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Harbin medical university

8 ARTICLES PUBLISHED IN JoVE

Bioengineering

Sample Preparation Strategies for Mass Spectrometry Imaging of 3D Cell Culture Models

Dorothy R. Ahlf Wheatcraft^1,2, Xin Liu^1,2, Amanda B. Hummon^1,2

¹Department of Chemistry and Biochemistry, University of Notre Dame, ²Harper Cancer Research Institute, University of Notre Dame

Immortalized cancer cell lines can be grown as 3D cell cultures, a valuable model for biological research. This protocol describes mass spectrometry imaging of 3D cell cultures, including improvements in the sample preparation platform. The goal of this protocol is to instruct users to prepare 3D cell cultures for mass spectrometry imaging analysis.

Biology

Analysis of Circadian Photoresponses in Drosophila Using Locomotor Activity

Yong Zhang¹, Xueyan Pang¹, Zhenxing Liu¹

¹Department of Biology, University of Nevada Reno

Drosophila locomotor activity is a robust and quantitative measurement of circadian photo-responses. We describe protocols for designing behavior experiments for circadian photo-responses and analyzing the data. Studying the circadian photo-responses is important for dissecting the neuronal and molecular mechanisms of light entrainment.

Developmental Biology

Rapid Acquisition of 3D Images Using High-resolution Episcopic Microscopy

Haochuan Zhang^*1,2,3, JunGang Huang^*2,3,4, Xin Liu^2,3, Ping Zhu⁴, Zhongrong Li¹, Xue Li^2,3

¹Department of Pediatric Surgery, The Second Affiliated Hospital & Yuying Children's Hospital, Wenzhou Medical University, ²Departments of Urology, Boston Children's Hospital, ³Department of Surgery, Harvard Medical School, ⁴Department of Cardiovascular Surgery, Guangdong Cardiovascular Institute, Guangdong General Hospital, Guangdong Academy of Medical Sciences

We describe a detailed protocol using high-resolution episcopic microscopy to acquire three-dimensional (3D) images of mouse embryos. This improved protocol utilizes a modified tissue preparation method to enhance penetration of the fluorescent dye, thereby permitting morphometric analysis of both small and large-sized specimens.

Immunology and Infection

Targeted Antibody Blocking by a Dual-Functional Conjugate of Antigenic Peptide and Fc-III Mimetics (DCAF)

Xue Bai^*1,2, Lin Zhang^*3, Jin Hu², Xiuxiu Zhao², Jinheng Pan², Haiteng Deng³, Shan Feng^1,2

¹Key Laboratory of Structural Biology of Zhejiang Province, School of Life Sciences, Westlake University, ²Mass Spectrometry Core Facility, School of Life Sciences, Westlake University, ³MOE Key Laboratory of Bioinformatics, Center for Synthetic and Systems Biology, School of Life Sciences, Tsinghua University

Development of a dual-functional conjugate of antigenic peptide and Fc-III mimetics (DCAF) is novel for the elimination of harmful antibodies. Here, we describe a detailed protocol for the synthesis of DCAF1 molecule, which can selectively block 4G2 antibody to eliminate antibody dependent enhancement effect during Dengue virus infection.

Education

Optimized Sealing Process and Real-Time Monitoring of Glass-to-Metal Seal Structures

Zhichun Fan¹, Kangjia Hu², Zhiyong Huang¹, Yong Zhang², He Yan¹

¹Institute of Nuclear and New Energy Technology, Key Laboratory of Advanced Reactor Engineering and Safety of Ministry of Education, Collaborative Innovation Center for Advanced Nuclear Energy Technology, Tsinghua University, ²Institute of Nuclear and New Energy Technology, Beijing Key Laboratory of Fine Ceramics, Tsinghua University

Key procedures to optimize the sealing process and achieve real-time monitoring of the metal-to-glass seal (MTGS) structure are described in detail. The embedded fiber Bragg grating (FBG) sensor is designed to achieve online monitoring of temperature and high-level residual stress in the MTGS with simultaneous environmental pressure monitoring.

Biology

Detecting Establishment of Shared Blood Supply in Parabiotic Mice by Caudal Vein Glucose Injection

Xin Liu¹, Xue Bai¹, Mingqi Li¹, Huimin Li¹, Yong Zhang^1,2, Baofeng Yang^1,3

¹Department of Pharmacology, The State-Province Key Laboratories of Biomedicine-Pharmaceutics of China, Key Laboratory of Cardiovascular Research, Ministry of Education, College of Pharmacy, Harbin Medical University, ²Institute of Metabolic Disease, Heilongjiang Academy of Medical Science, ³Department of Pharmacology and Therapeutics, Melbourne School of Biomedical Sciences, Faculty of Medicine, Dentistry and Health Sciences University of Melbourne

Here we describe a new method of detecting successful establishment of shared blood circulation of two parabionts through a caudal vein injection of glucose, which causes minimal damage and is not fatal to the parabionts.

Immunology and Infection

Blocking Lymph Flow by Suturing Afferent Lymphatic Vessels in Mice

Yujia Lin^1,2, Jingna Xue², Shan Liao²

¹Department of General Surgery, The Second Affiliated Hospital of Harbin Medical University, Harbin Medical University, ²Inflammation Research Network, Department of Microbiology, Immunology and Infectious Diseases, Snyder Institute for Chronic Diseases, Cumming School of Medicine, University of Calgary

A protocol to block lymph flow by surgical suturing of afferent lymphatic vessels is presented.

Medicine

Microscopic Replantation of Penile Glans Amputation Due to Circumcision

Da-Chun Jin^*1, Bo Zhou^*1, Jun Li², Cong-Cong Bao¹, Yong Luo¹, Yong Zhang¹, Peng Wang¹, Gang Bi¹, Yan-Feng Li¹

¹Department of Urology, Daping Hospital, Army Medical University, ²Department of Urology, Chongqing 13th People's Hospital

The present protocol describes the emergency management of microscopic replantation of penile glans amputation due to circumcision.