Analysis Of Elemrnts In Single Boar’s Sperm Cell By ICP-MS

Artificial insemination is widely used in pig breeding.The quality of boar semen affects the boar’s reproductive health,production capacity and production profit of the pig breeding industry.Trace elements plays an important role in sperm cell transcription and expression of genetic information,modification and metabolism of biomolecules,transmission and conversion of energy,etc.Therefore,it is necessary to detect the contents of elements in boar’s sperm cells.TRA-ICP-MS can distinguish elemental information between individual cells.This article developed an online dilution injection combined device with a single-channel spray chamber.We used TRA-ICP-MS to analyze the elements of boar’s sperm cells from a single-cell perspective and detected contents of elements in 63 porcine semen samples.The results showed that the online dilution sampling method can improve the efficiency of cell detection.And the sperm cell elements content of healthy boars of different months,different genetic backgrounds and different breeds had little difference.Besides,the contents of P and Mg elements in mouse sperm cells presents different distributions,and the cell heterogeneity is reflected.The main contents are as follows:(1)We designed a single-channel spray chamber which is designed based on Scott double-channel spray chamber of the 7900 ICP-MS.We compared performance of the total injection flow,carrier gas flow and make up gas flow.We also developed an on-line dilution sample injection device combined with a single-channel spray chamber.Using this sampling device,we optimized conditions when testing single yeast cell.It was successfully applied to the element detection of selenium-enriched yeast cells.The results showed that P,Mg,Zn,Se,and Cu present a Gaussian distribution in the selenium-enriched yeast cells,indicating that it is feasible to use an on-line dilution sample injection device to detect elements in the cells.(2)An ICP-MS single-cell analysis method for porcine sperm elements was established.On the basis of laboratory work,the effect of integration time on the signal of pig sperm cells was explored,the conditions of cisplatin incubation of boar’s sperm cell were optimized,and the tail docking experiment was used to analyze the distribution of P,Mg,and Pt elements in sperm cells.The results show that the duration of a single sperm cell is about 0.7ms and the best incubation time for cisplatin is 8h,the best incubation concentration is 10mg/L.In addition,P and Mg are mainly distributed in the head,and Pt is distributed both in the head and tail of sperm.(3)The elemental information of 63 boar’s semen samples including Landrace boars,Yorkshire boars,and Duroc boars were collected by using the online dilution sampling device combined with the single-channel spray chamber.The results showed that the sperm elements content of healthy boars of different breeds,genetic backgrounds and ages had little difference.In addition,the analysis method was applied to the element detection of mouse sperm cells,and the results showed that the P and Mg elements in mouse sperm cells showed different distributions,indicating that it is feasible to use this method to analyze other animal sperm elements.