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Single Molecule Bio-Physics Single Molecule Fluorescence Techniques - - PowerPoint PPT Presentation
Single Molecule Bio-Physics Single Molecule Fluorescence Techniques - - PowerPoint PPT Presentation
Single Molecule Bio-Physics Single Molecule Fluorescence Techniques Single Molecule Fluorescence Techniques State of the Art imaging of single (immobilized) fluorescent Cy5 molecules Pictures: Tinnefeld Lab Fluorescence Techniques / GFP Green
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Single Molecule Fluorescence Techniques
State of the Art imaging of single (immobilized) fluorescent Cy5 molecules
Pictures: Tinnefeld Lab
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Fluorescence Techniques / GFP Green Fluorescent Protein (GFP) Discovered in Jelly Fish Nobel Prize 2008
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Super-Resolution Microscopy
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Super-Resolution Microscopy
4 s movie of actin labeled Cy5 molecules under 100 µM AA –O2 1 ms integration time Real-time movie Analyzing frame by frame
Pictures: Tinnefeld Lab
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Super-Resolution Microscopy
- J. Vogelsang, T. Cordes, C. Forthmann, C. Steinhauer, and P. Tinnefeld in PNAS 2009, 106, 8107-8112
Pictures: Sauer Lab
Actin Fibers stained with ATTO647
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Fret / Quenching
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Force Spectroscopy
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DNA Force Extension by Magnetic Tweezers
Bustamante Lab
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Applying force to single molecules
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Molecular function of muscle
Pictures: Gaub Lab
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Estimation of entropic forces on a polymer
Pictures: Gaub Lab
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- ATP
+ ATP (2mM)
Pictures: Gaub Lab
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Force Spectroscopy with Optical Tweezers
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Optical Tweezers
A. Ashkin et al., Opt. Lett. 11, 288 (1986) Mie-Regime: Particle >> λ : ray-optics Rayleigh-Regime Particle diameter << lambda Consider particle as electric dipole Typical Trapping wavelength: 1064 nm
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Investigation of Kinesin
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Light Driven Microfluidics
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Lab-on-a-Chip
Controlled Fluid Flow without channels?
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Full Fluid Control
100 µm
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Setup
Fluorescence Microscope x-y-scanner 5 µm IR Laser @ 1450 nm
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What is the driving mechanism?
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Moving warm spot Spot
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Finite Element Analysis
α Expansion Coefficient β
- Temp. Dep. of Viscosity
f Spot Repetition Rate b Spot Width ΔT Spot Temperature
- F. M. Weinert, J. A. Kraus, T. Franosch and D. Braun, Phys. Rev. Lett. 100,
164501 (2008)
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Temperature Imaging
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Dependencies
2
T T T v
f v
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Expansion coefficient and viscosity
v
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More Efficient towards Nanofluidics
2
/ 1 d v
- F. M. Weinert and D. Braun, J. Appl. Phys. 104, 104701 (2008).
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Full Fluid Control
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Microfluidics in Gels
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Pumping in Ice
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Pumping in Ice
- F. M. Weinert, M. Wühr and D. Braun, Appl. Phys. Lett. 94, 113901 (2009)
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Thermophoresis
) exp( T S c c
T
: Thermodiffusion Coefficient : Soret Coefficient
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Towards a Molecule Trap Paternoster
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Towards Accumulation
Thermogravitational Separation Column
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Concentration Problem at the Origin of Life
- P. Baaske, F. M. Weinert, S. Duhr, K. H. Lemke, M. J. Russell and
- D. Braun PNAS 104, 9346 (2007)
Problem for Applications: long equilibration times ~ hours/days
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Linear Clusius Tube
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Temperature Gradient & Bidirectional Flow
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Biderectional Flow
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Thermophoresis + Bidirectional Flow = Accumulation
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Accumulation of 5 base single stranded DNA
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Simulation of 50 base ss DNA
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Vacuum Cleaner for 40nm beads
(real time)
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Vacuum Cleaner for ss 50 base DNA
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40nm bead trap
Polystyrene Spheres D = 40 nm, ST = 0.04 1/K
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Microfluidics in Ice
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