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. 2022 Dec 23;13(1):52. doi: 10.3390/ani13010052

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© 2022 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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The proportion of untreated (live) or killed (snap-frozen) sperm positive for PI or LF counted from the image captured with the combined visible/fluorescent image seen in Figure 6. Note that almost all killed sperm were PI+ and LF+ (PI+LF+), but untreated sperm had a lower percentage of this category. On the other hand, untreated sperm had more than 20% PI negative that were also LF− (PI−LF−). Additionally, note that while untreated sperm had about 20% PI+ they were still LF− (PI+LF−), which may suggest that LF binding to sperm is sequential over time; i.e., as sperm deteriorate more, more LF will bind to them. Different letter (A.B.C) indicates significant differences p < 0.001.