Research ArticleAPPLIED PHYSICS

High-resolution 4-D acquisition of freely swimming human sperm cells without staining

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Science Advances  10 Apr 2020:
Vol. 6, no. 15, eaay7619
DOI: 10.1126/sciadv.aay7619
  • Fig. 1 Schematic illustrations for sperm cell head orientation recovery algorithm, where the camera plane is parallel to the x-y plane.

    (A) 3-D ellipsoid model, including definition of the pitch, roll, and yaw angles, as well as the three model constants, A, B, and C. (B) The projection of a sperm cell head lying flat. (C) The projection of a sperm cell head lying on its side.

  • Fig. 2 A representative sperm cell head 3-D RI reconstruction from 1000 frames during free swim.

    (A) Middle slices along the three main axes. Color bar denotes RI values. (B) Thresholded 3-D rendering from various perspectives. Two other cells are shown in fig. S5.

  • Fig. 3 3-D dynamic reconstruction of a human sperm flagellum during free swim.

    (A) Digital focusing in action. The white segment moves along the flagellum from the neck to the distal end, in the order indicated by the numbers in the top-left corners, finding the ultimate focus at each location. Color bar indicates phase values in radians. All the images shown are retrieved from a single digital off-axis hologram, acquired in a single camera shot. See dynamic representation in movie S1. (B) 2-D representation of the 3-D segmentation, where the color map indicates the recovered depth (z coordinate). (C) 3-D segmentation result from two viewpoints. See dynamic representation in movie S2.

  • Fig. 4 Full 4-D reconstruction of a sperm cell, obtained from 1000 frames recorded during free swim in 7% PVP.

    (A) Overlay of four frames from the 3-D motion in movie S3, each assigned with a different color. (B) Overlay of 15 frames from the 3-D motion in movie S3, each assigned with a different opacity, where earlier times are more transparent. (C) A single frame from the 3-D motion in movie S3, revealing the internal structure of the sperm cell. Light purple indicates the cell membrane (1.355 ≤ RI < 1.37), green indicates the midpiece (RI = 1.383), yellow indicates the acrosomal vesicle (1.37 ≤ RI < 1.425), red indicated the nucleus (1.425 ≤ RI < 1.465), and dark purple indicates RI ≥ 1.465 (centriole region). (D) Different views of the 3-D head centroid trajectory, changing from blue to red as time progresses.

Supplementary Materials

  • Supplementary Materials

    High-resolution 4-D acquisition of freely swimming human sperm cells without staining

    Gili Dardikman-Yoffe, Simcha K. Mirsky, Itay Barnea, Natan T. Shaked

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    The PDF file includes:

    • Figs. S1 to S9
    • Materials and Methods
    • Legends for movies S1 to S5

    Other Supplementary Material for this manuscript includes the following:

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