Qingdao Agricultural University 3 articles published in JoVE Immunology and Infection Draining Lymph Node Metastasis Model for Assessing the Dynamics of Antigen-Specific CD8+ T Cells During Tumorigenesis Yan Zhang*1, Xingxing Su*2, Lisha Wang2, Zhengliang Yue2, Qiao Liu3, Ling Ran2, Shun Lei2, Jianjun Hu2, Lifan Xu2, Lilin Ye2, Ping Ji4, Guimei Li1, Qizhao Huang5, Shuqiong Wen4 1College of Veterinary Medicine, Qingdao Agricultural University, 2Institute of Immunology, Third Military Medical University, 3Institute of Cancer, Xinqiao Hospital, Third Military Medical University, 4Stomatological Hospital of Chongqing Medical University, 5Institute of life science, Chongqing Medical University The experimental design presented here provides a useful reproductive model for the studies of antigen-specific CD8+ T cells during lymph node (LN) metastasis, which excludes the perturbation of bystander CD8+ T cells. Biology Staining the Cytoplasmic Ca2+ with Fluo-4/AM in Apple Pulp Lina Qiu1, Daqing Huang1, Yongzhang Wang1, Haiyong Qu1 1College of Horticulture, Qingdao Agricultural University Isolated protoplasts of apple pulp cells were loaded with a calcium fluorescent reagent to detect cytoplasmic Ca2+ concentration. Bioengineering Construction of CRISPR Plasmids and Detection of Knockout Efficiency in Mammalian Cells through a Dual Luciferase Reporter System Hegang Li*1, Huaiyuan Qin*1, Ning Zhang*1, Jinshan Zhao1, Jingjing Xin1, Flor M. Perez-Campo2, Huawei Liu1 1Qingdao Agricultural University, Qingdao, China, 2University of Cantabria, Santander y Torrelavega, Spain Here, we present a protocol describing a streamlined method for the efficient generation of plasmids expressing both the CRISPR enzyme and associated single guide RNA (sgRNAs). Co-transfection of mammalian cells with this sgRNA/CRISPR vector and a dual luciferase reporter vector that examines double-strand break repair allows evaluation of knockout efficiency.