Introduction to Fluorescence Sensing

Chapter 1. Introduction

Chapter 2. Basic principles

2.1. Overview of strategies in molecular sensing

2.2. Labeled targets in fluorescence assays

2.3. Competitor displacement assays

2.4. Sandwich assays

2.5. Catalytic biosensors

2.6. Direct reagent-independent sensing

Sensing and thinking: How to make the best sensor? Comparison of basic principles

Chapter 3. Theoretical aspects

3.1. Parameters that need to be optimized in every sensor

3.2. Determination of binding constants

3.3. Modeling the ligand binding isotherm

3.4. Kinetics of target binding

3.5. Formats for fluorescence detection

Sensing and thinking: How to provide the quantitative measure of target binding?

Chapter 4. Fluorescence detection techniques

4.1. Fluorescence fundamentals

4.2. Intensity-based sensing

4.3. Anisotropy-based sensing and polarization assays

4.4. Lifetime-based fluorescence response

4.5. Excimer and exciplex formation

4.6. Förster resonance energy transfer (FRET)

4.7. Wavelength-shift sensing

4.8. Two-band wavelength-ratiometric sensing with a single dye

Sensing and thinking: The optimal choice of fluorescence detection technique

Chapter 5. Molecular-size fluorescence emitters

5.1. Fluorophores and their characteristics

5.2. Organic dyes as labels and tags

5.3. Organic dyes as fluorescence reporters

5.4. Visible fluorescent proteins

5.5. Luminescent metal complexes

5.6. Few-atom clusters of noble metals

Sensing and thinking: Which molecular reporter to choose for particular needs?

Chapter 6. Nanoscale fluorescence emitters

6.1. Introduction to light emitting nano-world

6.2. Dye-doped nanoparticles and dendrimers

6.3. Conjugated polymers

6.4. Fluorescent carbon nanostructures

6.5. Semiconductor quantum dots

6.6. Up-converting nanocrystals

Sensing and thinking: Nanoscale emitters, what are the advantages?

Chapter 7. Fluorescent nanocomposites

7.1. Fluorescence enhancement and quenching in nanocomposites

7.2. Modulation of emission parameters in multi-fluorophore systems

7.3. Optical choice of FRET donors and acceptors

7.4. Wavelength referencing, multiplexing and multicolor coding

7.5. Combining fluorescence with magnetic, NMR enhancing and other functionalities

Sensing and thinking: Achieving multitude of functions in designed nanocomposites

Chapter 8. Recognition units

8.1. Multivalency: the principle of molecular recognition

8.2. Recognition units built of small molecules

8.3. Antibodies and their recombinant fragments

8.4. Ligand-binding proteins and protein-based display scaffolds

8.5. Designed and randomly synthesized peptides

8.6. Nucleic acid aptamers

8.7. Peptide nucleic acids

8.8. Molecularly imprinted polymers

Sensing and thinking: Selecting the tools for optimal target recognition

Chapter 9. Mechanisms of signal transduction

9.1. General principles of signal transduction

9.2. Basic signal transduction mechanisms: electron, charge and proton transfer

9.3. Signal transduction via excited-state energy transfer

9.4. Superenhancement and superquenching

9.5. Signal transduction via conformational changes

9.6. Signal transduction via association and aggregation phenomena

9.7. Smart sensing with logical operations

Sensing and thinking: How to couple the recognition and reporting functionalities?

Chapter 10. Supramolecular structures and interfaces for sensing

10.1. Self-assembled supramolecular systems10.2. Building blocks for supramolecular sensors

10.3. Conjugation, labeling and cross-linking.

10.4. Supporting and transducing surfaces.

10.5. Functional lipid and polymer bilayers

Sensing and thinking: Extending sensing possibilities with smart nano-ensembles

Chapter 11. Non-conventional generation and transformation of response

11.1. Chemiluminescence and electrochemiluminescence

11.2. Bioluminescence

11.3. Radioluminescence and Cherenkov effect

11.4. Two-photon excitation and stimulated emission

11.5. Direct optical generation of electrical response signal

11.6. Evanescent-wave fluorescence sensors

11.7. Plasmonic enhancement of luminescence emission

Sensing and thinking: Eliminating light sources and detectors: what remains?

Chapter 12. The sensing devices

12.1. Instrumentation for fluorescence spectroscopy

12.2. Optical waveguides and optodes

12.3. Multi-analyte spotted microarrays

12.4. Suspension arrays and barcoding

12.5. Microfluidic devices.

12.6. Devices incorporating whole living cells

Sensing and thinking: Optimizing convenience, sensitivity and precision for obtaining the proper sensor response

Chapter 13. Focusing on targets

13.1. Temperature, pressure and gas sensing

13.2. Probing the properties of condensed matter

13.3. Detection of small molecules and ions

13.4. Nucleic acid detection and sequence identification

13.5. Recognition of protein targets

13.6. Polysaccharides, glycolipids and glycoproteins

13.7. Detection of harmful microbes

Sensing and thinking: Adaptation of sensor units for multi-scale and hierarchical range of targets

Chapter 14. Sensing inside the living cells

14.1. Modern fluorescence microscopy

14.2. Super-resolution microscopy

14.3. Sensing and imaging on a single molecule level

14.4. Site-specific intracellular labeling and genetic encoding

14.5. Advanced nanosensors inside the cells

14.6. Sensing within the cell membrane

14.7. Sensing different targets in cell interior

Sensing and thinking: Intellectual and technical means for addressing the systems of great complexity

Chapter 15. Sensing the whole body and clinical diagnostics

15.1. Ex-vivo diagnostics

15.2. Sensing the whole body

15.3. Monitoring the cells inside the living body

15.4. Theranostics: combining targeting, imaging and therapy

Sensing and thinking: The strategy of controlling by light of diagnostics and treatment

Chapter 16. Opening new horizons

16.1. Genomics, proteomics and other 'omics'

16.2. The sensors to any target and to immense number of targets

16.3. New level of clinical diagnostics

16.4. Advanced sensors in drug discovery

16.5. Towards sensors that reproduce human senses

16.6. Sensors promising to change the society

Sensing and thinking: Where do we stand and where should we go?

Epilogue.

Appendix. Glossary of terms used in fluorescence sensing

Index