EMERGING TECHNOLOGIES

Basic Microscopy: Light Microscopy: lenses and microscopes, resolution: Rayleigh’s Approach, Darkfield; Phase Contrast; Differential Interference Contrast; fluorescence and fluorescence microscopy: what is fluorescence, what makes a molecule fluorescent, fluorescence microscope; optical arrangement, light source; filter sets: excitation filter, dichroic mirror, and barrier, optical layout for image capture; CCD cameras; back illumination, binning; recording color; three CCD elements with dichroic beamsplitters, boosting the signal.

Advanced Microscopy: Confocal microscope: scanning optical microscope, confocal principle, resolution and point spread function, light source: gas lasers & solid-state, primary beamsplitter; beam scanning, pinhole and signal channel configurations, detectors; pixels and voxels; contrast, spatial sampling: temporal sampling: signal-to- noise ratio, multichannel images. nonlinear microscopy: multiphoton microscopy; principles of two-photon fluorescence, advantages of two-photon excitation, tandem scanning (spinning disk) microscopes, deconvolving confocal images; image processing, three-dimensional reconstruction; advanced fluorescence techniques: FLIM, FRET, and FCS, Fluorescence Lifetime, Fluorescence Resonant Energy Transfer (FRET), Fluorescence Correlation Spectroscopy (FCS), Evanescent Wave Microscopy; Near-Field and Evanescent Waves, Total Internal Reflection Microscopy; Near-Field Microscopy; Beyond the Diffraction Limit: Stimulated Emission Depletion (STED), Super-Resolution Summary, Super-Resolution Imaging with Stochastic Optical Reconstruction Microscopy (STORM) and Photoactivated Localization Microscopy (PALM).

Ionization techniques; mass analyzers/overview MS; FT-ICR and Orbitrap, fragmentation of peptides; proteomics, nano LC-MS; Phospho proteomics; interaction proteomics, mass spectroscopy in structural biology; imaging mass spectrometry. High throughput screens in cellular systems, target identification, validation of experimental methods to generate the omics data, bioinformatics analyses, mathematical modeling and designing testable predictions.

X-ray diffraction methods, solution & solid-state NMR, cryo-electron microscopy, small-

angle X-ray scattering, Atomic force microscopy. History of its discovery, elucidation of the mechanism including introduction to all the molecular players, development of applications for in vivo genome engineering for genetic studies, promise of the technology as a next generation therapeutic method.

Introduction to nanobodies, combining nanobody with phage-display method for development of antibody against native proteins, nanobody as a tool for protein structure-function studies, use of nanobodies for molecular imaging, catabolic antibodies using nanobodies.