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South China Normal University

39 ARTICLES PUBLISHED IN JoVE

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Biology

Monitoring Acupuncture Effects on Human Brain by fMRI
Kathleen K. S. Hui 1, Vitaly Napadow 1, Jing Liu 1, Ming Li 1, Ovidiu Marina 1,2, Erika E. Nixon 1, Joshua D. Claunch 1, Lauren LaCount 1, Tara Sporko 1, Kenneth K. Kwong 1
1Department of Radiology, Massachusetts General Hospital and Harvard Medical School, 2William Beaumont Hospital

FMRI and physiological monitoring is used to study the effects of Acupuncture on the central and peripheral nervous systems. Acupuncture mobilizes a limbic-paralimbic-neocortical network, with great overlap with the default mode network, to modulate neurological activity, possibly related to its autonomic effect in the peripheral nervous system.

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Biology

Determination of DNA Methylation of Imprinted Genes in Arabidopsis Endosperm
Matthew Rea 1, Ming Chen 1, Shan Luan 1, Drutdaman Bhangu 1, Max Braud 1, Wenyan Xiao 1
1Department of Biology, Saint Louis University

Imprinting is a phenomenon in plant and mammal reproduction. DNA methylation plays an important role in mechanisms of imprinting. Isolating endosperm and determining methylation status of imprinted genes in Arabidopsis can be difficult. In this protocol, we describe how to isolate endosperm and determine methylation by bisulfite sequencing.

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Medicine

Breathing-controlled Electrical Stimulation (BreEStim) for Management of Neuropathic Pain and Spasticity
Sheng Li 1,2,3
1Department of Physical Medicine and Rehabilitation, University of Texas Health Science Center at Houston , 2UTHealth Motor Recovery Laboratory, TIRR Memorial Hermann Hospital, 3The Institute of Rehabilitation and Research (TIRR), TIRR Memorial Hermann Hospital

The purpose is to present a new method, breathing-control electrical stimulation (BreEStim) for management of neuropathic pain and spasticity.

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Neuroscience

In vivo Interrogation of Central Nervous System Translatome by Polyribosome Fractionation
Wilson Pak-Kin Lou 1, Avni Baser 1, Stefan Klußmann 1, Ana Martin-Villalba 1
1Department of Molecular Neurobiology, German Cancer Research Center (DKFZ)

This protocol illustrates essential modifications of polyribosome fractionation in order to study the translatome of in vivo CNS samples. It allows global assessment of translation and transcription regulation through the isolation and comparison of total RNA to ribosome bound RNA fractions.

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Bioengineering

Micropatterning and Assembly of 3D Microvessels
Meredith A. Roberts *1, Surya S. Kotha *1, Kiet T. Phong 2, Ying Zheng 1,2
1Department of Bioengineering, University of Washington, 2Center for Cardiovascular Biology, Institute for Stem Cell and Regenerative Medicine, University of Washington

This manuscript presents an injection molding method to engineer microvessels that recapitulate physiological properties of endothelium. The microfluidic-based process creates patent 3D vascular networks with tailorable conditions, such as flow, cellular composition, geometry, and biochemical gradients. The fabrication process and examples of potential applications are described.

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Immunology and Infection

SILAC Based Proteomic Characterization of Exosomes from HIV-1 Infected Cells
Collins Cheruiyot 1, Zemplen Pataki 1, Robert Williams 1, Bharat Ramratnam 2,3, Ming Li 3
1Brown University, 2COBRE Center for Cancer Research, Lifespan Laboratories, Rhode Island and Miriam Hospitals, 3Division of Infectious Diseases, Department of Medicine, Warren Alpert Medical School, Brown University

Here, we describe a quantitative proteomics method using the technique of stable isotope labeling by amino acids in cell culture (SILAC) to analyze the effects of HIV-1 infection on host exosomal proteomes. This protocol can be easily adapted to cells under different stress or infection conditions.

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Neuroscience

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method
Qing-Wei Huo *1, Sidra Tabassum *1, Afzal Ali Misrani 1, Cheng Long 1,2
1School of Life Sciences, South China Normal University, 2Brain Science Institute, South China Normal University

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

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Chemistry

In Situ Characterization of Boehmite Particles in Water Using Liquid SEM
Juan Yao 1, Bruce W. Arey 1, Li Yang 1, Fei Zhang 1, Rachel Komorek 1, Jaehun Chun 1, Xiao-Ying Yu 1
1Earth & Biological Sciences Directorate, Pacific Northwest National Laboratory

We present a procedure for real-time imaging and elemental composition analysis of boehmite particles in deionized water by in situ liquid Scanning Electron Microscopy.

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Fabrication of 109° Periodic Domain Walls with a Bottom Electrode in BiFeO3 Thin Films
Deyang Chen *1, Chao Chen *1, Peilian Li 1, Zhen Fan 1, Xingsen Gao 1, Jun-Ming Liu 1,2
1Institute for Advanced Materials, South China Academy of Advanced Optoelectronics, South China Normal University, 2Laboratory of Solid State Microstructures and Innovation Center of Advanced Microstructures, Nanjing University

109° periodic domain walls can successfully be fabricated by introducing a dielectric La-BiFeO3 layer between a bottom electrode and a ferroelectric BiFeO3 layer, enabling the study of the switching behavior of a 109° domain structure and the investigation of the electric field control of exchange bias in a ferromagnet/BiFeO3 system.

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Neuroscience

Whole-cell Currents Induced by Puff Application of GABA in Brain Slices
Yangjian Feng *1,2, Binliang Tang *1,2, Ming Chen 2, Li Yang 1,3,4,5
1School of Psychology, South China Normal University, 2School of Life Sciences, South China Normal University, 3Brain Science Institute, South China Normal University, 4Center for Studies of Psychological Application, South China Normal University, 5Guangdong Key Laboratory of Mental Health and Cognitive Science, South China Normal University

We describe the puff technique, by which pharmacological reagents can be administered during whole-cell patch-clamp recording, and highlight various aspects of the features that are crucial for its success.

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Neuroscience

Concurrent Recording of Co-localized Electroencephalography and Local Field Potential in Rodent
Sungmin Kang 1, Michael Bruyns-Haylett 2, Yurie Hayashi 1, Ying Zheng 1
1School of Biological Sciences, Whiteknights, University of Reading, 2Department of Bioengineering, Imperial College

This protocol describes a simple method for concurrent recording of co-localized electroencephalography (EEG) and multi-laminar local field potential in an anesthetized rat. A burr hole drilled in the skull for the insertion of a microelectrode is shown to produce negligible distortion of the EEG signal.

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Genetics

Embryo Microinjection and Transplantation Technique for Nasonia vitripennis Genome Manipulation
Ming Li 1,2, Michelle Bui 1,2, Omar S. Akbari 1,2
1Department of Entomology and Riverside Center of Disease Vector Research, Institute for Integrative Genome Biology, University of California, Riverside, 2Section of Cell and Developmental Biology, Division of Biological Sciences, University of California, San Diego

Microinjection of Nasonia vitripennis embryos is an essential method for generating heritable genome modifications. Described here is a detailed procedure for microinjection and transplantation of Nasonia vitripennis embryos, which will greatly facilitate future genome manipulation in this organism.

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Bioengineering

Fabricating a Kidney Cortex Extracellular Matrix-Derived Hydrogel
Harrison L. Hiraki 1, Ryan J. Nagao 2, Jonathan Himmelfarb 3, Ying Zheng 2
1Department of Bioengineering, University of Washington, 2Department of Bioengineering, Center for Cardiovascular Biology, Institute for Stem Cell and Regenerative Medicine, University of Washington, 3Department of Medicine, Kidney Research Institute, University of Washington

Here we present a protocol to fabricate a kidney cortex extracellular matrix-derived hydrogel to retain the native kidney extracellular matrix (ECM) structural and biochemical composition. The fabrication process and its applications are described. Finally, a perspective on using this hydrogel to support kidney-specific cellular and tissue regeneration and bioengineering is discussed.

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Neuroscience

Diagnosis of Hirschsprung's Disease by Immunostaining Rectal Suction Biopsies for Calretinin, S100 Protein and Protein Gene Product 9.5
Shuiqing Chi 1, Mijing Fang 1, Kang Li 1, Li Yang 2, Shao-tao Tang 1
1Department of Pediatric Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 2Division of Gastroenterology, Hepatology and Nutrition, Cincinnati Children's Hospital Medical Center

This protocol describes the process of immunostaining rectal suction biopsies for calretinin, S100 protein, and protein gene product 9.5. This novel adjuvant diagnostic method for Hirschsprung's disease has preferable sensitivity and specificity rates.

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Chemistry

Preparation of Poly(pentafluorophenyl acrylate) Functionalized SiO2 Beads for Protein Purification
Sura Kim 1, Jayoung Ku 1,2, Jaemin Park 1, Raisa Kharbash 1,2, Sheng Li 1
1Department of Chemical and Biomolecular Engineering, Korea Advanced Institute of Science and Technology (KAIST), 2KI for Health Science and Technology (KIHST), Korea Advanced Institute of Science and Technology (KAIST)

A protocol for the preparation of poly(pentafluorophenyl acrylate) (poly(PFPA)) grafted silica beads is presented. The poly(PFPA) functionalized surface is then immobilized with antibodies and used successfully for the protein separation through immunoprecipitation.

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Medicine

Analyzing Oxygen Consumption Rate in Primary Cultured Mouse Neonatal Cardiomyocytes Using an Extracellular Flux Analyzer
Shizuko Tachibana 1, Chao Chen 1, Oliver R. Zhang 1, Sarah V. Schurr 1, Cameron Hill 1, Ruixia Li 1, Ana M. Manso 1, Jianlin Zhang 1, Aleksander Andreyev 2, Anne N. Murphy 2, Robert S. Ross 1,3, Yoshitake Cho 1
1Division of Cardiology, Department of Medicine, University of California San Diego, 2Department of Pharmacology, University of California San Diego, 3Cardiology Section, Department of Medicine, Veterans Administration Healthcare, San Diego

The goal of this protocol is to illustrate how to use mouse neonatal cardiomyocytes as a model system to examine how various factors can alter oxygen consumption in the heart.

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JoVE Journal

Experimental Methods for Efficient Solar Hydrogen Production in Microgravity Environment
Katharina Brinkert 1,2, Ömer Akay 3, Matthias H. Richter 1,4, Janine Liedtke 2, Katherine T. Fountaine 5,6, Hans-Joachim Lewerenz 7, Michael Giersig 3,8
1Division of Chemistry and Chemical Engineering, California Institute of Technology, 2European Space Agency/ ESTEC, 3Department of Physics, Freie Universitat Berlin, 4Applied Physics and Sensors, Brandenburg University of Technology Cottbus, 5Resnick Sustainability Institute, California Institute of Technology, 6NG Next, Northrop Grumman Corporation, 7Division of Engineering and Applied Science and Joint Center for Artificial Photosynthesis, California Institute of Technology, 8International Academy of Optoelectronics at Zhaoqing, South China Normal University

Efficient solar-hydrogen production has recently been realized on functionalized semiconductor-electrocatalyst systems in a photoelectrochemical half-cell in microgravity environment at the Bremen Drop Tower. Here, we report the experimental procedures for manufacturing the semiconductor-electrocatalyst device, details of the experimental set-up in the drop capsule and the experimental sequence during free fall.

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Immunology and Infection

Co-staining Blood Vessels and Nerve Fibers in Adipose Tissue
Xin Li 1, Zhengmei Mao 2, Li Yang 1, Kai Sun 1
1Center for Metabolic and Degenerative Diseases, the Brown Foundation of Institute of Molecular Medicine, University of Texas Health Science Center at Houston, 2Microscopy Core, the Brown Foundation of Institute of Molecular Medicine, University of Texas Health Science Center at Houston

New blood vessel formation and sympathetic innervation play pivotal roles in adipose tissue remodeling. However, there remain technical issues in visualizing and quantitatively measuring adipose tissue. Here we present a protocol to successfully label and quantitatively compare the densities of blood vessels and nerve fibers in different adipose tissues.

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Cancer Research

Three-Dimensional Bone Extracellular Matrix Model for Osteosarcoma
Yu Zhang 1, Yupeng Yao 1, Yan Zhang 1
1MOE Key Laboratory of Gene Function and Regulation, School of Life Sciences, Sun Yat-sen University

The bone extracellular matrix (BEM) model for osteosarcoma (OS) is well established and shown here. It can be used as a suitable scaffold for mimicking primary tumor growth in vitro and providing an ideal model for studying the histologic and cytogenic heterogeneity of OS.

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Neuroscience

Evaluation of Hemisphere Lateralization with Bilateral Local Field Potential Recording in Secondary Motor Cortex of Mice
Yunan Chen 1,2, Ming Li 3, Ying Zheng 3, Li Yang 1
1School of Life Sciences, Guangzhou University, 2Institute for Brain Research and Rehabilitation, South China Normal University, 3School of Life Sciences, South China Normal University

We present in vivo electrophysiological recording of the local field potential (LFP) in bilateral secondary motor cortex (M2) of mice, which can be applied to evaluate hemisphere lateralization. The study revealed altered levels of synchronization between the left and right M2 in APP/PS1 mice compared to WT controls.

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Genetics

Direct Gene Knock-out of Axolotl Spinal Cord Neural Stem Cells via Electroporation of CAS9 Protein-gRNA Complexes
Wilson Pak-Kin Lou *1,2, Liqun Wang *3, Cheng Long 1, Lei Liu 3, Ji-Feng Fei 3
1School of Life Sciences, South China Normal University, 2The Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), 3Institute for Brain Research and Rehabilitation (IBRR), South China Normal University

Presented here is a protocol to perform time- and space-restricted gene knock-out in axolotl spinal cords by injecting CAS9-gRNA complex into the spinal cord central canal followed by electroporation.

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JoVE Core

A Reversible Silicon Oil-Induced Ocular Hypertension Model in Mice
Jie Zhang 1,2, Fang Fang 1,3, Liang Li 1, Haoliang Huang 1, Hannah C. Webber 1, Yang Sun 1,4, Vinit B. Mahajan 1,4, Yang Hu 1
1Department of Ophthalmology, Stanford University School of Medicine, 2Department of Ophthalmology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, 3Department of Ophthalmology, Second Xiangya Hospital of Central South University, 4Department of Ophthalmology, Veterans Affairs Palo Alto Health Care

Here, we present a protocol to induce ocular hypertension and glaucomatous neurodegeneration in mouse eyes by intracameral injection of silicone oil and the procedure for silicone oil removal from the anterior chamber to return elevated intraocular pressure to normal.

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Developmental Biology

Isolation, Culture, and Adipogenic Induction of Neural Crest Original Adipose-Derived Stem Cells from Periaortic Adipose Tissue
Yiding Qi *1, Xiang Miao *2, Lian Xu 3, Mengxia Fu 3, Shi Peng 4, Kailei Shi 5, Jun Li 4, Maoqing Ye 5, Ruogu Li 1
1Department of Cardiology, Shanghai Chest Hospital, Shanghai Jiao Tong University, 2Shanghai Institute of Nutrition and Health, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, 3Shanghai Key Laboratory of Hypertension, Shanghai Institute of Hypertension, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, 4Department of Cardiology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 5Department of Cardiology, Shanghai Key Laboratory of Clinical Geriatric Medicine, Huadong Hospital Affiliated to Fudan University

We present a protocol for the isolation, culture, and adipogenic induction of neural crest derived adipose-derived stem cells (NCADSCs) from the periaortic adipose tissue of Wnt-1 Cre+/-;Rosa26RFP/+ mice. The NCADSCs can be an easily accessible source of ADSCs for modeling adipogenesis or lipogenesis in vitro.

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Bioengineering

Cerebral Blood Flow-Based Resting State Functional Connectivity of the Human Brain using Optical Diffuse Correlation Spectroscopy
Chien Poon 1, Ben Rinehart 1, Jun Li 1,2, Ulas Sunar 1
1Department of Biomedical, Industrial and Human Factors Engineering, Wright State University, 2South China Academy of Advanced Optoelectronics, South China Normal University

This protocol demonstrates how to measure resting state functional connectivity in the human prefrontal cortex using a custom-made diffuse correlation spectroscopy instrument. The report also discuss practical aspects of the experiment as well as detailed steps for analyzing the data.

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Genetics

Embryo Microinjection Techniques for Efficient Site-Specific Mutagenesis in Culex quinquefasciatus
Michelle Bui 1, Ming Li 1, Robyn R Raban 1, Nannan Liu 2, Omar S Akbari 1,3
1Section of Cell and Developmental Biology, University of California, San Diego, 2Department of Entomology and Plant Patholog, Auburn University, 3Tata Institute for Genetics and Society, University of California, San Diego

This protocol describes microinjection procedures for Culex quinquefasciatus embryos that are optimized to work with CRISPR/Cas9 gene editing tools. This technique can efficiently generate site-specific, heritable, germline mutations that can be used for building genetic technologies in this understudied disease vector.

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Bioengineering

Dual Raster-Scanning Photoacoustic Small-Animal Imager for Vascular Visualization
Fei Yang 1,2, Zhiyang Wang 1,2, Sihua Yang 1,2
1MOE Key Laboratory of Laser Life Science & Institute of Laser Life Science, College of Biophotonics, South China Normal University, 2Guangzhou Provincial Key Laboratory of Laser Life Science, College of Biophotonics, South China Normal University

A dual raster-scanning photoacoustic imager was designed, which integrated wide-field imaging and real-time imaging.

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Biology

Preparing Irradiated and Marked Male Aedes aegypti Mosquitoes for Release in an Operational Sterile Insect Technique Program
Bianca J. Moreno 1, Robert L. Aldridge 1, Seth C. Britch 1, Barbara E. Bayer 1, Jedidiah Kline 1, Daniel A. Hahn 2, Chao Chen 2, Kenneth J. Linthicum 1
1US Department of Agriculture, Agricultural Research Service Center for Medical, Agricultural, & Veterinary Entomology, 2Department of Entomology and Nematology, University of Florida

The sterile insect technique (SIT) is used to control specific, medically important mosquito populations that may be resistant to chemical controls. Here, we describe a method of mass rearing and preparation of sterile male mosquitoes for release in an operational SIT program targeting the Aedes aegypti mosquito.

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Biology

Identification of Alternative Splicing and Polyadenylation in RNA-seq Data
Gunjan Dixit 1, Ying Zheng 1, Brian Parker 2, Jiayu Wen 1
1Department of Genome Sciences, The John Curtin School of Medical Research, The Australian National University, 2Department of Biology, New York University

Alternative splicing (AS) and alternative polyadenylation (APA) expand the diversity of transcript isoforms and their products. Here, we describe bioinformatic protocols to analyze bulk RNA-seq and 3' end sequencing assays to detect and visualize AS and APA varying across experimental conditions.

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Medicine

Aggravation of Myocardial Ischemia upon Particulate Matter Exposure in Atherosclerosis Animal Model
Yuanmin Yang *1, Shuoqiu Deng *1, Shuiqing Qu 1, Yu Zhang 1, Zhongyuan Zheng 1, Lina Chen 1, Yujie Li 1
1China Academy of Chinese Medical Sciences

This protocol describes a composite animal model with exposure to particulate matter (PM) that aggravates myocardial ischemia with atherosclerosis.

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Developmental Biology

Applications of RNA Interference in American Cockroach
Liang Li *1, Andi Jing *1, Minxin Xie 1, Sheng Li 1,2, Chonghua Ren 1,2
1Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, Institute of Insect Science and Technology & School of Life Sciences, South China Normal University, 2Guangmeiyuan R&D Center, Guangdong Provincial Key Laboratory of Insect Developmental Biology and Applied Technology, South China Normal University

The present protocol describes step-by-step guidelines for the RNAi operation techniques in P. americana.

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Medicine

A Rat Orthotopic Renal Transplantation Model for Renal Allograft Rejection
Hang You 1,2, Xin Mao 2, Chenyang Wang 2, Gang Huang 3, Marcus Groettrup 4,5, Jun Li 2
1School of Medicine, Chongqing University, 2Department of Urological Oncology Surgery, Chongqing University Cancer Hospital, 3Department of Biochemistry and Molecular Biology, College of Basic Medical Science, Army Medical University, 4Division of Immunology, Department of Biology, University of Konstanz, 5Biotechnology Institute Thurgau, University of Konstanz

The rat orthotopic renal transplantation model contributes to investigating the mechanism of renal allograft rejection. The current model increases the recipients' survival without interference with blood supply and venous reflux of the lower body using an end-to-end anastomosis of kidney implantation and an end-to-side "tunnel" method of ureter-bladder anastomosis.

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Medicine

A Modified Cuff Technique for Mouse Cervical Heterotopic Heart Transplantation Model
Xin Mao *1, Peng Xian *1, Hang You 1,2, Gang Huang 3, Jun Li 1
1Department of Urological Oncology Surgery, Chongqing University Cancer Hospital, 2School of Medicine, Chongqing University, 3Department of Biochemistry and Molecular Biology, College of Basic Medical Science, Army Medical University

In the present protocol, a mouse heart transplantation model is used for investigating the mechanism of cardiac allograft rejection. In this heterotopic heart transplantation model, operation efficiency is improved, and the survival of cardiac grafts is ensured by a cervical end-to-end anastomosis of heart implantation using a modified Cuff technique.

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Medicine

Microscopic Replantation of Penile Glans Amputation Due to Circumcision
Da-Chun Jin *1, Bo Zhou *1, Jun Li 2, Cong-Cong Bao 1, Yong Luo 1, Yong Zhang 1, Peng Wang 1, Gang Bi 1, Yan-Feng Li 1
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.

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Medicine

Binocular Dynamic Visual Acuity in Eyeglass-Corrected Myopic Patients
Yuexin Wang *1, Yining Guo *1, Shanshan Wei 2, Yifei Yuan 1, Tingyi Wu 1, Yu Zhang 1, Yueguo Chen 1, Xuemin Li 1
1Department of Ophthalmology, Beijing Key Laboratory of Restoration of Damaged Ocular Nerves, Peking University Third Hospital, 2Beijing Tongren Eye Center, Beijing Tongren Hospital

The present research demonstrates a method to accurately examine dynamic visual acuity (DVA) in myopic subjects with eyeglass correction. Further analysis indicated that the closer the refraction state to emmetropia, the better the eyeglass-corrected binocular DVA is at both 40 and 80 degrees per second.

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Bioengineering

Construction of a Human Aorta Smooth Muscle Cell Organ-On-A-Chip Model for Recapitulating Biomechanical Strain in the Aortic Wall
Mieradilijiang Abudupataer *1, Xiujie Yin *1, Bitao Xiang *1, Nan Chen 1, Shiqiang Yan 2, Shichao Zhu 1, Yang Ming 1, Gang Liu 1, Xiaonan Zhou 1, Hao Lai 1, Chunsheng Wang 1, Kai Zhu 1, Jun Li 1
1Department of Cardiac Surgery and Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, 2Institutes of Biomedical Sciences and the Shanghai Key Laboratory of Medical Epigenetics, Shanghai Medical College, Fudan University

Here, we developed a human aorta smooth muscle cell organ-on-a-chip model to replicate the in vivo biomechanical strain of smooth muscle cells in the human aortic wall.

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Biology

Efficient PAM-Less Base Editing for Zebrafish Modeling of Human Genetic Disease with zSpRY-ABE8e
Shaohui Zheng 1,2, Fang Liang 3, Yu Zhang 1,2, Ji-Feng Fei 4, Wei Qin 1,2, Yanmei Liu 1,2
1Key Laboratory of Brain, Cognition and Education Science, Ministry of Education, 2Institute for Brain Research and Rehabilitation, Guangdong Key Laboratory of Mental Health and Cognitive Science, South China Normal University, 3Institute of Modern Aquaculture Science and Engineering, Guangdong Provincial Engineering Technology Research Center for Environmentally Friendly Aquaculture, Guangdong Provincial Key Laboratory for Healthy and Safe Aquaculture, School of Life Sciences, South China Normal University, 4Department of Pathology, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences

This protocol describes how to perform efficient adenine base editing without PAM limitation to construct a precise zebrafish disease model using zSpRY-ABE8e.

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Neuroscience

Multichannel Extracellular Recording in Freely Moving Mice
Maria Ghouse *1, Meng Li *1, Cheng Long 1,2, Jinxiang Jiang 1
1School of Life Sciences, South China Normal University, 2Panyu Central Hospital Joint Laboratory of Translational Medical Research, Panyu Central Hospital, South China Normal University

The protocol describes the methodology of extracellular recording in the motor cortex (MC) to reveal extracellular electrophysiological properties in freely moving conscious mice, as well as the data analysis of local field potentials (LFPs) and spikes, which is useful for evaluating the network neural activity underlying behaviors of interest.

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Bioengineering

Label-Free Surface-Enhanced Raman Scattering Bioanalysis Based on Au@Carbon Dot Nanoprobes
Ying Zheng 1, Xia Xiao 1, Zhenzhen Li 1, Yong Shao 1, Jieyi Chen 1, Zhouyi Guo 1, Huiqing Zhong 1, Zhiming Liu 1
1MOE Key Laboratory of Laser Life Science & Guangdong Provincial Key Laboratory of Life Science, College of Biophotonics, South China Normal University

In this study, we developed a low-cost surface-enhanced Raman scattering (SERS)-based fingerprint nanoprobe with favorable biocompatibility to show label-free live cell bioimaging and detect two bacterial strains, showing in detail how to get SERS spectra of living cells in a non-destructive method.

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Biology

Isolation and Whole-Cell Patch-Clamp Recording of Hippocampal Microglia from Adult Mice
Yu Chen 1, Cheng Long 1, Li Yang 2
1School of Life Sciences, South China Normal University, 2School of Life Sciences, Guangzhou University

The present protocol describes how to isolate and purify primary hippocampal microglia from adult mice, followed by instructions for conducting whole-cell patch-clamp recordings on these acutely isolated cells.

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