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Table of contents
Chapter I General Introduction I-1 Fluorescence
1 I-1.1 The principle of fluorescence
1 I-1.2 Absorption, excitation and emission spectra
I-1.3 Fluorescence quantum yield, lifetime and brightness
I-1.4 Advantages of fluorescence-based investigative technologies
I-2 Fluorescent reporters
I-2.1 Organic fluorescent probes
I-2.2 Autofluorescent proteins (AFP)
I-2.2.1 Applications of AFPs
I-2.2.2 Limitations of AFPs
I-2.3 Site-specific labeling techniques
I-2.3.1 Biarsenical- tetracysteine tagging system
I-2.3.2 Self-labeling tags: SNAP-tag, CLIP-tag, Halo-tag
I-2.3.3 Fluorogenic site-specific labeling systems
I-2.4 Fluorogen-activating proteins (FAPs)
I-2.5 PYP-tag
I-2.6 Cellular retinoic acid binding protein II (CRABPII)
I-2.7 Spinach
I-2.8 Fluorescence-Activating and absorption-Shifting Tag (FAST)
I-3 Objectives
I-4 References
Chapter II Expansion of the spectral properties of FAST
II-1 Presentation of article 1
II-2 Article 1: Dynamic multi-color protein labeling in living cells
Chapter III Development of far-red emitting FAST
III-1 Molecular engineering of far-red emitting fluorogens
III-1.1 Presentation of article 2
III-1.2 Article 2: Design and characterization of red fluorogenic push-pull chromophores holding great potential for bioimaging and biosensing.
III-2 Directed evolution of protein tags binding and activating far-red-emitting fluorogens
III-2.1 Introduction
III-2.2 Results and discussions
III-2.3 Conclusion and perspective
III-2.4 Materials and Methods
III-2.5 Reference
Chapter IV Development of cell impermeant fluorogens for the selective imaging of cell surface proteins
IV-1 Introduction
IV-2 Results and discussion
IV-3 Conclusion and perspective
IV-4 Materials and Methods
IV-5 Reference
Chapter V General discussion
V-1 Development of new fluorogens
V-2 Selection of new protein tags
