Principles of optics applied to biology.
Applications of physics to the development of novel optical technologies in biotechnology.
Traditional and modern microscopy techniques.
Application of single molecule optical technologies to DNA and molecular motors. Application of microscopy to tissue diagnostics and neuroscience.
J. Mertz – “Introduction to Optical Microscopy” – Roberts & co. publisher
Online tutorials:
- http://www.microscopyu.com/
- http://www.olympusmicro.com/primer
Learning Objectives
Knolewdge acquired:
Principles of optics applied to biology.
Traditional microscopy techniques.
Applications of physics to the development of novel optical technologies in biotechnology.
Modern microscopy techniques.
Novel single molecule techniques. Optical manipulation of a single molecule. Optical tweezers. Mechanics of proteins and nucleic acids. Imaging and localization of single molecules. Probes. Super-resolution microscopy. Application of microscopy to tissue diagnostics. Application of single molecule optical technologies to DNA and molecular motors. Application of microscopy to neuroscience.
Competence acquired
Proper selection of the microscopy technique, tailored to the investigation to be done.
Selection of sample labeling methods for application to different microscopy methods. Optical traps. Image acquisition and quantitative analysis
Skills acquired (at the end of the course):
Use of the microscope, with various imaging methodologies and optical manipulation of biological samples
Prerequisites
Courses recommended: Physics
Teaching Methods
Total hours of the course (including the time spent in attending lectures, seminars, private study, examinations, etc...): 150
Hours reserved to private study and other indivual formative activities: 98
Contact hours for: Lectures (hours): 40
Contact hours for: Laboratory (hours): 12
Further information
Frequency of lectures, practice and lab:
Required
Type of Assessment
Oral exam
Course program
Light and its properties –Electromagnetic radiation – Electromagnetic field – Waves – Maxwell equations – Properties of Electromagnetic waves
Basics of Optics - Wavevector and wavenumber – Propagation in vacuum – Evanescent and radiative field – Reflection and refraction – Lenses – Imaging – Resolution
Molecules and interaction with light –Absorbption – Scattering Rayleigh – Fluorescence – Scattering Raman
Wide-field microscopy – Schematics of a microscope – Illumination – Microscope components – Fluorescence microscopy – Introduction to phase contrast and DIC
Laser scanning confocal microscopy – Laser principles – Optics of laser scanning – Confocal microscope
Biological application of fluorescence - Immuno labeling – Organic dyes – Genetic labeling – Fluorescent protein: GFP
TIRFM – FLIM - FRET – Total Internal Reflection Fluorescence Microscopy –Fluorescence lifetime – Fluorescence Lifetime Imaging Microscopy – FRET (Foster Resonance Energy Transfer)
Two-photon microscopy – Non-linear microscopy – Two-photon excited fluorescence – SHG microscopy – Applications
Vibrational microscopy – Raman micro-spectroscopy – CARS microscopy –SRS microscopy
Optical manipulation – Optical tweezers – Measurements of position and force –Manipulation of cells, sub-cellular structures and single molecules – Mechanics of proteins and nucleic acids – Mechanotransduction.
Imaging and localization of single molecules. Probes Localization of single molecules. Tracking 2D and 3D. MSD.
Super-resolution – Deconvolution microscopy – 4Pi Microscopy – STED microscopy – RESOLFT, PALM and STORM – Examples and applications
Laboratory experiences:
- Clinical application of non-linear microscopy and spectroscopy
- Optical manipulation of single molecules
- Imaging of single-molecule and super-resolution.
- Microscopy applied to neuroscience –