The Chinese University of Hong Kong

20 articles published in JoVE

• Medicine

  
  Application of Optical Coherence Tomography to a Mouse Model of Retinopathy Xiaoting Mai*¹, Shaofen Huang*¹, Wen Chen¹, Tsz Kin Ng^1,2,3, Haoyu Chen^{1 1}Joint Shantou International Eye Center, Shantou University and the Chinese University of Hong Kong, ²Shantou University Medical College, ³Department of Ophthalmology and Visual Sciences, the Chinese University of Hong Kong Here, we describe an in vivo imaging technique using optical coherence tomography to facilitate the diagnosis and quantitative measurement of retinopathy in mice.

• Biology

  
  Multiplexed Fluorescent Immunohistochemical Staining of Four Endometrial Immune Cell Types in Recurrent Miscarriage Yiwei Zhao¹, Gene Chi Wai Man^1,2, Loucia Kit Ying Chan¹, Xi Guo¹, Yingyu Liu³, Tao Zhang¹, Joseph Kwong^1,4, Chi Chiu Wang^1,5, Xiaoyan Chen³, Tin Chiu Li^{1 1}Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, Prince of Wales Hospital, ²Department of Orthopaedics and Traumatology, The Chinese University of Hong Kong, Prince of Wales Hospital, ³Department of Obstetrics and Gynaecology, Shenzhen Baoan Women’s and Children’s Hospital, Shenzhen University, ⁴School of Medicine, Faculty of Medicine and Health Sciences, Keele University, ⁵Li Ka Shing Institute of Health Sciences; School of Biomedical Sciences, The Chinese University of Hong Kong Despite the advancements in multiplex immunohistochemistry and multispectral imaging, characterizing the density and clustering of major immune cells simultaneously in the endometrium remains a challenge. This paper describes a detailed multiplex staining protocol and imaging for the simultaneous localization of four immune cell types in the endometrium.

• Biology

  
  Visualization and Quantification of Brown and Beige Adipose Tissues in Mice using [¹⁸F]FDG Micro-PET/MR Imaging Qing Liu*^1,2, Kel Vin Tan*³, Hing-Chiu Chang³, Pek-Lan Khong³, Xiaoyan Hui^{1,2,4 1}State Key Laboratory of Pharmaceutical Biotechnology, The University of Hong Kong, ²Department of Medicine, Li Ka Shing Faculty of Medicine, The University of Hong Kong, ³Department of Diagnostic Radiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, ⁴School of Biomedical Sciences, Institute of Vascular Medicine, Chinese University of Hong Kong Functional imaging and quantitation of thermogenic adipose depots in mice using a micro-PET/MR imaging-based approach.

• Biochemistry

  
  Density Gradient Ultracentrifugation for Investigating Endocytic Recycling in Mammalian Cells Wai Wa Ray Chan*¹, Yu Qi Zhai*¹, Kwok-Fai Lau^{1 1}School of Life Sciences, The Chinese University of Hong Kong This paper aims to present a protocol for preparing recycling endosomes from mammalian cells using sucrose density gradient ultracentrifugation.

• Behavior

  
  Evaluation of the Cognitive Performance of Hypertensive Patients with Silent Cerebrovascular Lesions Manman Zhang^1,2, Junling Gao³, Bingjiao Xie², Henry Ka Fung Mak⁴, Raymond Tak Fai Cheung^{2,5 1}Department of Neurology, The First Affiliated Hospital of Wenzhou Medical University, ²Department of Medicine, The University of Hong Kong, ³Centre of Buddhist Studies, The University of Hong Kong, ⁴Department of Diagnostic Radiology, The University of Hong Kong, ⁵Research Centre of Heart, Brain, Hormone & Healthy Aging, The University of Hong Kong Here, we present a protocol to assess whether various types of silent cerebrovascular lesions are differentially associated with deficits in certain cognitive domains in a cohort of 398 hypertensive elderly Chinese, using a combination of neuropsychological tests and multi-sequence 3T MRI scanning.

• Developmental Biology

  
  An Enhanced Green Fluorescence Protein-based Assay for Studying Neurite Outgrowth in Primary Neurons Wai Wa Ray Chan*¹, Wen Li*^1,2, Dik Long Dennis Chau*¹, Kwok-Fai Lau^{1 1}School of Life Sciences, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, ²Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen In this report, we describe a simple protocol for studying neurite outgrowth in embryonic rat cortical neurons by co-transfecting with EGFP and the protein of interest.

• Neuroscience

  
  Characterizing the Relationship Between Eye Movement Parameters and Cognitive Functions in Non-demented Parkinson's Disease Patients with Eye Tracking Oscar W.H. Wong¹, Gabriel P.C. Fung², Sandra Chan^{1 1}Department of Psychiatry, The Chinese University of Hong Kong, ²Lab Viso Limited Here, we present a protocol to study the relationship between the eye movement parameters and cognitive functions in non-demented Parkinson's disease patients. The experiment used an eye tracker to measure the saccadic amplitude and fixation duration in a visual search task. The correlation with performance in multi-domain cognitive tasks was subsequently measured.

• Genetics

  
  A Robust Polymerase Chain Reaction-based Assay for Quantifying Cytosine-guanine-guanine Trinucleotide Repeats in Fragile X Mental Retardation-1 Gene Huilin Wang*¹, Xiaofan Zhu*^2,3, Baoheng Gui*^3,4, Wan Chee Cheung², Mengmeng Shi^2,3, Zhenjun Yang^2,3, Ka Yin Kwok², Ricky Lim⁵, Sanna Pietilä⁵, Yuanfang Zhu^6,7, Kwong Wai Choy^{2,3 1}Central Laboratory, Bao'an Maternity and Child Health Hospital, Jinan University, ²Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, ³Shenzhen Research Institute, The Chinese University of Hong Kong, ⁴Department of Genetics and Metabolism, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, ⁵PerkinElmer Diagnostics, ⁶Department of Obstetrics and Gynecology, Bao'an Maternity and Child Health Hospital, Jinan University, ⁷Maternal-Fetal Medicine Institute, Bao'an Maternity and Child Health Hospital, Jinan University An accurate and robust polymerase chain reaction-based assay for quantifying cytosine-guanine-guanine trinucleotide repeats in the Fragile X mental retardation-1 gene facilitates molecular diagnosis and screening of Fragile X syndrome and Fragile X-related disorders with shorter turn-around time and investment in equipment.

• Genetics

  
  Investigation of the Transcriptional Role of a RUNX1 Intronic Silencer by CRISPR/Cas9 Ribonucleoprotein in Acute Myeloid Leukemia Cells Chi-Keung Cheng¹, Terry H.Y. Wong¹, Yuk-Lin Yung¹, Nelson C.N. Chan¹, Margaret H.L. Ng^{1,2 1}Blood Cancer Cytogenetics and Genomics Laboratory, Department of Anatomical and Cellular Pathology, Prince of Wales Hospital, The Chinese University of Hong Kong, ²State Key Laboratory in Oncology in South China, The Chinese University of Hong Kong Direct delivery of preassembled Cas9/guide RNA ribonucleoprotein complexes is a fast and efficient means for genome editing in hematopoietic cells. Here, we utilize this approach to delete a RUNX1 intronic silencer and examine the transcriptional responses in OCI-AML3 leukemic cells.

• Genetics

  
  Semiconductor Sequencing for Preimplantation Genetic Testing for Aneuploidy Baoheng Gui*^1,2,3, Yingxin Zhang*⁴, Bo Liang⁵, Yvonne Ka Yin Kwok⁴, Wai Ting Lui⁴, Queenie Sum Yee Yeung⁴, Lingyin Kong⁶, Liming Xuan⁶, Jacqueline Pui Wah Chung⁴, Kwong Wai Choy^{3,4 1}Department of Genetics and Metabolism, Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region, ²Birth Defects Prevention and Control Institute of Guangxi Zhuang Autonomous Region, ³Shenzhen Research Institute, The Chinese University of Hong Kong, ⁴Department of Obstetrics and Gynaecology, The Chinese University of Hong Kong, ⁵State Key Laboratory of Microbial Metabolism, Joint International Research Laboratory of Metabolic and Developmental Sciences, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, ⁶Basecare Medical Device Co., Ltd Here, we introduce a semiconductor sequencing method for preimplantation genetic testing for aneuploidy (PGT-A) with the advantages of short turnaround time, low cost, and high throughput.

• Cancer Research

  
  Flow Cytometric Detection of Newly-formed Breast Cancer Stem Cell-like Cells After Apoptosis Reversal Yiyue Xu^1,2, Chun So¹, Hon-Ming Lam^1,2, Ming-Chiu Fung^1,2, Suk-Ying Tsang^{1,2,3,4 1}School of Life Sciences, The Chinese University of Hong Kong, ²State Key Laboratory of Agrobiotechnology, The Chinese University of Hong Kong, ³Key Laboratory for Regenerative Medicine, Ministry of Education, The Chinese University of Hong Kong, ⁴Centre for Novel Biomaterials, The Chinese University of Hong Kong Here, we present a protocol to isolate apoptotic breast cancer cells by fluorescence-activated cell sorting and further detect the transition of breast non-stem cancer cells to breast cancer stem cell-like cells after apoptosis reversal by flow cytometry.

• Developmental Biology

  
  A Familial Hypercholesterolemia Human Liver Chimeric Mouse Model Using Induced Pluripotent Stem Cell-derived Hepatocytes Jiayin Yang*^1,2,6, Lai-Yung Wong*², Xiao-Yu Tian*³, Rui Wei^1,2, Wing-Hon Lai², Ka-Wing Au², Zhiwei Luo^4,5, Carl Ward^4,5, Wai-In Ho², David P. Ibañez^4,5, Hao Liu^4,5, Xichen Bao^4,5, Baoming Qin^4,5, Yu Huang³, Miguel A. Esteban^4,5,7, Hung-Fat Tse^{1,2,6,7 1}Department of Medicine, University of Hong Kong-Shenzhen Hospital, ²The Cardiology Division, Department of Medicine, Li Ka Shing Faculty of Medicine, University of Hong Kong, ³School of Biomedical Sciences, Institute of Vascular Medicine, Li Ka Shing Institute of Health Sciences, Chinese University of Hong Kong, ⁴Key Laboratory of Regenerative Biology of the Chinese Academy of Sciences, Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health and Guangzhou Medical University, ⁵Laboratory of RNA, Chromatin, and Human Disease, CAS Key Laboratory of Regenerative Biology and Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, ⁶Research Centre of Heart, Brain, Hormone, and Healthy Ageing, Li Ka Shing Faculty of Medicine, University of Hong Kong, ⁷Hong Kong-Guangdong Stem Cell and Regenerative Medicine Research Centre, University of Hong Kong and Guangzhou Institutes of Biomedicine and Health Here, we present a protocol to generate a human liver chimeric mouse model of familial hypercholesterolemia using human induced pluripotent stem cell-derived hepatocytes. This is a valuable model for testing new therapies for hypercholesterolemia.

• Biology

  
  Magnetic-Activated Cell Sorting Strategies to Isolate and Purify Synovial Fluid-Derived Mesenchymal Stem Cells from a Rabbit Model Zhaofeng Jia^1,2,3, Yujie Liang^4,5, Xingfu Li^2,3, Xiao Xu^1,2,3, Jianyi Xiong^2,3, Daping Wang^1,2,3, Li Duan^{2,3 1}Postgraduate institution, Guangzhou Medical University, ²Guangdong Provincial Research Center for Artificial Intelligence and Digital Orthopedic Technology, ³ This article presents a simple and economic protocol for the straightforward isolation and purification of mesenchymal stem cells from New Zealand white rabbit synovial fluid.

• Engineering

  
  Determination of the Excitation and Coupling Rates Between Light Emitters and Surface Plasmon Polaritons Zhaolong Cao^1,2, Min Lin², Daniel Ong^{2 1}State Key Laboratory of Optoelectronic Materials and Technologies and Guangdong Province Key Laboratory of Display Material and Technology, School of Physics and Engineering, Sun Yat-sen University, ²Department of Physics, The Chinese University of Hong Kong This protocol describes the instrumentation for determining the excitation and coupling rates between light emitters and Bloch-like surface plasmon polaritons arising from periodic arrays.

• Medicine

  
  Detecting Anastasis In Vivo by CaspaseTracker Biosensor Ho Man Tang^1,2, Ming Chiu Fung², Ho Lam Tang^{3 1}Institute for Basic Biomedical Sciences, Johns Hopkins University School of Medicine, ²School of Life Sciences, Chinese University of Hong Kong, ³Department of Neurosurgery, Johns Hopkins University School of Medicine Anastasis is technically challenging to detect in vivo because the cells that have reversed the cell death process can be morphologically indistinguishable from normal healthy cells. Here we describe protocols for detecting and tracking cells that undergo anastasis in live animals by using our newly developed in vivo CaspaseTracker biosensor system.

• Immunology and Infection

  
  A Novel Feeder-free System for Mass Production of Murine Natural Killer Cells In Vitro Patrick Ming-Kuen Tang^1,2, Philip Chiu-Tsun Tang², Jeff Yat-Fai Chung², Jessica Shuk Chun Hung², Qing-Ming Wang², Guang-Yu Lian², Jingyi Sheng², Xiao-Ru Huang², Ka-Fai To¹, Hui-Yao Lan^{2 1}Department of Anatomical and Cellular Pathology, The Chinese University of Hong Kong, ²Li Ka Shing Institute of Health Sciences and Department of Medicine & Therapeutics, The Chinese University of Hong Kong Here, we present a protocol to mass-produce gene-silencing murine NK cells by using a feeder-free differentiation system for mechanistic study in vitro and in vivo.

• Medicine

  
  Using Retinal Imaging to Study Dementia Victor T.T. Chan¹, Tiffany H.K. Tso¹, Fangyao Tang¹, Clement Tham¹, Vincent Mok^2,3,4, Christopher Chen^5,6, Tien Y. Wong^7,8, Carol Y. Cheung^{1 1}Department of Ophthalmology and Visual Sciences, The Chinese University of Hong Kong, ²Department of Medicine & Therapeutics, The Chinese University of Hong Kong, ³Therese Pei Fong Chow Research Centre for Prevention of Dementia, The Chinese University of Hong Kong, ⁴Gerald Choa Neuroscience Centre, The Chinese University of Hong Kong, ⁵Memory Aging and Cognition Centre, National University Health System, ⁶Department of Pharmacology, National University of Singapore, ⁷Singapore Eye Research Institute, Singapore National Eye Centre, ⁸Duke-NUS Medical School, National University of Singapore The retina shares prominent similarities with the brain and thus represents a unique window to study vasculature and neuronal structure in the brain non-invasively. This protocol describes a method to study dementia using retinal imaging techniques. This method can potentially aid in diagnosis and risk assessment of dementia.

• Developmental Biology

  
  A 5-mC Dot Blot Assay Quantifying the DNA Methylation Level of Chondrocyte Dedifferentiation In Vitro Zhaofeng Jia^1,2, Yujie Liang³, Bin Ma^4,5, Xiao Xu^2,6, Jianyi Xiong², Li Duan², Daping Wang^{1,2 1}Guangzhou Medical University, ² We present a method to quantify DNA methylation based on the 5-methylcytosine (5-mC) dot blot. We determined the 5-mC levels during chondrocyte dedifferentiation. This simple technique could be used to quickly determine the chondrocyte phenotype in ACI treatment.

• Biology

  
  In Vivo Biosensor Tracks Non-apoptotic Caspase Activity in Drosophila Ho Lam Tang¹, Ho Man Tang¹, Ming Chiu Fung², J. Marie Hardwick^{1 1}W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins University Bloomberg School of Public Health, ²School of Life Sciences, Chinese University of Hong Kong To detect healthy cells in whole animals that contain low levels of caspase activity, the highly sensitive biosensor designated CaspaseTracker was generated for Drosophila. Caspase-dependent biosensor activity is detected in long-lived healthy cells throughout the internal organs of adult animals reared under optimized conditions in the absence of death stimuli.

• Biology

  
  Strategies for Tracking Anastasis, A Cell Survival Phenomenon that Reverses Apoptosis Ho Lam Tang^1,2,3, Ho Man Tang^1,2,3, J. Marie Hardwick¹, Ming Chiu Fung^{2 1}W. Harry Feinstone Department of Molecular Microbiology and Immunology, Johns Hopkins University Bloomberg School of Public Health, ²School of Life Sciences, Chinese University of Hong Kong, ³Center for Cell Dynamics, Department of Biological Chemistry, Johns Hopkins University School of Medicine The term anastasis refers to the phenomenon in which dying cells reverse a cell suicide process at a late stage, repair themselves, and ultimately survive. Here we demonstrate protocols for detecting and tracking cells that undergo anastasis.