Axial gradient excitation accelerates volumetric imaging of two-photon microscopy

被引：12

作者：

Gao, Yufeng ^{[1
,2
]}

Xia, Xianyuan ^{[1
,2
]}

Liu, Lina ^{[1
,2
]}

Wu, Ting ^{[1
,2
]}

Chen, Tingai ^{[1
,2
]}

Yu, Jia ^{[1
,2
]}

Xu, Zhili ^{[3
]}

Wang, Liang ^{[1
,2
]}

Yan, Fei ^{[3
]}

Du, Zhuo ^{[4
,5
]}

Chu, Jun ^{[1
,2
]}

Zhan, Yang ^{[6
]}

Peng, Bo ^{[7
]}

Li, Hui ^{[1
,2
]}

Zheng, Wei ^{[1
,2
]}

机构：

[1] Chinese Acad Sci, Shenzhen Inst Adv Technol, Guangdong Prov Key Lab Biomed Opt Imaging Technol, Shenzhen Key Lab Mol Imaging,Res Ctr Biomed Opt &, Shenzhen 518055, Peoples R China

[2] Chinese Acad Sci, Shenzhen Inst Adv Technol, CAS Key Lab Hlth Informat, Shenzhen 518055, Peoples R China

[3] Chinese Acad Sci, Shenzhen Inst Adv Technol, Inst Biomed & Hlth Engn, Paul C Lauterbur Res Ctr Biomed Imaging, Shenzhen 518055, Peoples R China

[4] Chinese Acad Sci, Inst Genet & Dev Biol, State Key Lab Mol Dev Biol, Beijing 100101, Peoples R China

[5] Chinese Acad Sci, Inst Genet & Dev Biol, Key Lab Genet Network Biol, Beijing 100101, Peoples R China

[6] Chinese Acad Sci, Shenzhen Inst Adv Technol, Brain Cognit & Brain Dis Inst, Shenzhen 518055, Peoples R China

[7] Chinese Acad Sci, Shenzhen Inst Adv Technol, Ctr Micro Nano Syst & Bion Med, Shenzhen 518055, Peoples R China

来源：

PHOTONICS RESEARCH | 2022年 / 10卷 / 03期

基金：

中国国家自然科学基金;

关键词：

ADAPTIVE OPTICS; ELECTROMAGNETIC DIFFRACTION; MULTIPHOTON MICROSCOPY; NEURONAL-ACTIVITY; FIELD; DEPTH; SEGMENTATION; ILLUMINATION; ALGORITHM; SYSTEMS;

D O I：

10.1364/PRJ.441778

中图分类号：

O43 [光学];

学科分类号：

070207 ; 0803 ;

摘要：

Two-photon excitation fluorescence microscopy (TPM), owing to its capacity for subcellular resolution, intrinsic optical sectioning, and superior penetration depth in turbid samples, has revolutionized biomedical research. However, its layer-by-layer scanning to form a three-dimensional image inherently limits the volumetric imaging speed and increases phototoxicity significantly. In this study, we develop a gradient excitation technique to accelerate TPM volumetric imaging. The axial positions of the fluorophores can be decoded from the intensity ratio of the paired images obtained by sequentially exciting the specimen with two axially elongated two-photon beams of complementary gradient intensities. We achieved a 0.63 mu m axial localization precision and demonstrate the flexibility of the gradient TPM on various sparsely labeled samples, including bead phantoms, mouse brain tissues, and live macrophages. Compared with traditional TPM, our technique improves volumetric imaging speed (by at least sixfold), decreases photobleaching (i.e., less than 2.07 +/- 2.89% in 25 min), and minimizes photodamages. (C) 2022 Chinese Laser Press

引用

页码：687 / 696

页数：10