JoVE Logo

Sign In

Keele University

6 ARTICLES PUBLISHED IN JoVE

image

Neuroscience

Flow Cytometry Protocols for Surface and Intracellular Antigen Analyses of Neural Cell Types
Vishal Menon 1, Ria Thomas 1,2, Arun R. Ghale 1,3, Christina Reinhard 1, Jan Pruszak 1,4
1Emmy Noether-Group for Stem Cell Biology, Department of Molecular Embryology, Institute of Anatomy and Cell Biology, University of Freiburg, 2Spemann Graduate School of Biology and Medicine and Faculty of Biology, University of Freiburg, 3School of Life Sciences, Keele University, 4Center for Biological Signaling Studies (BIOSS), University of Freiburg

We provide a detailed description of a protocol for flow cytometric analysis of surface antigens and/or intracellular antigens in neural cell types. Critical aspects of experimental planning, step-by-step methodological procedures, and fundamental principles of flow cytometry are explained in order to enable neurobiologists to exploit this powerful technology.

image

Intra-articular injection of murine MSC for the treatment of inflammation and tissue degradation in Antigen-induced Arthritis.
Alasdair Kay 1, Jim Middleton 2, Oksana Kehoe 3
1Institute for Science and Technology in Medicine, Keele University, 2Faculty of Medicine and Dentistry, University of Bristol, 3Keele University

Here, we present a protocol to induce inflammatory arthritis in vivo using antigen response, termed Antigen-induced Arthritis (AIA), an effective tool for research into Rheumatoid Arthritis (RA) research. Additionally we demonstrate the therapeutic effects of infusion of MSC to reduce inflammation, cartilage degradation and expression of TNFα.

image

Biochemistry

Online Size-exclusion and Ion-exchange Chromatography on a SAXS Beamline
Martha E. Brennich 1, Adam R. Round 2,3, Stephanie Hutin 4
1Structural Biology Group, European Synchrotron Radiation Facility, 2European Molecular Biology Laboratory, 3School of Chemical and Physical Sciences, Keele University, 4Groupe de Microscopie Electronique et Méthodes, Institut de Biologie Structurale

The determination of the solution structure of a protein by small angle X-ray scattering (SAXS) requires monodisperse samples. Here, we present two possibilities to ensure minimal delays between sample preparation and data acquisition: online size-exclusion chromatography (SEC) and online ion-exchange chromatography (IEC).

image

Medicine

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing
Solja Kalha 1, Alison Kuony 1, Frederic Michon 1,2
1Helsinki Institute of Life Science, Institute of Biotechnology, University of Helsinki, 2School of Medicine and Institute for Science and Technology in Medicine, Keele University

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

image

Medicine

Use of a Percutaneous Ventricular Assist Device/Left Atrium to Femoral Artery Bypass System for Cardiogenic Shock
Swethika Sundaravel 1, Poonam Velagapudi 1, Mamas Mamas 2, Sandeep Nathan 3, Alexander Truesdell 4
1University of Nebraska Medical Center, 2Keele Cardiovascular Research Group, Keele University, 3University of Chicago Medicine, 4Virginia Heart/Inova Heart and Vascular Institute

The following article describes the stepwise procedure for placement of a device (e.g., Tandemheart) in cardiogenic shock (CS) that is a percutaneous left ventricular assist device (pLVAD) and a left atrial to femoral artery bypass (LAFAB) system that bypasses and supports the left ventricle (LV) in CS.

image

JoVE Core

Multiplexed Fluorescent Immunohistochemical Staining of Four Endometrial Immune Cell Types in Recurrent Miscarriage
Yiwei Zhao 1, Gene Chi Wai Man 1,2, Loucia Kit Ying Chan 1, Xi Guo 1, Yingyu Liu 3, Tao Zhang 1, Joseph Kwong 1,4, Chi Chiu Wang 1,5, Xiaoyan Chen 3, Tin Chiu Li 1
1Department 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.

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved