Chinese Journal of Clinical Anatomy ›› 2020, Vol. 38 ›› Issue (4): 476-480.doi: 10.13418/j.issn.1001-165x.2020.04.021

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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. 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

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

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