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Előadássorozat és PhD kurzus fluoreszcencia-alapú technikákról

2016. május 11.

Prof. Alexander P. Demchenko
(Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv 252030, Ukraine, alexdem@ukr.net)

előadássorozatot tart

Lecture course on fluorescence sensing

címmel Debrecenben, május 30 és június 4 között, közvetlenül az ELMI Kongresszust követően.

Az előadássorozat az ÁOK PhD hallgatói számára PhD kurzusként elszámolható, amelyért kreditpontot kaphatnak.

További információ

Grama László  - laszlo.grama@aok.pte.hu

A kurzus témái

Lecture 1. Introduction to fluorescence.
The origin of light absorption and emission.
Electronic energy levels. Jablonski diagram.
Radiative and non-radiative processes. Quantum yield.
Fluorescence and phosphorescence, delayed fluorescence.
Measurement and presentation of spectra.

Lecture 2. Mechanisms of fluorescence detection.
Intensity-based fluorescence response.
Anisotropy-based response and polarization assays.
Excimers and exciplexes.
Forster resonance energy transfer (FRET)
Display of intermolecular interactions in absorption and emission spectra.
Excited-state electron, charge and proton transfers.
Wavelength ratiometry.

Lecture 3. Basic sensing technologies.
Overview of strategies in molecular sensing.
Labeled targets in fluorescence assays.
Competitor displacement assays.
Sandwich assays.
Direct reagent-independent sensing.
Parameters that need to be optimized in every sensor.
Determination of binding constants.

Lecture 4. Design and properties of reporters: organic dyes, polymers and fluorescent proteins.
The properties of organic dyes, specific requirements for fluorescence reporters.
Trp residue as intrinsic reporter in proteins.
Dye-doped nanoparticles and dendrimers.  
Fluorescent conjugated polymers.
Visible fluorescent proteins.

Lecture 5. Luminescent semiconductor nanocrystals, metal chelating complexes and nanoparticles.
Luminescent chelating complexes of lanthanides.
Phosphorescent metal complexes.
Noble metal nanoparticles and molecular clusters.
Semiconductor Quantum Dots and other nanocrystals.
Up-converting luminophors.
Fluorescent carbon nanoparticles

Lecture 6. Recognition units: from small organic molecules to biopolymers and cells.
Recognition units built from small molecules.
Designed and randomly synthesized peptides and ligand-binding proteins.
Antibodies and their recombinant fragments.
Nucleic acid aptamers. SELEX techniques.
Peptide nucleic acids.
Molecularly imprinted polymers.
Self-assembled macromolecular systems. Affinity coupling.

Lecture 7. Characterization of targets.
Temperature, pressure and gas oxygen sensing.
Characterization of condensed media (including supercritical systems and ionic liquids).
Ion detection (including biologically important calcium, copper and zinc).
Detection of small neutral molecules (e.g. glucose).
Recognition of protein targets.
Nucleic acid detection and sequence identification.
Polysaccharides, glycolipids and glycoproteins.
Detection of harmful microbes.

Lecture 8. Fluorescence instrumentation.
Spectrophotometers and spectrofluorimeters, their optical components.
Lifetime-based instrumentation.
Planar waveguides and surface-sensitive detection.
Spotted microarrays, DNA-chips. Suspension arrays.
Microfluidic devices.
Practical aspects of measurement fluorescence.

Lecture 9. Fluorescence microscopy and single-molecular detection.
Construction of modern fluorescence microscopes.
Evanescent-wave microscopy.
Confocal microscopy.
Two- and multiphoton microscopy.
Time-resolved microscopy.
Super-resolution fluorescence techniques.
Sensing on a single molecule level.

Lecture 10. Future directions in fluorescence sensing.
Genomics, proteomics and other ‘omics’.
The sensors to any target and to immense number of targets.
New level of clinical diagnostics.
Sensing the whole body (Fluorescence tomography).
Advanced sensors in drug discovery.
Sensors promising to change the society.