Table of Contents

List of contributors ix Foreword xi Preface xiii Acknowledgements xvii Introduction xix Section A - Basic principles 1 1 Philosophy of science 3 James Ladyman 2 Ingredients of experimental design 9 Nick Colegrave 3 Statistics: a journey that needs a guide 17 Gordon Drummond Section B - Cell and molecular 27 4 Organ bath pharmacology 29 Emma Robinson 5 Small vessel myography 39 Tomoko Kamishima and John M Quayle 6 Mammalian cell cultures: the example of airway epithelial cell cultures for cystic fibrosis research 49 Scott H Randell 7 Electron microscopy (TEM and SEM) 59 Paul Verkade 8 Fluorescence microscopy 67 Mark Jepson 9 Intracellular 'sharp' microelectrode recording 77 Helena C Parkington and Harold A Coleman 10 Single electrode voltage-clamp (SEVC) 85 Harold A Coleman and Helena C Parkington 11 Patch clamp recording 95 Neil Bannister and Phil Langton 12 Production of antibodies 105 Elek Molnar 13 Immunocytochemistry and immunohistochemistry 117 Elek Molnar 14 Immunoprecipitation (IP) 129 David Bates 15 Immunoblotting (western) 137 Samantha F. Moore, Joshua S. Savage and Ingeborg Hers 16 Applications of green fluorescent protein (GFP) 147 Mark Jepson 17 Fluorescent measurement of ion activity in cells 153 Helen Kennedy 18 Detection of exocytosis -- real time 163 Anja Teschemacher 19 Viral vector transgenesis 173 Anja Teschemacher 20 Polymerase chain reaction (PCR) and reverse transcription (RT)-PCR 179 Lucy F. Donaldson 21 In situ hybridisation (ISH) 187 Lucy F. Donaldson 22 Methods of interference (antisense, siRNAs and dominant negative mutations) 193 Allison Fulford 23 Transcriptome analysis: microarrays 203 Charles Hindmarch 24 Experimental proteomics 215 Thierry Le Bihan Section C - In vivo / integrative 229 25 Behavioural methodology 231 Emma Robinson 26 Genetically modified mouse models 241 Nina Balthasar 27 Wireless recording of cardiovascular signals 247 Julian FR Paton and Fiona D McBryde 28 Electrical stimulation methods 253 Jon Wakerley 29 Extracellular recording 261 Jon Wakerley 30 Antidromic identification 271 Jon Wakerley 31 Event-triggered averaging, including spike-triggered averaging 279 Richard Apps 32 Axonal transport tracing of CNS pathways 285 John Crabtree 33 Cardiovascular methods: general considerations for human studies 291 Erica A Wehrwein and Michael J Joyner 34 Measuring cardiac output in humans 299 Erica A Wehrwein and Michael J Joyner 35 Measuring peripheral blood flow in humans 311 Erica A Wehrwein and Michael J Joyner Index 319

About the Author

Dr. Philip David Langton, Senior Lecturer and Academic Director of E-Learning, Department of Physiology, University of Bristol
Dr Philip Langton is currently a Senior Lecturer and Academic Director of e-Learning in the Faculty of Medical and Veterinary Sciences at the University of Bristol, UK.?Dr Langton is an advocate for excellence in all forms of learning, teaching and assessment in HE. He is an enthusiastic undergraduate and graduate teacher, contributing to Medical Sciences and Physiological Sciences programmes. Finalist of 'Bioscience Teacher of the Year' for 2011, Dr Langton is passionate about providing an excellent learning experience for students in ways that are stimulating and rewarding for University staff, and disseminating innovative and high-quality learning practices.

Reviews

"Too many recent bioscience graduates lack significant research lab experience. Even research projects can expose them to just a few techniques. Appraisal of research papers in undergraduate courses tends to address the results, but rarely includes a critical evaluation of the researchers methodology. In reality, few bioscientists can claim a working knowledge of more than a handful of the techniques covered in this collection. These shortcomings can be greatly overcome thanks to this book. The contributors are active research scientists of high quality. Each addresses the methods in a critical sense and provide an expert's view of the advantages and pitfalls. There is no equivalent book currently available." Dr David J. Miller on behalf of The Physiological Society "Research questions require the scientist to employ molecular, cell, organism and population approaches, and the reader of scientific literature to carefully consume data from each of those environments. This book provides a clear and concise summary of the research techniques and approaches that generate data. Importantly, the authors identify appropriate uses and cautionary caveats, essential for readers who do not use the technique. This book is essential for accurate and careful interpretation of scientific literature." Professor Robert G. Carroll, Editor Advances in Physiology Education