Syllabus
Basic principles of detection of molecular probes - optical (fluorescence, luminiscence, vibrational spectroscopy), spin-based, radioactive, electronmicroscopic, mass-spectrometric
Specific demands of molecular probe applications - in vitro, in fixed cells and tissues, in living cells and tissues, in living organisms; labeling of lipids, nucleic acids, and proteins; instruments for detecting individual molecules and molecular assemblies
Principles of labeling - exogenous (affinity, incl. immunodetection, FlAsH/ReAsH, covalent (click-chemistry, photoaffinity labeling), nucleic acid hybridization (DNA barcoding, DIANA), endogenous (genetically encoded - natural and unnatural amino acids)
Labeling for superresolution optical microscopy - exogenous and genetic labeling, relations to mechanisms of optical superresolution
Fluorescent proteins - spectral and other biophysical properties (structure, reactivity, maturation, photoconversion, immunogenicity, FRET, modification tolerance), non-traditional fluorescent proteins
Self-labeling enzymes - dehalogenases (HALO-tag), methyltrasferases and other alkyltransferases (SNAP-tag, CLIP-tag)
Bioluminiscence - lucipherins and lucipherases, BRET, instrumentation
Raman labels - applications and limitations of linear and non-linear Raman spectroscopy based techniques
Optogenetic approaches - modulation of cell membrane voltage, ion concentration, cellular localization, molecular interactions
Detection of neuronal activity - changes in cell membrane voltage, calcium concentration; detection of activity of neurotransmitters, G-protein signaling and other processes
Labeling in electron, cryoelectron and combined electron/optical microscopy
Annotation
Molecular probes of biological processes.