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Bioluminescence and Fluorescence for In Vivo Imaging
Author(s): Lubov Brovko
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Book Description

Bioluminescence methods are gaining increased attention due to their sensitivity, selectivity, and simplicity, along with the fact that bioluminescence can be monitored both in vitro and in vivo. This book introduces bioluminescence and fluorescence systems, along with the principles of their application for in vivo imaging of intracellular processes, and covers recent developments in optical (bioluminescence and fluorescence) imaging in cell biology.

This book is intended for scientists and students involved in basic cell physiology research, as well as industry professionals, engineers, and managers involved in drug discovery and pre-clinical drug development. It discusses the practical aspects of luminescence in vivo imaging for monitoring intracellular processes. While some basic knowledge of biochemistry and biophysics is preferable, the book includes a brief review of fundamental principles to allow those not familiar with these disciplines to grasp basic concepts.

Book Details

Date Published: 1 December 2010
Pages: 162
ISBN: 9780819482471
Volume: TT91

Table of Contents
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1 Overview of Bioluminescence; Examples of Bioluminescent Reactions
1.1 History of Bioluminescence
1.2 Bioluminescent System of Bacteria
1.3 Bioluminescent System of Insects (Fireflies)
1.4 Bioluminescent System of Ostracods (Vargula and Cypridina)
1.5 Coelenterazine-based Bioluminescent Systems
    1.5.1 Calcium-dependent photoproteins
    1.5.2 Colenterazine-based enzymatic bioluminescent systems
1.6 Dinoflagellate Bioluminescence
2 Family of Fluorescent Proteins
2.1 Green Fluorescent Protein (GFP) from Aequorea Victoria and Its Mutants: Structure and Spectral Characteristics
2.2 Other Accessory Fluorescent Proteins
2.3 Red Fluorescent Proteins from the Discosoma Genus (DsRed): Structure and Spectral Characteristics
2.4 Various Photoactivatable Fluorescent and Chromoproteins
3 Bioluminescence- and Fluorescence-based Cell Viability and Proliferation Assays
3.1 Principles in Bioluminescent Cell Viability and Proliferation Tests
    3.1.1 Viability/Proliferation tests based on firefly luciferase
    3.1.2 Viability/Proliferation tests based on bacterial luciferase
    3.1.3 Viability/Proliferation tests based on Renilla and Gaussia luciferases
    3.1.4 Fluorescent proteins for cell proliferation and viability assays
3.2 Bioluminescence Environmental Toxicity Tests
3.3 Bioluminescent Viability Tests for Pathogenicity Research and Drug Development
    3.3.1 Investigation of bacteria-host interaction and the efficiency of antimicrobial drugs using in vivo bioluminescence
    3.3.2. Bioluminescent and fluorescent viability tests for oncology research and drug development
3.4 Fluorescence and Bioluminescence Tomography
4 Real-Time In Vivo Monitoring of Gene Expression by Bioluminescence and Fluorescence Imaging
4.1 Luciferase-Based Reporters of Gene Expression
    4.1.1 Bioluminescent gene reporters for clinical research
    4.1.2 Bioluminescent gene expression reporters for physiology research
    4.1.3 Bioluminescence gene expression reporters for viral research and bacteriology
    4.1.4 Bioluminescence gene expression reporters for toxicity testing
4.2 Fluorescent Protein-based Reporters of Gene Expression and Their Applications
5 Bioluminescence and Fluorescence Imaging for In Vivo Real-Time Monitoring of Key Metabolites and Intracellular Environment
5.1 In Vivo Imaging of Intracellular ATP Dynamic
    5.1.1 Bioluminescent in vivo ATP imaging
    5.1.2 Fluorescent in vivo ATP imaging
5.2 Bioluminescent In Vivo Imaging of Calcium Dynamic
    5.2.1 Photoproteins for in vivo calcium imaging
    5.2.2 Fluorescent proteins as intracellular calcium indicators Nonratiometric single fluorescent protein-based GECI Ratiometric GECI based on Fluorescence Resonance Energy Transfer (FRET) between two fluorescent proteins
5.3 In Vivo Optical Imaging of Enzymatic Activities
5.4 In Vivo Fluorescence Imaging of Hydrogen Peroxide
5.5 Genetically-Encoded Fluorescent pH Indicators
5.6 Resonance Energy Transfer-based Sensors for In Vivo Detection of Signal Molecules
    5.6.1 Genetically encoded FRET-based sensor for in vivo cAMP monitoring
    5.6.2 Genetically encoded BRET-based sensor for in vivo cAMP monitoring
6 Bioluminescence and Fluorescence Imaging for In Vivo Monitoring of Protein-Protein Interactions
6.1 Two-hybrid System for In Vivo Monitoring of Protein-Protein Interactions
6.2 Complementation Assay of Protein-Protein Interactions ('Split-Protein' Assay)
6.3 NonRadiative Energy Transfer (FRET/BRET) for Investigation of Protein-Protein Interactions
7 Fluorescence and Bioluminescence Tomography - Current Status and Perspectives
7.1 Fluorescence Molecular Tomography Based on Genetically Engineered Probes
7.2 Bioluminescence Tomography
8 Review of Available Instrumentation for In Vivo Bioluminescence and Fluorescence Imaging
8.1 Plate Readers for Fluorescence and Bioluminescence In Vivo Imaging
8.2 Overview of Systems for Macroscopic/Whole Body Bioluminescence and Fluorescence In Vivo Imaging and Image Processing Software
8.3 Systems for Microscopic Luminescent In Vivo Imaging - Recent Innovations
8.4 Concluding Remarks

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