X-ray imaging of bio-matter @ synchrotrons & lab sources Janos - - PowerPoint PPT Presentation

X-ray imaging of bio-matter @ synchrotrons & lab sources

Janos Kirz

EuXFEL 11/2017

Origins

• W. C. Röntgen 1895

– Nov 8 – first observation – Nov 9 – Dec 27 experimentation, write-up – Dec 28 – manuscript submitted

• Sitzungsberichte der Physikalischen-medizinischen Gesellschaft zu Würzburg
• See bones in hand – shadowgraphs

Instant sensation around the world Radiology – absorption contrast Works well for bone fracture air vs fluid in lung tooth decay

3D Imaging: CAT scans

• 3D imaging based on many projections
• 1979 Nobel prize Godfrey Hounsfield

& Allan McLeod Cormack

• Resolution: several mm
• Limitations: breathing, beating heart, digestion,…

Better radiology ?

• Hundreds of images: high radiation dose!
• Absorption contrast in soft tissue poor
• To reduce dose – improve contrast!
• Extract phase shift from interference pattern

– Bonse-Hart interferometer

• Appl. Phys. Lett 6, 155 (1965)

– Phase contrast tomography

• A. Momose et al.

– Nature Med. 2, 473 (1996)

Interferometry without crystals

• Grating-based phase measurement –

– Using Talbot (1836) effect (self-image)

• A. Momose et al. Jpn. J Appl. Phy. 42, L866 (2003)
• Spring-8 source for coherence
• Object distorts Moire pattern
• Talbot-Lau interferometer
• Third grating for use with ordinary X-ray tube

– F. Pfeiffer et al. Nature Phys. 2, 258 (2006)

• Intensive development worldwide

Absorption & Phase contrast

• F. Pfeiffer et al. Nature Phys. 2, 258 (2006)

Application to mammography

(From Konica – Minolta) Phase contrast Absorption contrast

X-ray micro-Tomography-ALS 8.3.2

Lots of Math

Microtomography

• Simple projection onto detector

Resolution: ~ 1-5 µm

• Commercial “microscopes”
• Lab sources or synchrotrons

– Zeiss (formerly XRADIA) – Brucker (formerly Skyscan)

• 3D Studies of bone structure,

seeds, small animals

Examples of Grapevine Xylem

11

Get insights in insect flight control

Investigate the biomechanics underlying flight manoeuvres and gaze shifts à CT following the dynamics of 100+ Hz wing beat!

Need:

• single-shot propagation-based phase contrast
• high-speed X-ray tomographic microscopy

Requires: Coherence and Flux BRIGHTNESS So, this is a perfect task for a synchrotron!

SLS beam

• S. M. Walker et al. P{LOS Biology 12, e1001823 (2014)

Toward higher resolution

• Microscopes with zone plate optics

– Resolution ~ 20 nm (50 nm in 3D)

• Radiation damage becomes limitation!
• Cryo preserves morphology
• Instruments at

– BESSY II – ALS – ALBA – …

Cryo Soft X-ray Tomography of Cells at the MISTRAL beamline

reconstructed slice

1 µm

• E. Pereiro et al. J. Synchr. Rad. 16, 505-512 (2009)
• A. SorrenBno et al. J. Synchr. Rad. 22, 1112-1117 (2015)

Radiation damage in biological samples Frozen hydrated state of protein Howells et al. JESRP 170, 4 (2009) Inverse fourth power law of dose vs resolution: Dose ~ 1/resolution-size4

Resolution limit ?

The ultimate challenge

15

Scanning X-ray diffracBon microscopy

• P. Thibault, M. Dierolf, A. Menzel, O. Bunk, C. David, F. Pfeiffer, Science, 321, 379-382 (2008).

Ptychography with a focused X-ray probe Pilatus 2M

• J. Rodenburg

Reconstruct bith amplitude and phase

Locate, extract and further study intact hallmarks of Parkinson disease Large volumes High resoluBon As close to naBve state as possible (no staining)

Quickly biopsy-punched, infiltrated with cryo protectant, mounted on pin and gradually frozen Trimmed with cryo ultramicrotome Cryo transferred to OMNY ~ 80 micron at the base, ~ 100 nm 3D resoluBon 446 projecBons 17 hour measurement

Frozen hydrated unstained brain tissue

Page 16

Sarah Shamoradian Shamoradian et al., Sci. Rep. 6291 (2017)

3D Elemental mapping by XRF

• 3D scan as for tomography
• Record fluorescence spectrum for

each point

• Perform tomographic recontstruction

for each element

3D elemental microtomography of Cyclotella meneghiana

• M. de Jonge, et al., PNAS 107, 15676, (2010)

Changing Landscape of X-ray facilities

• ~ 1985- dedicated storage rings

– 30+ years: a revolution in X-ray analysis

• ~ 2010 Toward higher brightness, coherence

– MBA lattice, MAX IV, Sirius, storage ring upgrades – FELs LCLS, FLASH, FERMI, SACLA, EuXFEL,…

• Older sources: DORIS, NSLS, Daresbury,… shut

down

– Fewer stations available for routine measurements

New initiatives in Lab-scale sources – to fill the gap

• EXCILLUM – liquid metal jet
• Lyncean – back-scattered Compton
• Sigray – microstructured anode

in diamond substrate

Conclusions I

• X-rays are great!

– Penetrate opaque objects – Rich spectra allow elemental & chemical info

• Radiation damage is a concern
• One way to mitigate radiation damage: cryo
• Other ways:
• many copies, as in crystals
• or diffract & destroy
• ... but ptychography not compatible

Conclusions II

• A bit of humility:
• There is competition!

– MRI for imaging humans – Cryo EM for high resolution on thin samples – Super resolution visible light microscopy

Acknowledgments

• Thanks to colleages who provided

material for this talk:

– Manuel Guizar Sicarios – Eva Pereiro – Marco Stampanoni – Andrew McElrone

Thank you