Dataset for Evaluation of a novel microfluidic chip-like device for purifying bovine frozen-thawed semen for in vitro fertilization

VetCountTM Harvester (MotilityCount ApS, Copenhagen, Denmark) is a novel sperm
purification device. It consists of two chambers separated by a 10 μM microporous
membrane. Untreated semen is applied in one chamber, sperm collection medium
in the other. Motile sperm cells are selected by actively swimming through the
membrane pores into the medium containing chamber. After 30 min incubation,
the sperm collection medium can be aspirated and the purified sperm is ready
for further use.
In a first experiment we assessed sperm quality and recovery of frozen-thawed semen
from six different bulls (n = 6) prior to and after purification with the
VetCountTM Harvester or BoviPureTM gradient centrifugation, a commercial available standard technique. In a second approach, a competitive fertilization assay was performed. Ten straws per bull were pooled, split
in two subsamples, and simultaneously purified either with the VetCountTM Harvester
or BoviPureTM gradient centrifugation. Following purification, sperm cells
from each treatment group were fluorescently labeled with either MitoTrackerTM
Red FM or MitoTrackerTM Green FM. In vitro matured oocytes were inseminated with
equal numbers of red and green labeled sperm. Eighteen hours after fertilization,
fluorescence microscopy was used to determine the origin of the fertilizing spermatozoon.

VetCountTM Harvester（MotilityCount ApS，丹麦哥本哈根）是一款新型精子纯化装置。该装置由被10 μM微孔膜分隔的两个腔室构成：将未处理精液注入其中一个腔室，另一腔室则加入精子收集培养基。活动精子可主动游过膜孔进入装有培养基的腔室，从而实现分选。孵育30分钟后，可抽吸收集精子收集培养基，所得纯化精子即可用于后续实验。
在首个实验中，我们评估了6头不同公牛（n=6）的冻融精液经VetCountTM Harvester或BoviPureTM梯度离心法（一种商用标准技术）纯化前后的精子质量与回收率。第二项实验采用竞争性受精检测方案：将每头公牛的10支精液细管混合并均分为两个亚组，分别使用VetCountTM Harvester与BoviPureTM梯度离心法同步完成纯化。纯化完成后，将各处理组的精子分别用MitoTrackerTM Red FM与MitoTrackerTM Green FM进行荧光标记。将等量的红色与绿色荧光标记精子用于体外成熟卵母细胞的授精。受精18小时后，通过荧光显微镜确定受精精子的来源。