Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric...

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Single-Molecule Microscopy Group Nonlinear Optics Physical Optics 21-06-2017 Maria Dienerowitz [email protected] www.single-molecule-microscopy.uniklinikum-jena.de

Transcript of Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric...

Page 1: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Single-Molecule Microscopy Group

Nonlinear Optics

Physical Optics

21-06-2017

Maria Dienerowitz

[email protected]

www.single-molecule-microscopy.uniklinikum-jena.de

Page 2: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Introduction

• light matter interactions

• nonlinear optical susceptibility

• phase matching

Nonlinear optics in microscopy

• parametric vs non-parametric

• 2nd harmonic generation (SHG)

• two photon excitation fluorescence (TPEF)

• osteoarthritic cartilage imaging

• 3rd harmonic generation (THG)

• cell lineage reconstruction of early zebrafish embryos

• stimulated Raman scattering (SRS)

• coherent anti-Stokes Raman scattering (CARS)

Contents

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Page 3: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Nonlinear Optics

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nonlinear response

linear response

Hooke’s law

cell division

Page 4: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Classification of optical media

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refractive index

metals n>0 dielectric

n<0 negative-index materials ?

optical frequencies

Page 5: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Speed of light in matter

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speed of light in matter

refractive index

vacuum speed of light

vacuum permeability

vacuum permittivity

relative permittivity relative permeability

Page 6: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Material parameters

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relative permittivity electric susceptibility

permittivity

Page 7: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Constitutive equations

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response of bound charge and current to applied field

displacement field

polarisation density

Page 8: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Nonlinear material response

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polarisation density

electric dipole moment

linear polarisation

2nd order nonlinear polarisation

3rd order nonlinear polarisation

Page 9: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Nonlinear material response

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2nd order nonlinear optical susceptibility

3rd order nonlinear optical susceptibility

atomic fields

laser light

-> 1mW – 1020W

sun light

linear optical susceptibility

Page 10: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Nonlinear material response

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Page 11: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Inversion symmetry

inversion symmetry = centrosymmetric no inversion symmetry = non-centrosymmetric

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Page 12: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Nonlinear material response

inversion-symmetric materials (gases, glasses, fluids, benzene (C6H6),…)

materials with and without inversion symmetry

second-harmonic generation

sum-frequency generation

difference-frequency generation

optical parametric oscillation

third-harmonic generation

self-focussing

saturable absorption

two-photon absorption

stimulated Raman scattering

4-wave mixing (CARS)

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Page 13: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Time-varying polarisation

wave equation in nonlinear optical media

source term

example: second harmonic generation

electric field

2nd order polarisation

optical rectification

2nd harmonic frequency radiation 13

Page 14: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

2nd order interactions

2nd order polarisation

electric field

sum frequency generation (SFG)

difference frequency generation (DFG)

optical rectification (OR)

second harmonic generation (SHG)

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Page 15: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

3rd order interactions

3rd order polarisation

electric field

contains 22 different frequency components (and the negative of each):

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Page 16: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Phase matching

frequency matching condition SHG

SFG

phase matching condition SHG

SFG

temporal phase matching

spatial phase matching

destructive interference

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Page 17: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Phase matching

SHG

same material different frequency

dispersion

normal dispersive material birefringent uniaxial crystal

© A

drian

Pin

gston

e

wavenumber

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Page 18: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Birefringent phase matching

momentum mismatch

coherence length

non critical phase matching temperature matching

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Page 19: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Birefringent phase matching

critical phase matching angle matching

extraordinary rays: -> beam walkoff

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Page 20: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Summary theoretical foundation

source term

frequency matching

phase matching

birefringence

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Page 21: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Introduction

• light matter interactions

• nonlinear optical susceptibility

• phase matching

Nonlinear optics in microscopy

• parametric vs non-parametric

• 2nd harmonic generation (SHG)

• two photon excitation fluorescence (TPEF)

• osteoarthritic cartilage imaging

• 3rd harmonic generation (THG)

• cell lineage reconstruction of early zebrafish embryos

• stimulated Raman scattering (SRS)

• coherent anti-Stokes Raman scattering (CARS)

Contents

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Page 22: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Quantitative cellular imaging

- site specificity

- altering physical or

physiological properties

fluorescent staining

© Justass, Wikipedia

bovine pulmonary artery endothelial cell

green: F-actin (Alexa Fluor 488)

red: mitochondria (MitoTrackerRed)

blue: nuclei (DAPI)

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Page 23: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Nonlinear optics in microscopy

- noninvasive

- intrinsic and often specific chemical contrast

second harmonic generation (SHG) -> non-centrosymmetric structures

microtubule in acute brain slice of 8-month-old wild-type mouse

A Kwan et al., PNAS 105, 11370 (2008), D Debarre et al., Optics Letters 29, 2881 (2004)

third harmonic generation (THG) -> optical heterogeneities (interface)

Drosophila embryo, lipid droplets imaged by THG

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Page 24: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Parametric vs non-parametric

parametric process non-parametric process

-> no net energy transfer -> involves energy loss (or gain)

duration of virtual state

real susceptibility complex susceptibility

absorption complex index of refraction

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Page 25: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Parametric vs non-parametric

parametric process non-parametric process

two photon excitation fluorescence (TPEF)

second harmonic generation (SHG)

sum frequency generation (SFG)

third harmonic generation (THG)

coherent anti-Stokes Raman scattering (CARS)

stimulated Raman scattering (SRS)

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Page 26: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Second harmonic generation - SHG

process, non-centrosymmetric material

2nd harmonic generation

sum frequency generation - SFG

-> collagen

-> microtubules

-> muscle myosin

small excitation volume

decreased background

deeper imaging because of longer λexc

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Page 27: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Two photon excitation fluorescence - TPEF

arrive with 10-18s (1 attosecond) E < 2E1

less photobleaching

1 photon 2 photon

imaging depth: -> confocal laser scanning 50-80μm -> multi-photon imaging 200-1000μm

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Page 28: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

SHG microscopy of early stage osteoarthritic cartilage

R Kumar et al., Biomedical Optics Express 6, 1895 (2015)

characterisation of extra-cellular cartilage matrix -> detection and understanding of progression of Osteoarthritis

SHG – 5μm SHG – 15μm SHG – 30μm TPEF – 30μm

TPEF – 3D SHG – 3D

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Page 29: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Third harmonic generation - THG

interface -> cell surface, intercellular lipid droplets

no molecular asymmetry required

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Page 30: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Cell Lineage Reconstruction of Early Zebrafish Embryos

N Olivier et al., Science 329, 967 (2010)

TPEF + SHG + THG

spherical sample

-> deep imaging: 400μm penetration depth

observe development in real time

-> minute-temporal resolution

high resolution microscopy

-> 1μm spatial resolution

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Page 31: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Structural imaging with THG

N Olivier et al., Science 329, 967 (2010) zebrafish embryo from the one-cell stage

λexc=1200nm

animal pole

vegetal pole

100μm

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Page 32: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Comparison structural detail imaged by THG and TPEF

THG signal TPEF – membrane staining

100μm

N Olivier et al., Science 329, 967 (2010) 32

Page 33: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Nuclear envelope fragmentation revealed by THG

N Olivier et al., Science 329, 967 (2010)

50μm

nuclear envelope dynamics during mitosis

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Page 34: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Mitotic spindle imaging with SHG

N Olivier et al., Science 329, 967 (2010)

SHG mitotic spindle -> microtubules present during mitosis

TPEF H2B/mCherry fusion protein -> chromatin condensation, chromosome formation and displacements

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Page 35: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Mitosis steps revealed by TPEF THG SHG microscopy

N Olivier et al., Science 329, 967 (2010)

SHG mitotic spindle

THG cell contour

TPEF β-actin

overlay

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Page 36: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Raman Scattering – linear optics

non-parameteric process parameteric process

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Page 37: Nonlinear Optics - uni-jena.de · Parametric vs non-parametric parametric process non-parametric process two photon excitation fluorescence (TPEF) second harmonic generation (SHG)

Raman Scattering – nonlinear optics

non-parameteric process

Coherent Anti-Stokes Raman scattering (CARS)

Stimulated Raman scattering (SRS)

parameteric process

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