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. 2024 Oct 16;11(10):509. doi: 10.3390/vetsci11100509

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© 2024 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Illustrative flow-cytometric dot plots showing evaluation strategy of analysed fluorescent probes. Firstly, spermatozoa were gated using the FSC/SSC dot plot. DRAQ5 far-red dye was used to distinguish nucleated cells from other events. The specific sperm characteristics were evaluated from the DRAQ5 gated cells. Staining with SYBR-14 and SYTOX Green was used to evaluate viability. The proportion of apoptotic-like cells was evaluated using YO-PRO-1 and Caspase 3/7. Graph showing acrosome-damaged spermatozoa detected using PNA. Evaluation of rabbit sperm mitochondrial activity was using MT Green. Graph showing proportion of capacitated sperm which was detected using FLUO-4 and oxidatively damaged sperm using CellROX Green staining.