tens of milliseconds. We demonstrate the performance of our method by imaging live B16 cells labeled with four emission spectrum overlapped fluorescent proteins.

© 2023 Optica Publishing Group

Two-photon microscope using a fiber-based approach for supercontinuum generation and light delivery to a small-footprint optical head

Youbo Zhao, Gopi Maguluri, R. Daniel Ferguson, Haohua Tu, Kush Paul, Stephen A. Boppart, Daniel A. Llano, and Nicusor Iftimia
Opt. Lett. 45(4) 909-912 (2020)

Rapid coherent Raman hyperspectral imaging based on delay-spectral focusing dual-comb method and deep learning algorithm

Yujia Zhang, Minjian Lu, Jiaqi Hu, Yan Li, Perry Ping Shum, Jinna Chen, and Haoyun Wei
Opt. Lett. 48(3) 550-553 (2023)

Time-resolved detection enables standard two-photon fluorescence microscopy for in vivo label-free imaging of microvasculature in tissue

Dong Li, Wei Zheng, Wei Zhang, Seng Khoon Teh, Yan Zeng, Yi Luo, and Jianan Y. Qu
Opt. Lett. 36(14) 2638-2640 (2011)

Rotated chirped volume Bragg gratings for compact spectral analysis

Oussama Mhibik, Murat Yessenov, Lam Mach, Leonid Glebov, Ayman F. Abouraddy, and Ivan Divliansky
Opt. Lett. 48(5) 1180-1183 (2023)

Tunable dissipative soliton Tm-doped fiber laser operating from 1700 nm to 1900 nm

Xinyang Liu, Jayanta K. Sahu, and Regina Gumenyuk
Opt. Lett. 48(3) 612-615 (2023)