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Basic Science Techniques in Clinical Practice

:	H. R. H. Patel, M. Arya, I. S. Shergill
:	Hitendra R.H. Patel, Manit Arya, Iqbal Shergill
:	Basic Science Techniques in Clinical Practice
:	Springer-Verlag
:	9781846287404
:	1
:	CHF 49.70
:

:	Allgemeines
:	English

:	150
:	Wasserzeichen/DRM
:	PC/MAC/eReader/Tablet
:	PDF

A complete guide to implementing research projects for anyone in the medical professions. This book covers all the main areas, allowing anyone to set up and complete research projects. The techniques outlined here can easily be adapted to clinical projects. Written by international authors to provide a flavor from many institutions, the book's appeal is cross-sectional, both at hospital and primary care levels worldwide. Providing cutting-edge information in an accessible manner, and containing diagrams and easy-to-follow step-by-step guides, this is the first guide of its kind. It contains a complete section on setting up and funding research projects.

	Foreword	5
	Preface	7
	Contents	9
	Contributors	11
	Research Governance	15
	INTRODUCTION	15
	WHY WE NEED RESEARCH GOVERNANCE	16
	HISTORY OF DEVELOPMENT OF RESEARCH GOVERNANCE1,2	16
	SALIENT FEATURES OF THE RESEARCH GOVERNANCE FRAMEWORK	19
	CONCLUSION	20
	References	20
	Designing Health Studies	22
	ESSENTIALS OF A STUDY DESIGN Specifying the Research Question	22
	Selection of Subjects	22
	Specifying the Primary Outcome	22
	Inclusion of a Control Group	23
	Confounding	23
	Sample Size	24
	Bias	24
	Writing a Protocol	25
	TWO TYPES OF STUDIES	25
	Randomized Controlled Trials	25
	Types of Trials	26
	Observational Studies	27
	References	30
	Immunohistochemistry	32
	INTRODUCTION	32
	BASIC IMMUNOHISTOCHEMISTRY	32
	METHODOLOGY	34
	Fixation	35
	Antigen Retrieval	35
	Antibodies	36
	Staining Methods/Detection Systems	37
	Enzyme Labels and Chromogens	40
	Animal Tissue	41
	TROUBLESHOOTING/OPTIMIZATION Optimizing Primary Antibodies	41
	Endogenous Enzyme Activity	42
	Endogenous Biotin	43
	Washing	43
	References	43
	Cell Culturing: A Beginner’s Guide to Understanding the Basics of Cell Culturing	45
	INTRODUCTION	45
	Primary Cells	45
	Cell Lines	45
	FUNDAMENTALS OF CELL CULTURING	46
	What Happens When an Infection Occurs?	46
	Media: Their Function in Cell Culturing	47
	Support Cells	48
	Incubation	49
	CELL EXTRACTION	49
	Handling Tissue Biopsies	49
	Fibroblast Extraction	49
	CONCLUSION	51
	References	51
	Flow Cytometry	52
	WHAT IS FLOW CYTOMETRY?	52
	HISTORICAL ASPECTS OF FLOW CYTOMETRY AND THE FLOW CYTOMETER	53
	FLUORESCENCE	53
	LIGHT SCATTER	55
	COMPONENTS OF THE MODERN FLOW CYTOMETER Fluidic System	55
	Optical System and Analysis	55
	Color Assignment	56
	DATA ANALYSIS Histograms and Dot Plots	57
	Gating and Negative Controls	58
	PRINCIPLES OF FLOW CYTOMETRY SAMPLE PREPARATION	60
	References	60
	Western, Northern, and Southern Blotting	62
	INTRODUCTION: THE HISTORY OF SOUTHERN, NORTHERN, AND WESTERN BLOTTING	62
	BASIC PRINCIPLES AND METHODS: ELECTROPHORESIS, GEL BLOTTING, AND DETECTION	63
	Gel Electrophoresis: Size Separation	63
	Sample Preparation: Southern and Northern	63
	Sample Preparation: Western	63
	Agarose Electrophoresis: Southern and Northern	64
	Sodium Sodecylsulfate–Polyacrylamide Gel Electrophoresis: Western	65
	Blotting: Transfer to Solid Support	66
	Detection and Localization: Complementarity and Hybridization	66
	Blocking	68
	Hybridization	68
	Washing	68
	Detection	68
	Stripping	70
	KEY POINTS	70
	References	70
	Fluorescent In Situ Hybridization	72
	INTRODUCTION	72
	BASIC PRINCIPLES	72
	METHODOLOGICAL ASPECTS OF FISH Probe and Sample Preparation	73
	DNA UNMASKING: PRETREATMENT AND DIGESTION De- waxing and Rehydration	74
	Acid Permeabilization	74
	Reducing Agents	74
	Protease Digestion	74
	Probe and Target DNA Denaturation	75
	Probe and Target DNA Hybridization	75
	POSTHYBRIDIZATION WASH AND VISUALIZATION Posthybridization Wash	76
	Visualization and Scoring/Interpretation of Results	76
	QUALITY ASSURANCE	76
	Slide Pretreatment	76
	Acid Permeabilization	77
	Pretreatment (Reducing Agent)	77
	Protease Digestion	77
	Denaturation	78
	Hybridization	78
	Posthybridization Wash	78
	Visualization and Scoring Slides	79
	References	79
	Quantitative Reverse Transcriptase Polymerase Chain Reaction	80
	INTRODUCTION	80
	UNDERSTANDING THE FUNDAMENTALS	81
	DISCUSSION OF METHODS TO QUANTIFY REAL-TIME POLYMERASE CHAIN REACTION RESULTS	84
	CONTROLS IN QUANTITATIVE REVERSE TRANSCRIPTASE POLYMERASE CHAIN REACTION: USE OF THE HO