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基于单一分光棱镜干涉仪的双通路定量相位显微术

孙腾飞 卢鹏 卓壮 张文浩 卢景琦

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基于单一分光棱镜干涉仪的双通路定量相位显微术

孙腾飞, 卢鹏, 卓壮, 张文浩, 卢景琦

Dual-channel quantitative phase microscopy based on a single cube beamsplitter interferometer

Sun Teng-Fei, Lu Peng, Zhuo Zhuang, Zhang Wen-Hao, Lu Jing-Qi
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  • 仅仅使用一个单独的分光棱镜(BS),实现了一种用于生物细胞三维成像的双通路定量相位显微术.不同于传统的使用方法,将BS倾斜放置,使中央半反射层与入射光光轴之间存在一个非常小的角度.这样基于BS的分光特性,经过BS后的透射光束和反射光束将会叠加在一起并形成干涉.调节样品位置,利用相机拍摄同时获得了存在π相移的双通路干涉图.这种离轴干涉模式,只需要记录单幅干涉图就可以获得真实的相位信息,方法结构简单,易于操作,适用于微小透明样品的三维形貌测量.
    Quantitative phase microscopy, as a non-destructive and non-invasive measurement technique, can indirectly reflect three-dimensional (3D) morphology and optical properties of transparent microstructure object by measuring phase information. In recent years, this kind of technique has been widely used to detect and investigate the characteristics of biological cells and it has become more and more important in the field of modern biomedical and life science. In this paper, only by using a single cube beamsplitter interferometer, a simple single-shot dual-channel quantitative phase microscopic measurement technique is demonstrated for 3D quantitative phase imaging of biological cells. In the proposed method, a conventional non-polarized cube beamsplitter is the most pivotal element. Unlike its traditional application method, the cube beamsplitter is tilted in a nonconventional configuration and the illumination beam is only incident on the left (or right) half of the cube beamsplitter (just the one side of central semi-reflecting layer), and a very small angle is introduced between the central semi-reflecting layer and the optical axis of incident beam. Based on the light splitting characteristic of the cube beamsplitter, two replicas of incident beam are generated. These two generated replicas (transmission beam and reflection beam) are of symmetry with respect to each other, and they will encounter and form interference when the direction of the incident beam meets a certain condition. Adjust the sample to a suitable position and make it only contact one half of incident beam, and the modulated beam will be seen as the object beam and the remaining clean half of incident beam as the reference beam. When the interference phenomenon occurs, two interference channels with a relative π (rad) phase-shift in one interferogram are acquired simultaneously only using one digital camera, and the higher spatial frequency of interference fringes can be achieved by adjusting a relatively big angle between the central semi-reflecting layer and the optical axis of incident beam. Because of the off-axis interference mode, we only need to record one interferogram to gain the continuous phase information and avoid using complex phase-shift techniques. At the same time, this proposed method is of simple structure and easy to operate due to using less ordinary off-the-shelf optical elements. All these simplify the structure of the system and reduce the cost of the system as much as possible. Finally, the phase information of paramecium is successfully obtained from different interference channels respectively. Furthermore, according to the characteristic of π (rad) phase-shift, we also realize the calibration and determination of ultimate precise phase information of sample by using the method of averaging between these two channels. The experimental results show that our proposed method is suitable for 3D surface morphology measurement of small transparent samples.
      通信作者: 卢景琦, Lu618@sdu.edu.cn
    • 基金项目: 山东省自然科学杰出青年基金(批准号:2013JQE27056)和山东省重点研发计划(批准号:2016GSF121023)资助的课题.
      Corresponding author: Lu Jing-Qi, Lu618@sdu.edu.cn
    • Funds: Project supported by the Shandong Provincial Natural Science Foundation for Distinguished Young Scholars, China (Grant No. 2013JQE27056) and the Shandong Provincial Key Research and Development Program, China (Grant No. 2016GSF121023).
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    Marquet P, Depeursinge C, Magistretti P J 2014 Neurophotonics 1 020901

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    Marquet P, Rothenfusser K, Rappaz B, Depeursinge C, Jourdain P, Magistretti P J 2016 Proc. SPIE 9718 97180K

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    Wang H Y, Liu F F, Liao W, Song X F, Yu M J, Liu Z Q 2013 Acta Phys. Sin. 62 054208 (in Chinese) [王华英, 刘飞飞, 廖薇, 宋修法, 于梦杰, 刘佐强 2013 物理学报 62 054208]

    [6]

    Li J C 2012 Acta Phys. Sin. 61 134203 (in Chinese) [李俊昌 2012 物理学报 61 134203]

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    Mir M, Tangella K, Popescu G 2011 Biomed. Opt. Express 2 3259

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    Shaked N T 2012 Opt. Lett. 37 2016

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    Anand A, Faridian A, Chhaniwal V, Mahajan S, Trivedi V, Dubey S K, Pedrini G, Osten W, Javidi B 2014 Appl. Phys. Lett. 104 103705

    [10]

    Mahajan S, Trivedi V, Vora P, Chhaniwal V, Javidi B, Anand A 2015 Opt. Lett. 40 3743

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    Coquoz S, Nahas A, Sison M, Lopez A, Lasser T 2016 J. Biomed. Opt. 21 126019

    [12]

    Di J L, Li Y, Xie M, Zhang J W, Ma C J, Xi T L, Li E P, Zhao J L 2016 Appl. Opt. 55 7287

    [13]

    Ma C J, Li Y, Zhang J W, Li P, Xi T L, Di J L, Zhao J L 2017 Opt. Express 25 13659

    [14]

    Zhang J W, Dai S Q, Ma C J, Di J L, Zhao J L 2017 Appl. Opt. 56 3223

    [15]

    Chhaniwal V, Singh A S G, Leitgeb R A, Javidi B, Anand A 2012 Opt. Lett. 37 5127

    [16]

    Yuan F, Yuan C J, Nie S P, Zhu Z Q, Ma Q Y, Li Y, Zhu W Y, Feng S T 2014 Acta Phys. Sin. 63 104207 (in Chinese) [袁飞, 袁操今, 聂守平, 朱竹青, 马青玉, 李莹, 朱文艳, 冯少彤 2014 物理学报 63 104207]

    [17]

    Singh A S G, Anand A, Leitgeb R A, Javidi B 2012 Opt. Express 20 23617

    [18]

    Lue N, Kang J W, Hillman T R, Dasari R R, Yaqoob Z 2012 Appl. Phys. Lett. 101 084101

    [19]

    Qu W J, Bhattacharya K, Choo C O, Yu Y J, Asundi A 2009 Appl. Opt. 48 2778

    [20]

    Gabai H, Shaked N T 2012 Opt. Express 20 26906

    [21]

    Lan B, Feng G Y, Zhang T, Liang J C, Zhou S H 2017 Acta Phys. Sin. 66 069501 (in Chinese) [兰斌, 冯国英, 张涛, 梁井川, 周寿桓 2017 物理学报 66 069501]

    [22]

    Takeda M, Ina H, Kobayashi S 1982 J. Opt. Soc. Am. 72 156

    [23]

    Cuche E, Marquet P, Depeursinge C 2000 Appl. Opt. 39 4070

  • [1]

    Li J C, Lou Y L, Gui J B, Peng Z J, Song Q H 2013 Acta Phys. Sin. 62 124203 (in Chinese) [李俊昌, 楼宇丽, 桂进斌, 彭祖杰, 宋庆和 2013 物理学报 62 124203]

    [2]

    Wang H Y, Zhang Z H, Liao W, Song X F, Guo Z J, Liu F F 2012 Acta Phys. Sin. 61 044208 (in Chinese) [王华英, 张志会, 廖薇, 宋修法, 郭中甲, 刘飞飞 2012 物理学报 61 044208]

    [3]

    Marquet P, Depeursinge C, Magistretti P J 2014 Neurophotonics 1 020901

    [4]

    Marquet P, Rothenfusser K, Rappaz B, Depeursinge C, Jourdain P, Magistretti P J 2016 Proc. SPIE 9718 97180K

    [5]

    Wang H Y, Liu F F, Liao W, Song X F, Yu M J, Liu Z Q 2013 Acta Phys. Sin. 62 054208 (in Chinese) [王华英, 刘飞飞, 廖薇, 宋修法, 于梦杰, 刘佐强 2013 物理学报 62 054208]

    [6]

    Li J C 2012 Acta Phys. Sin. 61 134203 (in Chinese) [李俊昌 2012 物理学报 61 134203]

    [7]

    Mir M, Tangella K, Popescu G 2011 Biomed. Opt. Express 2 3259

    [8]

    Shaked N T 2012 Opt. Lett. 37 2016

    [9]

    Anand A, Faridian A, Chhaniwal V, Mahajan S, Trivedi V, Dubey S K, Pedrini G, Osten W, Javidi B 2014 Appl. Phys. Lett. 104 103705

    [10]

    Mahajan S, Trivedi V, Vora P, Chhaniwal V, Javidi B, Anand A 2015 Opt. Lett. 40 3743

    [11]

    Coquoz S, Nahas A, Sison M, Lopez A, Lasser T 2016 J. Biomed. Opt. 21 126019

    [12]

    Di J L, Li Y, Xie M, Zhang J W, Ma C J, Xi T L, Li E P, Zhao J L 2016 Appl. Opt. 55 7287

    [13]

    Ma C J, Li Y, Zhang J W, Li P, Xi T L, Di J L, Zhao J L 2017 Opt. Express 25 13659

    [14]

    Zhang J W, Dai S Q, Ma C J, Di J L, Zhao J L 2017 Appl. Opt. 56 3223

    [15]

    Chhaniwal V, Singh A S G, Leitgeb R A, Javidi B, Anand A 2012 Opt. Lett. 37 5127

    [16]

    Yuan F, Yuan C J, Nie S P, Zhu Z Q, Ma Q Y, Li Y, Zhu W Y, Feng S T 2014 Acta Phys. Sin. 63 104207 (in Chinese) [袁飞, 袁操今, 聂守平, 朱竹青, 马青玉, 李莹, 朱文艳, 冯少彤 2014 物理学报 63 104207]

    [17]

    Singh A S G, Anand A, Leitgeb R A, Javidi B 2012 Opt. Express 20 23617

    [18]

    Lue N, Kang J W, Hillman T R, Dasari R R, Yaqoob Z 2012 Appl. Phys. Lett. 101 084101

    [19]

    Qu W J, Bhattacharya K, Choo C O, Yu Y J, Asundi A 2009 Appl. Opt. 48 2778

    [20]

    Gabai H, Shaked N T 2012 Opt. Express 20 26906

    [21]

    Lan B, Feng G Y, Zhang T, Liang J C, Zhou S H 2017 Acta Phys. Sin. 66 069501 (in Chinese) [兰斌, 冯国英, 张涛, 梁井川, 周寿桓 2017 物理学报 66 069501]

    [22]

    Takeda M, Ina H, Kobayashi S 1982 J. Opt. Soc. Am. 72 156

    [23]

    Cuche E, Marquet P, Depeursinge C 2000 Appl. Opt. 39 4070

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出版历程
  • 收稿日期:  2017-12-22
  • 修回日期:  2018-04-09
  • 刊出日期:  2019-07-20

基于单一分光棱镜干涉仪的双通路定量相位显微术

  • 1. 山东大学信息科学与工程学院, 山东省激光技术与应用重点实验室, 济南 250100;
  • 2. 山东大学物理学院, 济南 250100
  • 通信作者: 卢景琦, Lu618@sdu.edu.cn
    基金项目: 山东省自然科学杰出青年基金(批准号:2013JQE27056)和山东省重点研发计划(批准号:2016GSF121023)资助的课题.

摘要: 仅仅使用一个单独的分光棱镜(BS),实现了一种用于生物细胞三维成像的双通路定量相位显微术.不同于传统的使用方法,将BS倾斜放置,使中央半反射层与入射光光轴之间存在一个非常小的角度.这样基于BS的分光特性,经过BS后的透射光束和反射光束将会叠加在一起并形成干涉.调节样品位置,利用相机拍摄同时获得了存在π相移的双通路干涉图.这种离轴干涉模式,只需要记录单幅干涉图就可以获得真实的相位信息,方法结构简单,易于操作,适用于微小透明样品的三维形貌测量.

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