Flow cytometric and near-infrared Raman spectroscopic investigation of quality in stained, sorted, and frozen-thawed buffalo sperm.
Flow cytometry and Laser Tweezers Raman spectroscopy have been used to investigate Nili-Ravi buffalo (Bubalus bubalis) sperm from different samples (fresh, stained, sorted and frozen-thawed) of the flow-sorting process to optimize sperm sex sorting procedures. During the sorting and freezing-thawing processes, the two detection methods both indicated there were differences in mitochondrial activity and membrane integrity. Moreover, a dispersive-type NIR (Near Infrared Reflection) use of the Raman system resulted in the ability to detect a variety of sperm components, including relative DNA, lipid, carbohydrates and protein contents. The use of the Raman system allowed for PCA (principal components analysis) and DFA (discriminant function analysis) of fresh, stained, sorted and frozen-thawed sperm. The methodology, therefore, allows for distinguishing sperm from different samples (fresh, stained, sorted and frozen-thawed), and demonstrated the great discriminative power of ANN (artificial neural network) classification models for the differentiating sperm from different phases of the flow-sorting process. In conclusion, the damage induced by sperm sorting and freezing-thawing procedures can be quantified, and in the present research it is demonstrated that Raman spectroscopy is a valuable technology for assessing sperm quality.
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, PR China; College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, Henan 471003, PR China.
Xiao-Xia
XX
Li
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, PR China.
Meng
M
Wang
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, PR China.
Huan-Hua
HH
Chen
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, PR China.
Qing-Yang
QY
Li
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, PR China.
Huan
H
Yang
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, PR China.
Hui-Yan
HY
Xu
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, PR China.
Yang-Qing
YQ
Lu
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, PR China.
Ming
M
Zhang
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, PR China. Electronic address: xgyang@gxu.edu.cn.
Xiao-Gan
XG
Yang
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, PR China.
Sheng-Sheng
SS
Lu
State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, Guangxi 530004, PR China. Electronic address: kehuanlu@163.com.
Ke-Huan
KH
Lu
D000818:Animals; D002020:Buffaloes / Q000502:physiology*; D015925:Cryopreservation / Q000662:veterinary*; D004249:DNA Damage; D016002:Discriminant Analysis; D005260:Female; D005434:Flow Cytometry / Q000662:veterinary*; D005615:Freezing; D008297:Male; D016571:Neural Networks, Computer; D025341:Principal Component Analysis; D012662:Semen Preservation / Q000662:veterinary; D012743:Sex Preselection / Q000662:veterinary*; D013059:Spectrum Analysis, Raman / Q000379:methods*; D013194:Staining and Labeling
Buffalo; Flow cytometry; Raman spectroscopy; Sexed sperm