Physics of the living cell
Prerequisiti
Background in biology, chemistry, physics, or mathematics (recommended for third, fourth, fifth year ordinary course students, or postgraduate students)
Programma
Physics of the living cell
(basic principles, open questions, advanced methods)
Introduction:
- From Schrodinger to present: Introduction to Biophysics, applied Science.
Fluorescence microscopy to explore the biological world
- Fluorescence: basic principles and properties
- Methods based on fluorescence:
o Intrinsic cellular fluorophores and label-free microscopy: applications
o Genetically encoded fluorescence: green fluorescent protein and its mutants
o New frontiers: organic fluorophores for live cell imaging
- Fluorescence-based methods to probe molecular interactions:
o Energy transfer by Forster resonance
o Fluorescence anisotropy
o Lifetime of fluorescence and its sensitivity at the nanoscale (measurement and analysis methods)
- Fluorescence-based methods to probe molecular dynamics:
o Perturbation-based methods: e.g. Recovery of fluorescence after photobleaching
o Fluctuation-based methods: from single-point to spatiotemporal (scanning, raster, pair) fluorescence correlation spectroscopy
o Location-based methods: tracking of single molecules
o Orbital tracking and feedback-based imaging
- Fluorescence-based methods to obtain super resolution:
o Microscopy for “chemical expansion”
o Localization methods
o Localization methods using photo-activated fluorophores (PALM and STORM microscopy)
o STED microscopy
Biological case studies:
Case study 1. Membrane heterogeneity: does it exist? What functional role?
Case study 2. Organization of the prokaryotic and eukaryotic cytoplasm: molecular crowding and membraneless organelles
Case study 3. The nuclear pore puzzle: which structure? Which function?
Case study 4. Dynamic measurements in dynamic systems: how to study sub-cellular organelles?
Case study 5. Molecular crowding and its regulation with circadian rhythms: what does the neuron teach us?
Obiettivi formativi
Introduction to the basic principles of cellular biophysics and investigation methods based on optical microscopy at the nanoscale. Presentation of case studies regarding open questions in the field of cellular biophysics at the nanoscale, and discussion of possible strategies to address them
Riferimenti bibliografici
Phillips et al. Physical Biology of the Cell