National resource for high-National resource for high-field NMR field NMR imaging imaging and and spectroscopyspectroscopy

Focus on advanced Focus on advanced basicbasic and and clinicalclinical applications applications and technology and technology development development

““Biological-biomedical arm” Biological-biomedical arm” of of National High Magnetic National High Magnetic Field Lab (NHMFL)Field Lab (NHMFL)

MBI-UF Advanced Magnetic ResonanceMBI-UF Advanced Magnetic ResonanceImaging and SpectroscopyImaging and Spectroscopy

(AMRIS) Facility(AMRIS) Facility

http://www.mbi.ufl.edu/facilities/amris

136 Mhz, 3.0 Tesla, 60 cm horizontal bore136 Mhz, 3.0 Tesla, 60 cm horizontal boreMRI/S of live animals (humans, primates, dogs, MRI/S of live animals (humans, primates, dogs, etcetc.).)

136 Mhz, 3.0 tesla, 80 (136 Mhz, 3.0 tesla, 80 ( 94) cm horizontal bore 94) cm horizontal boreMRI/S of live humansMRI/S of live humans

200 Mhz, 4.7 tesla, 33 cm horizontal bore200 Mhz, 4.7 tesla, 33 cm horizontal boreMRI/S of live animals (cats, rabbits, rats, mice, MRI/S of live animals (cats, rabbits, rats, mice, etcetc.).)

473 Mhz, 11.1 tesla, 40 cm horizontal bore473 Mhz, 11.1 tesla, 40 cm horizontal boreMRI/S of live animals (primates, cats, rabbits, rats, mice, MRI/S of live animals (primates, cats, rabbits, rats, mice, etcetc.).)Solution state NMR spectroscopy of biomolecules Solution state NMR spectroscopy of biomolecules (multiple (multiple samplessamples))

500 Mhz, 11.7 tesla, 5.2 cm vertical bore500 Mhz, 11.7 tesla, 5.2 cm vertical boreSolution/solid state NMR spectroscopy of biomoleculesSolution/solid state NMR spectroscopy of biomolecules

600 Mhz, 14.1 tesla, 5.2 cm vertical bore600 Mhz, 14.1 tesla, 5.2 cm vertical boreSolution state NMR spectroscopy of biomoleculesSolution state NMR spectroscopy of biomolecules

Cryoprobe to boost S/N by a factor of 4Cryoprobe to boost S/N by a factor of 4MRI/S of superfused cells/tissuesMRI/S of superfused cells/tissues

750 Mhz, 17.6 tesla, 8.9 cm vertical bore750 Mhz, 17.6 tesla, 8.9 cm vertical boreMRI/S of superfused cells/tissues & of live animals (MRI/S of superfused cells/tissues & of live animals (e.ge.g., mice)., mice)Solution/solid state NMR spectroscopy of biomolecules Solution/solid state NMR spectroscopy of biomolecules (multiple (multiple samples)samples)

Cryoprobe under developmentCryoprobe under development

MBI-UF AMRIS InstrumentationMBI-UF AMRIS Instrumentation

                                             

MBI-UF AMRIS: From Molecules to ManMBI-UF AMRIS: From Molecules to Man

2.53.03.54.04.55.05.5 ppm

5.165.185.205.225.245.265.285.305.32 ppm

Animal MRI/MRS

High-Resolution Structural Biology Microsample (1.5l) spectroscopy

MR Microscopy (ex vivo)

Single cell MRI/NMR

Human research

MBI-UF AMRIS RF Engineering LabMBI-UF AMRIS RF Engineering Lab

coil

Ct

Microcoils and arrays (MRI & MRS/NMR) Superconducting probes

Phased array coils Large volume/High frequency Human coilsBeck Beck et al.et al. (2002) (2002) MAGMAMAGMA 13: 152-157 13: 152-157

MBI-UF AMRIS: 2002 User Research MBI-UF AMRIS: 2002 User Research HighlightsHighlights

Brian Shilton (Univ of Western Ontario), Hargrave, Smith, McDowell, and Edison, “High-field structural studies of Rhodopsin/Arrestin complexes”

Elisar Barbar (Ohio University) and Edison, “Structural biology of microtubule transport”

Cottrell (St. Andrews), Zachariah, Dossey, Edison, “3D structure of a neuropeptide bound to its receptor”

Webb (Illinois), Thelwall, Grant, Blackband, “NMR Microscopy of a Single Neuron Isolated from Aplysia Californica”

Grant, Plant, Mareci, Blackband, Webb (Univ. Illinois), Aken (Univ. Arizona), Grant, Plant, Mareci, Blackband, Webb (Univ. Illinois), Aken (Univ. Arizona), ““Proton Spectra from a Single Neuron Isolated from Proton Spectra from a Single Neuron Isolated from Aplysia Californica”Aplysia Californica”

Benveniste (Brookhaven Nat. Lab), Zhang (Brookhaven), Grant, Blackband, “MR Microimaging Studies of Mouse Brains For Generation of a Web Based Atlas and Methods for Identification of Brain Structures”

Silver, Plant, Blackband, Benveniste (Brookhaven Nat. Lab), “Normal Mouse Brain Normal Mouse Brain MRI MRI In Situ”In Situ”

Webb (Illinois), Zhang (Illinois), Edison, “Double Protein NMR coil”

Funding for AMRIS provided by:Funding for AMRIS provided by:

Thank Dr. Stephen Thank Dr. Stephen Blackband for providing Blackband for providing the slices abovethe slices above

High Field MR Technology Development

Yu LI

McKnight Brain InstituteAdvance Magnetic Resonance

Imaging and Spectroscopy FacilitiesUniversity of Florida, Gainesville, FL 32610

Outline

Research Background Basic MR Principles Small-Volume Protein NMR MR Parameters Estimation Imaging Technology Summary

Roadmap

Research Background Basic MR Principles Small-Volume Protein NMR MR Parameters Estimation Imaging Technology Summary

History

1946 MR phenomenon – Bloch & Purcell

1952 Nobel Prize/Physics – Block & Purcell

1955 NOE Effect – Solomon

1966 Fourier transform NMR – Ernst, Anderson

1973 Backprojection MRI – Lauterbur

1975 2D NMR – Jeener, Ernst; Fourier Imaging – Ernst

1980 MRI demonstrated – Edelstein

1985 Solution structure of small protein – Wüthrich

1986 Gradient echo imaging; NMR microscope

1987/8 3D NMR + 13C, 15N isotope labeling

1989 Echo-planar imaging

1991 Nobel Prize/Chemistry – Ernst

1996/7 NMR development in maromolecular structure determination; Anisotropic diffustion

2002 Nobel prize/Chemistry – Wüthrich

MR Research Areas

MR Spectroscopy– Solution state– Solid state

MR Imaging– Human/Animal imaging– Microimaging– Material imaging

Data Processing– Spectral data processing– Image reconstruction– Image post-processing

High Field MR Technology

NIH Resource Resource Cores:

– High Field Small Animal Imaging

– Microimaging and Microspectroscopy

– High-sensitivity and High-throughput Solution State NMR

Roadmap

Research Background Basic MR Principles Small-Volume Protein NMR MR Parameters Estimation Imaging Technology Summary

MR Phenomena: Resonance

Michael Sattler EMBL Heidelberg, Biomolecular NMR Structure, http://www.EMBL-Heidelberg.de/nmr/

B0

0Larmor frequency: , geromagentic ratioB

1 pB

1 p1

cos( ) 0<t<( )

0 else

B tB t

0Equilibrium state: M M z

B0

MR Phenomena: Free Relaxation

1 0 01

relaxation: ( ) (0) exp( )z z

tT M t M M M

T

22

relaxation: ( ) 0 expxy xy

tT M t M

T

Mz

Mxy

M

x

y

z

x

y

MR Signal: FID

/

1

( ) cosi

nt T

i i ii

s t Ae t

MR Parameters Factors MR Signal

i Microrscopic environment Frequency shift

Ai Nucleus spin density and Object volume

IntensityTi Physiological or physicochemical properties

i Molecule mobility

Nuclei of MR Interest

Nuclei  Net Spin  (MHz/T) 

1H 1/2  42.58 

2H 1  6.54 

31P 1/2  17.25 

23Na 3/2  11.27 

14N 1  3.08 

13C 1/2  10.71 

19F  1/2  40.08 

MR Application

Michael Sattler EMBL Heidelberg, Biomolecular NMR Structure, http://www.EMBL-Heidelberg.de/nmr/

Fourier Transform

Image Reconstruction

MR Instrumentation

Magnet

RF coil

and

ObjectG

radi

ent c

oil

Gradient coil

Transmitter Receiver

Synthesizer ADC

Console

Advantage: Information Rich

Molecule structure Anatomical structure Physiological mechanism Pathophysiologies Biological functional structure

Drawback: Low SNR

Spectroscopy– Low sample efficiency

– Low throughput

Imaging– Long imaging time

– Low resolution

High Field TechnologyHigh Field Technology

Roadmap

Research Background Basic MR Principles Small-Volume Protein NMR MR Parameters Estimation Imaging Technology Summary

Protein Structure

Primary Secondary Tertiary Quaternary

Amino Acid Chain structure

Protein NMR

Michael Sattler EMBL Heidelberg, Biomolecular NMR Structure, http://www.EMBL-Heidelberg.de/nmr/

Structure Information

Frequency shift: chemical structure dependence Spectral peak structure: connection between

different chemical groups

Frequency shiftFrequency shift

/

1

( ) cosi

nt T

i i ii

s t Ae t

Small Volume / High Field

Significance of small volume– Time of sample preparation– Expense– Availability

High field rationale

120 s

noise

BiSNR V

V

D.I.Hoult and R.E.Richards, J.Magn.Reson, 24, 71-85 (1976)

BB00 field field

RF coil design RF coil design 1

noise

Bi

V

0

Current Probe Technology

Required sample volume: 600 µL

Saddle and Solenoid

B1

Current: i Current: i

D.I.Hoult and R.E.Richards, J.Magn.Reson, 24, 71-85 (1976)

“the disappointing signal-to-noise ratio experienced with superconducting system is a direct consequence of the use of saddle-shaped coils”

SaddleSaddle SolenoidSolenoid

Solenoid Probe Design

1H

Lock

15N

Solenoid Coil

C1

L1 C2

C3

C4

C6

C9 C10

L4 C8

C13C12

L2

C5

L3 C7

13C

L5C11

C15C14

Experimental Comparison

Solenoid Probe Commercial Saddle Probe

Sample Volume

60 µL 600 µL

SNR 97 91

Roadmap

Research Background Basic MR Principles Small-Volume Protein NMR MR Parameters Estimation Imaging Technology Summary

MR Parameters in Frequency Domain

212

( )1 ( )

ni

i ii

AS

T

/

1

( ) cosi

nt T

i i ii

s t Ae t

Fourier Transform

LinewidthIntensity

Frequency

CE NMR

B(f)

Noise

Se(f) Seb(f)

SNR = 36.6

SNR = 22.0

Problem Formulation

B(f)

Noise

Sr(f) Srb(f)

B(f)

Noise

Se(f) Seb(f)

Know Sr(f), Detect Srb(f), Estimate B(f)

Know B(f), Detect Seb(f), Estimate Se(f)

Gradient Decent Method

Srb(f)

Sr(f) +

_ eb(f)B(f)

(•)2

b

[ 1] [ ] bPara n Para nPara

Seb(f)

B(f)+

_ es(f)Se(f)

(•)2

s

[ 1] [ ] sPara n Para nPara

Gradient Decent Method

Error function

ParametersOptimum Values

Multiresolution detection with wavelet

Low resolution / Low SNR

High resolution / High SNR

Wavelet transform

Scale decrease

S. Mallat, and W.L. Hwang, IEEE Trans. on Information Theory, Vol. 38(2), 617-643 (1992).

Resolving Results

100 mM sucrose in D2O

Roadmap

Research Background Basic MR Principles Small-Volume Protein NMR MR Parameters Estimation Imaging Technology Summary

MR Signal Intensity

MR Parameters Factors

Ai Proton density

Ti Physiological or physicochemical environment

i Molecule mobility

Image Contrast: MR Parameters-weighted

Proton density– Physical composition

T1

– Soft tissue

T2

– Tissue structure– Tissue metabolism– Pathophysiologies

Image Contrast: MR Parameters-weighted

T2*

– Vascular physiology– Biological functions

Apparent Diffusion Coefficient (ADC)

– Tissue microstructure– Tissue composition– Tissue constitutes– Architectural organization

0bDS S e

3D Brain / Spinal Cord Imaging

T2-weighted Images of rat brain and spinal cordHigh resolution: below 40 µm (17.6T)

B. Beck, D.H. Plant, S.C. Grant, PlE. Thelwall, X. Silver, T.H. Mareci, H. Benveniste, M. Smith, S. Crozier, S.J. Blackband

Brain Slice Imaging

Diffusion weighted microimage of rat brain sliceHigh Resolution: 20 µm (14.1 T)

S.J. Balckband, J.D. Bui, D.L. Buckley, T. Zelles, H.D. Plant, B.A. Inglis, M.I. Phillips

Neuron Cell Imaging

Diffusion-weighted images of a single neuron cellCytoplasm (C) and nuclear (N) in artificial sea water (S).High Resolution: 20 µm (14.1 T)

S.C. Grant, D.L. Buckley, S. Gibbs, A.G. Webb, and S.J. Balckband

Roadmap

Research Background Basic MR Principles Small-Volume Protein NMR Spectral Resolution Restoration Imaging Technology Summary

High Field MR Technology

Hardware development– Magnet– Coil geometry / dimension– RF circuit design

Algorithm development– MR parameters estimation– Biomedical information and MR parameters– Image processing– EM field calculation

Acknowledgement

Drs Arthur Edison

Andrew Webb

Stephen Blackband

Samuel Grant

Jim Roca

Paul Moliter

William Brey

Feng Lin

Peter Gor’kov

Jim Norcross

Terry Green