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CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi [email protected] tional Symposium on Molecular Spectroscopy 62nd Meeting - June 18-

CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

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Basic Principles of Optical Tweezers Light besides energy carries Lineer momentum Force Classical : EM theory, Radiation pressure Quantum : Photon momentum, Angular momentum Torque

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Page 1: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

CONSTRUCTION OF A RAMAN TWEEZER SYSTEM

FOR INVESTIGATION OF BIOLOGICAL MOLECULES

Günay Başar

İstanbul Teknik Ü[email protected]

International Symposium on Molecular Spectroscopy 62nd Meeting - June 18-22, 2007

Page 2: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

• Basic principles of optical tweezers• Experimental set-up• Advantages of Raman Tweezer set-up• Test measurements on micro particles

and living cells

Outline

Page 3: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

Basic Principles of Optical Tweezers

Light besides energy carries

Lineer momentum Force Classical : EM theory, Radiation pressure

Quantum : Photon momentum,

Angular momentum Torque

dP Fdt

hpc

dLdt

Page 4: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

Radiation Presure→ Optical Trapping

1 mW

1μm F ~10 pNm ~pg

F ~ 10 pN

a ~105 g

Page 5: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

a b

Fb Fa

F

b a

a b

b a

a b

FFa Fb

f

f

fFa

Fb

F

b

a

La se r Be a m

M ic . O b je c tive

PinP o ut

Page 6: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

(A. Ashkin, Biophys. J. (61), 1992)

21

2

21

2

cos(2 2 ) cos 21 cos 2

1 2 cos 2

sin(2 2 ) sin 2sin 2

1 2 cos 2

z scat

y grad

T r Rn PF F Rc R R r

T r Rn PF F Rc R R r

Single Ray SolutionP : PowerMomentum per unit time:

1n Pc

P

r

O

y

zPT2

PTR2

PTR2 2

PRn1

n2

Page 7: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

Be a m

f

r

m a x

S

rm a x

x

y

z z

y

w

O

Be a mr rm a x

w

z

f

Be a m a xis

Fg S

Fs

O

M ic .o b je c tiveb a c k a p e rture

Extended Laser Beam

Page 8: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

• Large NA objectives• Overfilling• n2 > n1 • Power ~1mW is enough to trap

Critical Points for Optical Trapping

Page 9: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

Experimental setup

Page 10: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International
Page 11: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

1µm trapped polystyrene particle

Page 12: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

4.6 µm trapped polystyrene particle

Page 13: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

~ 5 µm trapped yeast cell

Page 14: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

~ 5 µm trapped yeast cell

Page 15: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

• The ability to get Raman Spectrum of a single living object in their natural environment.

• Decreasing of background signals from cover slip.

• No mechanical immobilization of living cells.

Advantages of Raman Tweezer Set-up

Page 16: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

Excitation wavelength: 780 nmPower: 6 mW on the sampleAccumulation: 100 s

Raman spectrum of trapped polystyrene particle

Page 17: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

Raman spectrum of trapped yeast cell

C. Xie et al. Opt. Exp. 25, 6208-6214 2004

Page 18: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

Collaboration

Istanbul University

Physics Department

Prof.Dr. Sevim Akyuz

• Optimizing Laser Tweezer Raman Spectrometer• Investigation of interaction between cell membranes and their environment

Future Work

Support by the Scientific and Technological Research Council of Turkey under the project number 105T482 is gratefully acknowledged.

Our Laser Spectroscopy Groupat Istanbul Technical UniversityPhysics Engineering Department

PhD studentSeda Kin

MsC studentUğur Parlatan

BsC studentsOnur PusulukBerkin Malkoç

Group leaderAssoc.Prof. Dr. Gunay Basar

Page 19: CONSTRUCTION OF A RAMAN TWEEZER SYSTEM FOR INVESTIGATION OF BIOLOGICAL MOLECULES Günay Başar İstanbul Teknik Üniversitesi International

Thank You!