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

Author:

Michailov Yulia12,Nemerovsky Luba34,Ghetler Yehudith4,Finkelstein Maya35,Schonberger Oshrat67,Wiser Amir34,Raziel Arie35,Saar-Ryss Bozhena12ORCID,Ben-Ami Ido67,Kaplanski Olga34,Miller Netanella34,Haikin Herzberger Einat34ORCID,Mashiach Friedler Yardena34ORCID,Levitas-Djerbi Tali35,Amsalem Eden12,Umanski Natalia12,Tamadaev Valeria12,Ovadia Yaniv S.1ORCID,Peretz Aharon67,Sacks Gilat67ORCID,Dekel Nava67,Zaken Odelya67,Levi Mattan34

Affiliation:

1. Obstetrics and Gynecology Department, Barzilai University Medical Center, Ashkelon 7830604, Israel

2. Faculty of Health Sciences, Ben Gurion University of the Negev, Beer Sheva 8410501, Israel

3. Faculty of Medicine, Tel Aviv University, Tel Aviv 6997801, Israel

4. IVF Unit, Department of Obstetrics and Gynecology, Meir Medical Center, Kfar Saba 4428163, Israel

5. IVF Unit, Wolfson Medical Center, Holon 5822012, Israel

6. IVF Unit, Shaare Zedek Medical Center, Jerusalem 9103102, Israel

7. Faculty of Medicine, Hebrew University, Jerusalem 9112102, Israel

Abstract

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.

Funder

QART Medical

Publisher

MDPI AG

Subject

General Biochemistry, Genetics and Molecular Biology,Medicine (miscellaneous)

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