1. Looking for New Instruments for Scientific Research
전승준

2. 과학발전에서 연구 장비의 중요성 X-ray, NMR, Mass, etc – Nobel winners
For big breakthrough in science,
we needed new instruments
- 근대과학혁명 : 망원경 (갈릴레오, 뉴튼)
20세기 양자물리학 완성 : X-ray, 분광기, 음극관 등
Bio era : Diffraction, Mass, NMR, Microscopy ….
결국 공정장비, 의료장비 등 산업장비에 응용

3. Instruments for Scientific Research
분석(Analysis) : separation, property (분석화학)
structure, dynamics (물리화학)
- Chromatography
- Thermodynamics : calorimeter
- Mass
- Hydrodynamics : sedimentation
- Optical Spectroscopy
- Microscopy : single molecular detection
- Diffraction :X-ray, neutron, electron
NMR
제조(Synthesis, Processing)

4. Journey of My Quest in Science
Ion beam 실험(Mass)
Radical의 고압 안정화 연구
(DAC, Spectroscopy)
고체수소 및 산소 고압연구
분광학과 고압, 계산화학
비선형 광학물질연구 &
선형/비선형 형광 현미경
(Femto-Laser, Microscopy)
06-현재 다차원/비선형 분광학

5. 1980년대 초반 연구 : Mass - Radical 연구

6. 1980년대 후반 연구 : 고압 + 분광학 연구
Diamond Anvil Cell
Spectroscopy

7. 1990년대 초 중반 화학 분야 관심사 Materials! Nano?-X
Technology Vision 2020 : The US Chemical Industry
(American Chemical Society,American Institute of Chemical Engineers
Chemical Manufacturers Association, Council for Chemical Research,
Synthetic Organic Chemical Manufacturers Association)
Chemical Science
1. Chemical Synthesis
2. Bioprocesses and Biotechnology
3. Materials Technology
“What Will Chemistry Do in the Next Twenty Years?”
(G. Whitesides , Harvard 대 화학과 ,Angewante1991 )
1. Materials Chemistry
2. Biological Chemistry
3. Computational Chemistry
4. Small Basic Science
Materials! Nano?-X

8. Nonlinear Optical Phenomena
Interaction between Matter and Light
Molecular View : dipole moment m
: linear (first-order) polarizability
: second-order polarizability (first hyperpolarizability)
: third-order polarizability (second hyperpolarizability)
Macroscopic View : Polarization
: Linear (first-order)susceptibility
: Second-order susceptibility
: Third-order susceptibility
: rank i + 1 tensor

9. What is Nonlinear Optical (NLO) Materials ?
The materials which exhibit a large non-linear dependence of
optical phenomena on the light intensity
Nonlinear Optical Phenomena
Application
-Second Harmonic Generation
(Frequency doubling)
-Pockels Effect
-Third Harmonic Generation
(Frequency Tripling)
- Kerr Effect
- Degenerate Four-Wave mixing
-Frequency doubling, tripling, mixing
-Electro-Optic Modulator
-Optical Switching
-Optical Information Storage

10. Mach-Zehnder Modulator

11. Linear Push-Pull Polyenes
(Dipolar Molecules)

12. Materials Polarization
c(2) = 0
c(2) >> 0 E

13. Octupolar Molecules

14. Octupole
Dipole

15. Dipole vs Octupole Dipolar Molecules Octupolar Molecules
Push-Pull Polyenes:
Dipole-dipole interaction favors anti-parallel alignment.
Difficult to arrange dipolar molecules to maximize NLO properties.
Octupolar Molecules
Nonpolar molecules with threefold rotational symmetry:
No need to overcome dipole-dipole interaction.
Easy to arrange molecules in the same direction in the solid state.

16. VB-3CT Model Hamiltonian for Crystal Violet

17. Design Strategy for the Octupolar NLO Molecules
1. Strong donor-acceptor pair
2. Increased p-orbital energy
3. Extended conjugation
  2,   3,   4 ( = qd)

18. Octupolar Supramolecules

19. Synthesis of 1,3,5-Trinitro-2,4,6-tris(styryl)benzene Derivatives
J. Am. Chem. Soc. 2001, 123,

20. Synthesis of 1,3,5-Tricyano-2,4,6-tris(styryl)benzene Derivatives

21. Absorption & Emission Spectrum of
1,3,5-Tricyano-2,4,6- tris(p-diethylaminostyryl)benzene

22. Experimental Set-up for HRS
OPO L2
PBP P1 BS
Q-switched
Nd:YAG Laser
1064 nm
Sample Cell
1560 nm PD L3 P2 CL L4 BF PC
Boxcar
PMT

23. Hyper-Rayleigh scattering signal for
1,3,5-tricyano-2,4,6-tris(p-diethylaminostyryl)benzene
S.-J. Jeon et al.
Chem. Phys. Lett., 261, 307 (1996)
N.M.Song, T-I.Kang. S. C. Jeoung. S.-J. Jeon, B. T. Cho, D. Kim,
Chem. Phys. Lett., 261, 307 (1996)
N.M.Song, T-I.Kang. S. C. Jeoung. S.-J. Jeon, B. T. Cho, D. Kim,
Chem. Phys. Lett., 261, 307 (1996)
N.M.Song, T-I.Kang. S. C. Jeoung. S.-J. Jeon, B. T. Cho, D. Kim,
Chem. Phys. Lett., 261, 307 (1996)

24. Normalized Internal Reference Method
1,3,5-Tris(p-piperidylstyryl)-2,4,6-trinitrobenzene
 Normalized Internal Reference Method
I2(solution) = GI (Nsolvsolv + N ) 2 2 2
I2(solution)
GI Nsolvsolv 2
I2(solution)
I2 (solvent ) =  2
= 1 + N
Nsolvsolv 2
Slope =  2
Nsolvsolv 
 =
(Slope ×
Nsolvsolv)
1/2 2

25. Structure-NLO Property Relationship

26. Powder SHG by SHEW Method

27. X-Ray Structure of Tricyano-octupole

28. SHG = 40 x POM = 70 x LiNbO4 SHG = 7 x Quartz
1,3,5-Tricyano-2,4,6-tris(styryl)benzene
Crystal
Film
SHG = 40 x POM
= 70 x LiNbO4
SHG = 7 x Quartz
최근 화학의 관심 연구 주
Adv. Mater. 2005, 17, 196
Adv. Mater , 2107

29. 3-D Structure of Tricyano-octupole
최근 화학의 관심 연구 주

30. Spectral Measurements of Octupolar Crystal
최근 화학의 관심 연구 주
 Incident wavelength 1026 nm, IR intensity ~ 17 GW/cm2

31. Polarization Measurement I ( l = 987 nm )
최근 화학의 관심 연구 주
Crystal on its side :
q = p/2:
Polarization rotation
Sample plane X Z Y
=90°
f=88° =10°
---- IX
---- IY
SHG
2-Photon Fluorescence

32. What is Two Photon Absorption (TPA) ?

33. Application of TPA Optical power limiting
Three-dimensional imaging using two-photon laser scanning
Three-dimensional optical data storage
Photodynamic therapy
Two-photon excited fluorescence
image of Chromosome
3-D microstructure produced by
Two-photon-initiated polymerization

34. TPA Chromophores

35. Relationship between b and dTPA
(VB-3CT model)
J. Am. Chem. Soc. 2001, 123, 10658

36. Octupolar Molecules for TPA

37. Determination of TPA cross-section by PL method
Expression of the TPF signal Fi
Fi = K(i/2)niilI2
S / R = (KR /KS)(R/ S )(nR /nS)(FR /FS)
Ratio of TPA cross section for sample & standard
S/ R = (R/ S )(n R /n S )(F S /F R)(I R /I S)2
Things to consider
1. Difference in self-absorption
2. Identical optical setup
3. Refractive indices of solvents

38. Experimental Set Up for TPA(PL)
cell
Boxcar
Nd:YAG laser
OPO
355
Idler
LPF
Mirror
lens PD
PMT HV M
LPF : longwave pass filter, M : monochromator, PD : photo diode,
HV : high voltage power supply

39. One Photon and Two Photon Induced Fluorescence

40. Wavelength Dependence of TP Cross-Section for CN-Octupole

41. TPA & NLO Properties of Octupolar Molecules

42. Comparison with other TPA Chromophores
Table 1. NLO and TPA properties of 1,3,5-tricyano-2,4,6-tris(vinyl)benzene derivatives.
Comparison with other TPA Chromophores
a nm. b10-30 esu. c Corrected at  using a three-level model.2b d TPA cross-section measured at 800 nm by Z-scan method. e10-50 cm4s/photon. fTPA cross-section measured at 800 nm measured by fluorescence method. g The maximum TPA and the corresponding wavelength in the parenthesis.

43. Deeper penetration
Better 3D contrast
Less UV toxicity
Less Photo-bleaching
Live-cell imaging

44. Two-Photon Laser Scanning Microscopy (2PCLSM)

45. Top
Bottom 1 4 3 5 2
Z-scanning range : ~28m (5 sections, m step) OP TP

46. Lipid Rafts: Elusive or Illusive?
The lipid raft functions:
signal transduction
cholesterol homostasis
membrane traffficking
cytoskeletal organization
pathogen entry
neurodegenerative diseases
Too small to visualize it
A definitive proof of raft existence has yet to be obtained.
Sean Munro (2003) Cell, 115, 377

47. TPEF Images of A431 Cell ChemBioChem, 2007, 8, 553 Hydrophobic
(400~460nm)
Hydrophilic
(470~530nm)
ChemBioChem, 2007, 8, 553

48. Instrumentation  UV-VIS absorption, FT-IR, Raman, Fluorescence
 Femto-second, Nano-second time-resolved
laser spectroscopy (Pump-Probe, Up-conversion, TCSPC)
NLO measurement (Hyper-Rayleigh Scattering, TPF, Z-scan)
Confocal & Multi-photon Microscopy
Pressure generation : DAC(Diamond Anvil Cell)
Hydraulic Pressure Device
Computer Simulation
Quantum Mechanical- ab initio, semi-empirical.
Molecular Dynamics simulation

49. Dynamics! LS: light scattering HD: hydrodynamics
SAXS, SANS: small angle scattering
EM: electron microscopy
SMD: single molecular detection
FB: flow birefringence
FD: fluorescence depolarisation
EB: electric birefringence
LS: laser spectroscopy
Dynamics!

50. 신규 전략적 융합과학과제 제안 강신원 원장님(2005.11) 다차원 나노 비선형분광 및 생체 동역학
타 출연(연)과 차별화 및 타 지역 센터와 차별화 연구지원사업 육성
- 현재 종합적인 레이저 분광연구팀이 타 출연(연)과 기초(연)내에 없음
서울센터에서 연구 역량의 획기적인 강화 방안
- 대학의 연구능력을 활용하여 자체 연구능력의 향상과 연학 공동연구 기반 마련
연.학 협동 연구 과제의 활성화
전문사업단으로 발전을 지향하는 미래 지향적인 융합과학 과제 육성

51. 다차원 나노 비선형분광학 및 생체 동역학 (Multi-dimensional Nonlinear Spectro-Microscopy and BioDynamics )
비선형분광학( 물리.화학) + Microscopy(NT)
+ 생체물질.현상(BT)
- Challenge : 다차원 분광법(Multi-dimensional spectroscopy) 개발 (Solve challenging problems)
- Co-work/Research : Nonlinear Spectro-Microscopy for Bio-Materials & -Dynamics (high quality 논문)
- Instrumentation : Develop Nonlinear Spectrophotometer (주요 장비 개발)
전승준, 조민행, 김지환 참여 연구원

52. Protein Structure Determination: Tools and Comparison
Advantages and Limitations
 Researchers use a variety of tools to probe
protein function and interactions, with drug
discovery the major goal
 One of seven research fields
“Large-scale protein folding and 3-D structure studies”
Advantages
Restrictions
Molecular
crystal &
Low time-
resolution
X-ray
crystallo-
graphy
High spatial
resolution
Low time-
resolution
2D-NMR
Solution
M. Cho, Chem. Rev. 108, 1331 (2008)

53. exposures and fast enough framing rates for an event to be evaluated
High speed photography
“Recording optical or electro-optical information with adequately short
exposures and fast enough framing rates for an event to be evaluated
with a temporal and dimensional resolution which satisfies the experimenter”
Eadweard Muybridge (1887)

54. Molecular Motion Pictures
M. Cho, Nature 444, 431 (2006)

55. VIS optical diffraction limit
Structure
space (m)
Dynamics
time (sec)
10-9
10-6
10-3
10-12
10-15
XRD EM OM
NMR
MDS
TRS
VIS optical diffraction limit

56. 2D photon echo spectrometer

57. Nonlinear optical spectroscopy: Experimental setup
spectrometer
coh.
time
pop.
time
echo
time 1 2 3 4 t T t
spherical
mirror
4=LO 1 2 3
sig
sample
OD3 4
1&2 3
3&4 2 1
diffractive
optic (DO)
delay 2
delay 1

58. 2D photon echo experimental scheme

59. Multidimensional Spectroscopy Laboratory (MDSL)

60. Multidimensional Spectroscopy Laboratory (MDSL)
H-bond
making
H-bond
breaking kb 60

61. Femtosecond CD and ORD spectrometer (novel technique)

62. Conventional CD

63. Theory Brief Report Full Report
Theory : THE JOURNAL OF CHEMICAL PHYSICS 2008, 129,
Femtosecond spectral interferometry of optical activity: Theory
Hanju Rhee, Jeong-Hyon Ha, Seung-Joon Jeon, and Minhaeng Cho
Letters : Nature 2009, 458, 310
Femtosecond characterization of vibrational optical activity of chiral molecules
Hanju Rhee, Young-Gun June, Jang-Soo Lee, Kyung-Koo Lee, Jeong-Hyon Ha, Zee Hwan Kim, Seung-Joon Jeon & Minhaeng Cho
(Nature News & Views : “Handedness in quick time” 2009, 458, 289)
Full Paper : J. Opt. Soc. Am. B 2009, 26, 1008
Phase sensitive detection of vibrational optical activity free-induction-decay: vibrational CD and ORD
Hanju Rhee, Young-Gun June, Zee Hwan Kim, Seung-Joon Jeon, and Minhaeng Cho

64. Theory :
CD :
ORD :
Femto-second Laser pulse
Coherently emmitted free-induction-decay(FID)
Cross-polarization detection scheme
(Calcite plate polariser, , μm,extinction ratio )
Heterodyne-detected Fourier-transform
spectral interferometry(FTSI)

65. 주요 해결 방안

66. P

67. H. Rhee, Y. -G. June, J. S. Lee, K. -K. Lee, J. -H. Ha, Z. -H. Kim, S
H. Rhee, Y.-G. June, J.S. Lee, K.-K. Lee, J.-H. Ha, Z.-H. Kim, S.-J. Jeon, M. Cho
NATURE, 2009,458, 310

68. Detailed experimental schemes and
Results
Femtosecond
spectra
interferograms
Vibrational
circular
dicrhoism
Vibrational
optical
rotatory
dispersion 68

69. Time-resolved CD and ORD in femtosecond timescale!69

70. Femtosecond Difference Frequency Spectroscopy
Novel Vibrational Optical Activity Measurement Technique

71. Ultimate goals

72. 생명 현상의 이해 분자수준에서의 구조-활성 연관성 연구
독립된 생체분자의 구조/동역학 연구
수십나노미터 수준의 생체 군집의 구조/활성
비선형 다차원 분광학의 이론 및 실험적 연구
화학선택적 이미징이 가능한 분광학적 현미경 실험
Multidimensional spectroscopy with time-space resolution
Biostructure and Biodynamics 72

73. Chemical exchange and reaction dynamics protein aggregation process
In summary
       다차원 분광법에 대한 실험법과 데이터 해석법의 확립을 통하여 단백질,
핵산과 같은 생화학물질의 구조와 동역학 연구법 개발 
Chemical exchange and reaction dynamics
protein aggregation process
Protein folding
DNA and RNA dynamics 73

74. Molecular Motion Pictures AGAIN!
Novel Spectroscopic
Techniques
(KBSI-Seoul Center)
2D IR spectroscopy
fs CD and ORD
Experimentally demonstrated
3. fs optical activyt-DFG
New proposal
M. Cho, “Spectroscopy, Molecular Motion Pictures”, Nature 444, 431 (2006)
M. Cho, “Coherent two-dimensional optical spectroscopy, Chem. Rev. 108, 1331 (2008)

75. Long term projects

76. What is next? Dynamics in cell : spatial & time resolution
(Explain life by physical & chemical principles?)
Pair potential, Stochastic ?
Folding, Docking
Origin of life
New force – Casimir force, Brownian Ratchet
예 :TR Nanoscopy : overcome diffraction limit
STED(S. Hell), PALM/STORM(?)

77. Co-worker Spectroscopy (전승준) Theory & Exp (조민행) Synthesis (조봉래)
Raman, HRS (김동호, 연세대)
Time-resolved spec (정세채, 표준연)
(주태하, 포항공대)
Crystal, Film NLO (J. Zyss, France)
Financial Support
NRL-MOST
CRM-KOSEF
KBSI