基于数字全息显微镜的人类精子的三维动态行为监测技术

doi:10.13418/j.issn.1001-165x.2020.04.021

中国临床解剖学杂志 ›› 2020, Vol. 38 ›› Issue (4): 476-480.doi: 10.13418/j.issn.1001-165x.2020.04.021

基于数字全息显微镜的人类精子的三维动态行为监测技术

孙丽¹，　黄桂²，　龚湘君²，　欧建平¹

1.中山大学附属第三医院生殖医学中心，广州 510630； 2.华南理工大学材料科学与工程学院，广州 510640

收稿日期:2019-08-23 出版日期:2020-07-25 发布日期:2020-07-29
通讯作者: 欧建平，主任医师，硕士生导师，E-mail: oujianping 1968@qq.com
作者简介:孙丽（1981-），女，博士，主治医师，研究方向：生殖内分泌，E-mail：651306521@qq.com

Digital holographic microscopy (DHM) and its application in monitoring three-dimensional dynamics of human spermatozoa

SUN Li¹, HUANG Gui², GONG Xiang-jun² , OU Jian-ping¹

1. Center for Reproductive Medicine, the Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China; 2. Faculty of Material Science and Engineering, South China University of Technology, Guangzhou 510640, P. R. China

Received:2019-08-23 Online:2020-07-25 Published:2020-07-29

摘要/Abstract

摘要： 目的　发展基于数字全息显微镜的人类精子三维动态行为的监测技术。方法　采自健康男性的精子样本分散于人工输卵管液（HTF）中，稀释到2×10⁷/mL后显微制样；利用数字全息显微镜记录包含精子三维位置信息的全息图；平均每组全息图得到背景图；对全息图做背景扣除后，利用定位算法获取精子的实时三维坐标；利用轨迹连接算法将精子的三维坐标连接成三维轨迹；基于三维轨迹，发展算法获取精子的三维运动模式、密度的空间分布、三维速度的空间分布和三维朝向。结果　利用数字全息显微镜，获取了包括精子的三维运动模式及反映精子与表面相互作用的密度分布、三维速度的空间分布和三维朝向在内的多项与精子受精能力相关的信息。结论　数字全息显微镜为分析和评估精子的受精能力提供了更为全面的数据。

关键词: 字全息显微镜, 　人类精子, 　三维动态行为

Abstract: Objective　To apply digital holographic microscopy (DHM) as a tool for monitoring three-dimensional (3D) dynamics of human spermatozoa.　Methods　Sperm samples collected from healthy male participants were diluted to 2×107/mL by artificial human tubal fluid (HTF). These suspensions were injected for further observations. 3D dynamics of human spermatozoa were recorded by DHM with holograms. Background images were obtained by averaging holograms in each subgroup. Background removal was performed and the back-free hologram was applied for 3D localization of the sperm according to Rayleigh-Sommerfeld back propagation algorithm. Instantaneous 3D coordinates of human spermatozoa were linked to form trajectories. Subsequently, 3D motion patterns, density distribution, spatial distribution of 3D velocity and 3D orientation of human spermatozoa were determined.　Results　3D motility parameters, 3D motion patterns as well as information reflecting sperm-surface interactions including density distribution, spatial distribution of 3D velocity and 3D orientation of human spermatozoa were available by utilizing DHM. Conclusions　DHM can provide a comprehensive assessment of sperm fertilizing capacity.

Key words: Digital holographic microscopy, 　Human spermatozoa, 　Three-dimensional dynamics

中图分类号:

R321.1

孙丽, 黄桂, 龚湘君, 欧建平. 基于数字全息显微镜的人类精子的三维动态行为监测技术[J]. 中国临床解剖学杂志, 2020, 38(4): 476-480.

SUN Li, HUANG Gui, GONG Xiang-jun , OU Jian-ping. Digital holographic microscopy (DHM) and its application in monitoring three-dimensional dynamics of human spermatozoa[J]. Chinese Journal of Clinical Anatomy, 2020, 38(4): 476-480.

参考文献 23

[1]	Sigman M. Introduction: male fertility testing: the past, present, and future[J]. Fertil Steril, 2019, 111(5): 833-834.
[2]	翁宝川. 男性不育症患者的精液常规检验结果分析[J]. 基层医学论坛, 2016, 20(7): 949-950.
[3]	Oehninger S, Ombelet W. Limits of current male fertility testing[J]. Fertil Steril, 2019, 111(5): 835-841.
[4]	Vahidi S, Moein M, Nabi A, et al. Effects of microsurgical varicocelectomy on semen analysis and sperm function tests in patients with different grades of varicocele: role of sperm functional tests in evaluation of treatments outcome[J]. Andrologia, 2018, 50(8): e13069.
[5]	Liu DY, Garrett C, Baker HW. Clinical application of sperm-oocyte interaction tests in in vitro fertilization--embryo transfer and intracytoplasmic sperm injection programs[J]. Fertil Steril, 2004, 82(5): 1251-1263.
[6]	胡庭溪, 刘霞, 孙尉峻, 等. 精子动力学分析与其体内、体外受精能力的相关性[J]. 畜牧兽医学报, 2016, 47(8): 1727-1732.
[7]	Nagy Á, Polichronopoulos T, Gáspárdy A, et al. Correlation between bull fertility and sperm cell velocity parameters generated by computer-assisted semen analysis[J]. Acta Vet Hung, 2015, 63(3): 370-381.
[8]	Tomlinson MJ, AsadNaeem. Computer-aided sperm analysis (CASA) in the medical laboratory: CASA in diagnostic andrology and assisted conception[J]. Reprod Fert Develop, 2018, 30(6): 850-859.
[9]	Suarez SS, Pacey AA. Sperm transport in the female reproductive tract[J]. Hum Reprod Update, 2006, 12(1): 23-37.
[10]	Lee SH, Grier DG. Holographic microscopy of holographically trapped three-dimensional structures[J]. Opt Express, 2007, 15(4): 1505-1512.
[11]	Muschol M, Wenders C, Wennemuth G. Four-dimensional analysis by high-speed holographic imaging reveals a chiral memory of sperm flagella[J]. PLoS One, 2018, 13(6): e0199678.
[12]	Angelis AD, Managò S, Ferrara MA, et al. Combined Raman spectroscopy and digital holographic microscopy for sperm cell quality analysis[J]. J Spectrosc, 2017: 9876063.
[13]	Cheong FC, Krishnatrey BJ, Grier DG. Strategies for three-dimensional particle tracking with holographic video microscopy[J]. Opt Express, 2010, 18(13): 13563-13573.
[14]	Qi M, Gong X, Wu B, et al. Landing dynamics of swimming bacteria on a polymeric surface: effect of surface properties[J]. Langmuir, 2017, 33(14): 3525-3533.
[15]	Su TW, Xue L, Ozcan A. High-throughput lensfree 3D tracking of human sperms reveals rare statistics of helical trajectories[J]. PNAS, 2012, 109(40): 16018-16022.
[16]	Sartori P, Chiarello E, Jayaswal G, et al. Wall accumulation of bacteria with different motility patterns[J]. Phys Rev E, 2018, 97(2-1): 022610.
[17]	Kantsler V, Dunkel J, Blayney M, et al. Rheotaxis facilitates upstream navigation of mammalian sperm cells[J]. Elife, 2014, 3: e02403.
[18]	Li G, Tang JX. Accumulation of microswimmers near a surface mediated by collision and rotational Brownian motion[J]. Phys Rev Lett, 2009, 103(7): 078101.
[19]	Berke AP, Turner L, Berg HC, et al. Hydrodynamic attraction of swimming microorganisms by surfaces[J]. Phys Rev Lett, 2008, 101(3): 038102.
[20]	Woolley DM. Motility of spermatozoa at surfaces[J]. Reproduction, 2003, 126(2): 259-270.
[21]	Denissenko P, Kantsler V, Smith DJ, et al. Human spermatozoa migration in microchannels reveals boundary-following navigation[J]. PNAS, 2012, 109(21): 8007-8010.
[22]	Pacey AA, Davies N, Warren MA, et al. Hyperactivation may assist human spermatozoa to detach from intimate association with the endosalpinx[J]. Hum Reprod, 1995, 10(10): 2603-2609.
[23]	Stauss CR, Votta TJ, Suarez SS. Sperm motility hyperactivation facilitates penetration of the hamster zona pellucida[J]. Biol Reprod, 1995, 53(6): 1280-1285.

基于数字全息显微镜的人类精子的三维动态行为监测技术

Digital holographic microscopy (DHM) and its application in monitoring three-dimensional dynamics of human spermatozoa

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