Large Field-of-View and Deep Tissue Optical microscopy Based on Parallel Wavefront Correction Algorithm

ZHAO Qi1 SHI Xin1 GONG Wei2 HU Lejia1 ZHENG Yao1 ZHU Xinpei2 SI Ke

(1.State Key Laboratory of Modern Optical Instrumentation, College of Optical Science and Engineering, Zhejiang University, Hangzhou, Zhejiang, China 310027)
(2.Center for Neuroscience, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China 310058)
【Knowledge Link】adaptive optics

【Abstract】As for deep tissues, the field of view of one-time correction in the widely used adaptive optics is limited and the refresh rate of a spatial light modulator or a deformable mirror is also limited. Therefore, it is difficult for them to satisfy the requirement of large field-of-view (FOV) rapid correction of wavefront distortion and thus that of high-speed imaging. A parallel wavefront correction method is proposed based on the conjugate adaptive optics correction system and the coherent optical adaptive technique. In this method, without increasing the number of refresh times of spatial light modulator, the large FOV of one-time correction can be realized by means of the parallel measurement of wavefront distortion of multiple guide stars, which provides a feasible reference solution for the high-speed and high-resolution imaging of deep tissues. The simulation results show that when 9 guiding stars are used, the effective FOV of one-time correction by the proposed method is about 4.7 times that of the conventional method for a thin scattering medium composed of 5 layers of random phasemasks, and 4.6 times that of the conventional method for 120-μm thick mouse brain tissue. Moreover, the proposed method can further improve the FOV of one-time correction by increasing the number of guide stars while the correction time does not significantly increase, which has broad application prospect in the large FOV imaging of in vivo biological tissues.

【Keywords】 imaging systems; microscopy; deep tissue imaging; scattering measurement; adaptive optics; wavefront shaping;


【Funds】 National Natural Science Foundation of China (31571110, 81771877) Natural Science Foundation of Zhejiang Province (LY16F050002, LZ17F050001)

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(Translated by LIU T)


    [1] Betzig E, Trautman J K. Near-field optics: microscopy, spectroscopy, and surface modification beyond the diffraction limit [J]. Science, 1992, 257 (5067): 189–195.

    [2] Helmchen F, Denk W. Deep tissue two-photon microscopy [J]. Nature Methods, 2005, 2 (12): 932–940.

    [3] Ahrens M B, Orger M B, Robson D N, et al. Whole-brain functional imaging at cellular resolution using light-sheet microscopy [J]. Nature Methods, 2013, 10 (5): 413–420.

    [4] Babcock H W. The possibility of compensating astronomical seeing [J]. Publications of the Astronomical Society of the Pacific, 1953, 65 (386): 229.

    [5] Rao C H, Jiang W H, Ling N, et al. Temporal correction effectiveness of adaptive optical system for light wave atmospheric propagation [J]. Acta Optica Sinica, 2001, 21 (8): 933–938 (in Chinese).

    [6] Jiang W H, Zhang Y D, Rao C H, et al. Progress on adaptive optics of institute of optics and electronics, Chinese Academy of Sciences [J]. Acta Optica Sinica, 2011, 31 (9): 0900106 (in Chinese).

    [7] Liang J Z, Williams D R, Miller D T. Supernormal vision and high-resolution retinal imaging through adaptive optics [J]. Journal of the Optical Society of America A, 1997, 14 (11): 2884–2892.

    [8] Tan Z J, Xie J, Lu J, et al. High spatial resolution confocal microscopy using adaptive optics [J]. Laser & Optoelectronics Progress, 2012, 49 (9): 090002 (in Chinese).

    [9] Booth M J. Adaptive optical microscopy: the ongoing quest for a perfect image [J]. Light: Science&Applications, 2014, 3 (4): e165.

    [10] Wang C, Ji N. Characterization and improvement of three-dimensional imaging performance of GRIN-lens-based two-photon fluorescence endomicroscopes with adaptive optics [J]. Optics Express, 2013, 21 (22): 27142–27154.

    [11] Wang K, Milkie D E, Saxena A, et al. Rapid adaptive optical recovery of optimal resolution over large volumes [J]. Nature Methods, 2014, 11 (6): 625–628.

    [12] Wang C, Liu R, Milkie D E, et al. Multiplexed aberration measurement for deep tissue imaging in vivo [J]. Nature Methods, 2014, 11 (10): 1037–1040.

    [13] Li J, Beaulieu D R, Paudel H, et al. Conjugate adaptive optics in widefield microscopy with an extended-source wavefront sensor [J]. Optica, 2015, 2 (8): 682–688.

    [14] Mertz J, Paudel H, Bifano T G. Field of view advantage of conjugate adaptive optics in microscopy applications [J]. Applied Optics, 2015, 54 (11): 3498–3506.

    [15] Park J H, Sun W, Cui M. High-resolution in vivo imaging of mouse brain through the intact skull [J]. Proceedings of the National Academy of Sciences, 2015, 112 (30): 9236–9241.

    [16] Wu T W, Cui M. Numerical study of multi-conjugate large areawavefront correction for deep tissue microscopy [J]. Optics Express, 2015, 23 (6): 7463–7470.

    [17] Cui M. Parallel wavefront optimization method for focusing light through random scattering media [J]. Optics Letters, 2011, 36 (6): 870–872.

    [18] Liu R, Milkie D E, Kerlin A, et al. Direct phase measurement in zonal wavefront reconstruction using multidither coherent optical adaptive technique [J]. Optics Express, 2014, 22 (2): 1619–1628.

    [19] Si K, Fiolka R, Cui M. Fluorescence imaging beyond the ballistic regime by ultrasound-pulse-guided digital phase conjugation [J]. Nature Photonics, 2012, 6 (10): 657–661.

    [20] Si K, Reto F, Cui M. Breaking the spatial resolution barrier via iterative sound-light interaction in deep tissue microscopy [J]. Scientific Reports, 2012, 2 (10): 748.

This Article


CN: 31-1339/TN

Vol 45, No. 12, Pages 224-231

December 2018


Article Outline



  • 1 Introduction
  • 2 Design of CAO correction system
  • 3 Principle of parallel wavefront correction algorithm
  • 4 Results and analysis
  • 5 Conclusions
  • References