Molecular Fluorescent Sensors for Cellular Studies (eBook, PDF)

Redaktion: New, Elizabeth J.

• Format: PDF

Produktbeschreibung

Molecular Fluorescent Sensors for Cellular Studies Enables readers to fully understand the fundamentals and chemical principles of fluorescent sensing and the design of fluorescent sensors Fluorescent sensors are able to provide specific chemical information about cells and can be invaluable in understanding processes that underpin health and disease. Molecular Fluorescent Sensors for Cellular Studies provides an avenue into and overview of currently available fluorescent sensing technology and its application to biological imaging. This book aims to help the reader understand the principles of fluorescence and the mechanisms by which fluorescent sensors operate in order to ensure appropriate and optimal use of sensors. Key applications of fluorescent sensing are presented, with explanations not only of how new sensors can be designed, but also how existing sensors can be applied to various biological settings and conditions. Clear and engaging schematics throughout the book explain chemical principles of sensing to the non-expert. * Discusses the breadth of fluorescent sensors, from commercially available sensors to those reported in literature which are yet to be used widely * Explains how fluorescent sensors operate for appropriate and optimal use from a theoretical standpoint * Provides guidance on how to achieve optimal use of fluorescent sensors in practical settings * Summarizes the principles behind fluorescent sensors and their design This work will be an invaluable resource for postgraduates and professionals in the fields of microscopy, bioimaging, and diagnostic imaging who wish to harness the information to improve practical applications and to gain key knowledge surrounding the many facets of fluorescent sensing. It is also of interest to medical and biological researchers working across industry, universities and medical institutes.

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Produktdetails

• Verlag: John Wiley & Sons
• Seitenzahl: 304
• Erscheinungstermin: 15. August 2022
• Englisch
• ISBN-13: 9781119749806
• Artikelnr.: 65464203

Autorenporträt

Elizabeth J. New undertook her undergraduate and Masters studies at the University of Sydney before completing her PhD studies in 2010 at the University of Durham (UK), followed by postdoctoral training at UC Berkeley. Her research focuses on the development of small molecule fluorescent sensors for biological and environmental applications. Her novel sensors for oxidative stress, hypoxia, and essential and therapeutic metal ions are being widely used in research studies around the world.

Inhaltsangabe

List of Contributors ix

1 An Introduction to Small Molecule Fluorescent Sensors 1
Liam D. Adair, Kylie Yang, and Elizabeth J. New

1.1 What is Fluorescence? 1

1.2 Why Is Fluorescence Useful? 3

1.3 What Is a Fluorescent Sensor? 4

1.4 General Types of Fluorescent Sensors 5

1.5 Important Parameters 7

1.5.1 Excitation Maxima 7

1.5.2 Emission Maxima 8

1.5.3 Stokes Shift 8

1.5.4 Quantum Yield 9

1.5.5 Molar Extinction Coefficient 9

1.5.6 Brightness 10

1.5.7 Lifetime 10

1.5.8 Photobleaching 11

1.5.9 Anisotropy 12

1.5.10 Quenching 12

1.6 Fluorescence Mechanisms Used in Fluorescent Sensors 13

1.6.1 Photoinduced Electron Transfer 13

1.6.2 Internal Charge Transfer 14

1.6.3 Förster Resonance Energy Transfer 15

1.6.4 Through Bond Energy Transfer 17

1.6.5 Excited- State Intramolecular Proton Transfer 18

1.6.6 Aggregation- Induced Emission 19

1.6.7 Excimer Formation 21

1.7 Commonly Used Fluorophores 21

1.7.1 Fluorescein 22

1.7.2 Rhodamine 24

1.7.3 Coumarin 26

1.7.4 Naphthalimide 26

1.7.5 BODIPY (4,4- Difluoro4- bora- 3a,4a- diaza- s- indacene) 27

1.7.6 Cyanine 28

1.8 Summary 30

References 30

2 The Applications of Responsive Fluorescent Sensors to Biological Systems 37
Jia Hao Yeo and Elizabeth J. New

2.1 Criteria for Biologically Relevant Fluorescent Sensors 37

2.2 Microscopy for Visualising Fluorescent Sensors 39

2.2.1 Important Considerations in Microscopy 39

2.2.1.1 Resolution in Microscopy 39

2.2.1.2 Understanding the Competition Between True Signal and Noise 41

2.2.1.3 Phototoxicity in Cells 42

2.2.2 Common Microscopy Techniques 43

2.2.2.1 Fluorescence Microscopy 43

2.2.2.2 Confocal Microscopy 44

2.2.2.3 Multiphoton Microscopy 45

2.2.2.4 Fluorescence Lifetime Imaging Microscopy 45

2.2.2.5 Other Advanced Microscopy Techniques 47

2.3 Other Instrumental Techniques for Studying Cells Treated with Fluorescent Sensors 49

2.3.1 Flow Cytometry 49

2.3.1.1 Principles of Flow Cytometry 49

2.3.1.2 Understanding Flow Cytometry Data with Small- molecule Sensors 50

2.3.1.3 Recent Advances in Flow Cytometry 51

2.3.2 Fluorescence Plate- readers 51

2.3.2.1 Standard Plate- reader Assays 51

2.3.2.2 High- content Imaging (HCI) Plate- readers 52

2.4 Biological Samples to Which Fluorescent Sensors Can Be Applied 52

2.4.1 Cultured Mammalian Cells 52

2.4.1.1 Adherent Mammalian Cells 53

2.4.1.2 Non- adherent Cells 53

2.4.1.3 Multi- cellular Models 54

2.4.2 Bacteria 54

2.4.3 Plants 54

2.4.4 Multi- cellular Organisms 55

2.4.5 Towards In Vivo Imaging 55

2.5 Common Challenges and Misconceptions in the Applications of Fluorescent Sensors 56

2.5.1 Important Considerations in Applying Sensors 56

2.5.2 Common Misconceptions About the Use of Sensors - The Bridge Between Multiple Disciplines 57

2.6 Conclusions 60

References 60

3 Methods to Control the Subcellular Localisation of Fluorescent Sensors 63
Jiarun Lin, Kylie Yang, and Elizabeth J. New

3.1 Introduction 63

3.2 Targeting the Nucleus 64

3.3 Targeting Mitochondria 66

3.4 Targeting Lysosomes 67

3.5 Targeting Endosomes 69

3.6 Targeting Autophagic Compartm