Stain-Free Sperm Analysis and Selection for Intracytoplasmic Sperm Injection Complying with WHO Strict Normal Criteria

Yulia Michailov, Luba Nemerovsky, Yehudith Ghetler, Maya Finkelstein, Oshrat Schonberger, Amir Wiser, Arie Raziel, Bozhena Saar-Ryss, Ido Ben-Ami, Olga Kaplanski, Netanella Miller, Einat Haikin Herzberger, Yardena Mashiach Friedler, Tali Levitas-Djerbi, Eden Amsalem, Natalia Umanski, Valeria Tamadaev, Yaniv S. Ovadia, Aharon Peretz, Gilat SacksNava Dekel, Odelya Zaken, Mattan Levi

Research output: Contribution to journalArticlepeer-review


This multi-center study evaluated a novel microscope system capable of quantitative phase microscopy (QPM) for label-free sperm-cell selection for intracytoplasmic sperm injection (ICSI). Seventy-three patients were enrolled in four in vitro fertilization (IVF) units, where senior embryologists were asked to select 11 apparently normal and 11 overtly abnormal sperm cells, in accordance with current clinical practice, using a micromanipulator and 60× bright field microscopy. Following sperm selection and imaging via QPM, the individual sperm cell was chemically stained per World Health Organization (WHO) 2021 protocols and imaged via bright field microscopy for subsequent manual measurements by embryologists who were blinded to the QPM measurements. A comparison of the two modalities resulted in mean differences of 0.18 µm (CI −0.442–0.808 µm, 95%, STD—0.32 µm) for head length, −0.26 µm (CI −0.86–0.33 µm, 95%, STD—0.29 µm) for head width, 0.17 (CI −0.12–0.478, 95%, STD—0.15) for length–width ratio and 5.7 for acrosome–head area ratio (CI −12.81–24.33, 95%, STD—9.6). The repeatability of the measurements was significantly higher in the QPM modality. Surprisingly, only 19% of the subjectively pre-selected normal cells were found to be normal according to the WHO2021 criteria. The measurements of cells imaged stain-free through QPM were found to be in good agreement with the measurements performed on the reference method of stained cells imaged through bright field microscopy. QPM is non-toxic and non-invasive and can improve the clinical effectiveness of ICSI by choosing sperm cells that meet the strict criteria of the WHO2021.

Original languageEnglish
Article number2614
Issue number10
StatePublished - 23 Sep 2023
Externally publishedYes

Bibliographical note

Publisher Copyright:
© 2023 by the authors.


This study was funded by QART Medical.

FundersFunder number
QART Medical


    • IVF ICSI
    • WHO2021
    • quantitative phase microscopy (QPM)
    • sperm selection


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