Slide: 1
Contact-free manipulation and probing of single biological and soft matter objects
Christian Riekel
European Synchrotron Radiation Facility, Grenoble, France
Overview on selected contact free techniques
Scope of applications
MicroSAXS/WAXS cameras
Contact free techniques: applications & challenges
XDL 2011 - 27&28 June- Christian Riekel
Slide: 2
ESRF-ID13 beamline team Manfred Burghammer et al.,Silvia Santucci (optical tweezers)Rita Graceffa (inkjet systems; now at APS BioCAT)Angelo Accardo (superhydrophobic surfaces)
Italian Institute of Technology (Genoa-Italy)Enzo Di Fabrizio et al., (nanotechnology)
Laboratorio TASC (Trieste-Italy)Dan Cojoc (optical tweezers)Institute of Biophysics and Nanosystems Research (Graz-Austria)Heinz Amenitsch (optical tweezers)
Acknowledgements
XDL 2011 - 27&28 June- Christian Riekel
EEC funding
Slide: 3
Contact-free and quasi contact-free manipulationOptical Tweezers Inkjet Superhydrophobic Surface
piez
o
gradient force scattering force
focused laser beam
stroboscopic display
XDL 2011 - 27&28 June- Christian Riekel
Particles in capillary
Slide: 5XDL 2011 - 27&28 June- Christian Riekel
Nano-grippers (NanoHand project)
10 m
HeLa cells @ r.t.
50 m
• Avoid deformation of ″soft″ objects by ″contact″ forces• Avoid surface induced processes and shearing effects by walls
Scope of applications
• Reduce sample volumes!
• Single objects: positioning, manipulation & assembly• Probing of objects in functional states, often in aqueous environments• Raster probing: heterogeneity & distribution of radiation damage
Slide: 6
focusing optics
dete
ctor
XDL 2011 - 27&28 June- Christian Riekel
reciprocal space order resolution ()
real space raster-increment (focal spot)
Resolution
source
sample
F2F1 F2
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focusing optics
dete
ctor
XDL 2011 - 27&28 June- Christian Riekel
S’h/v Rad; fwhm)
sample
Br13 keV ~2*1021 (upgrade)
undulator source parameters
source
S,S’h,v
F2F1 F2
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focusing optics
dete
ctor
+
XDL 2011 - 27&28 June- Christian Riekel
Sh/v (m; fwhm)S’h/v Rad; fwhm)
high-: ID02 (high resolution SAXS)ESRF 6 GeV
sample
Br13 keV ~2*1021 (upgrade)
undulator source parameters
source
S,S’h,v
F2F1 F2
Slide: 9
focusing optics
dete
ctor
XDL 2011 - 27&28 June- Christian Riekel
S’h/v Rad; fwhm)
undulator source parameters
high-flux nanofocusCHESS-ERL 5 GeV
high-coherence21 m fwhm
5 Rad fwhm
sample
~9*1022Br13 keVBilderback et al., NJP (2010) 12, 035011
~2*1021 (upgrade)
source
S,S’h,v
F2F1 F2
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10-110-210-3
109
108
107
106
105
104
103
102
101
ESRF-ID02
10-4
XDL 2011 - 27&28 June- Christian Riekel
ERL-5m ERL-1m ERL-0.1m?
I(Q
)
148 nm SiO2 spheres
ESRF: ID02 (high-)
ID13 (low-)
ERL pinhole SAXS
13 keV, Si111
ID02 upgrade ID13~10 m 1-0.1 m
ERL SAXS camera for ultrasmall sample volumes
≤800*200 m2
• self assembly: actin…• transient processes: flames…• biomimetic systems: biomineralization…• mesoscopic processes: active matter…
COLLECTIVE PROPERTIES
INDIVIDUAL PROPERTIES• single objects: cells, fibrils…• local heterogeneity
Brilliance and SAXS pinhole camera
Q (nm-1)60 nm600 nm
CC
D
Slide: 12
source&optics detector
2d
2s )/()/( 2D
22d LPSFb /)(
=0.1 nm PSF= 30 m
LD (mm)
s
(nm
-1)
Order resolution - s
~1 mrad
~100 rad
ERL microSAXS/WAXS pinhole camera
100 nm focus
1 m focus
F2= 226 mmF1=48 000 mm
on-axis refractive optics
ERL-undulatorhigh-coherence
10 100 1000
1E-3
0.01
XDL 2011 - 27&28 June- Christian Riekel
LD
0.2 nm
1844
pix
els
b
sample<15
0 m
5.3 Rad21.2 m
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Ashkin, PRL (1970) 24, 156 Ashkin et al., Opt. Lett. (1986) 11, 288
light guide
ESRF optical tweezers setup
Optical tweezers: 10-10 – 10-13 N forces
mirror
illumination
CCD
IR laser
objective
Spatial Light Modulator
Santucci et al., Anal. Chem. (2011) 83, 4863
capillary&sample
XDL 2011 - 27&28 June- Christian Riekel
Slide: 14
Ashkin, PRL (1970) 24, 156 Ashkin et al., Opt. Lett. (1986) 11, 288
Optical tweezers: 10-10 – 10-13 N forces
mirror
illumination
CCD
IR laser
objective
Spatial Light Modulator
capillary&sample
XDL 2011 - 27&28 June- Christian Riekel
SiO2 microbeadsPadgett group Univ. St. Andrews UK
Slide: 15
Ashkin, PRL (1970) 24, 156 Ashkin et al., Opt. Lett. (1986) 11, 288
Optical tweezers: 10-10 – 10-13 N forces
mirror
illumination
CCD
IR laser
objective
Spatial Light Modulator
Santucci et al., Anal. Chem. (2011) 83, 4863
capillary&sample
XDL 2011 - 27&28 June- Christian Riekel
20 m
100 K
Room temperature PXradiation damage evolution?
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ba 100 m 2,2,0
3,1,0
1,3,0
2,0,0
0,4,0 1,3,0
1,1,0 +4,2,0
3,3,0
2,4,0b* a*1,1,0
1,1,0
(hk0) planea=b=7.5 nm
50 ms/point
averaged raster-scan
10
50raster-scan of 110 reflection across crystal
Inte
nsit
y(a
.u.)
50 ms 100 ms 150 ms 200 ms
0.25 nm resolution limitview along beam1 m beam–13 keV5 m raster-steps
XDL 2011 - 27&28 June- Christian Riekel
Santucci et al., Anal. Chem. (2011) 83, 4863
Insulin crystaltrapped in capillary
Optically trapped insulin crystal
Slide: 17XDL 2011 - 27&28 June- Christian Riekel
Optical tweezers challenges
• Protein microcrystals and serial crystallography
• Radiation damage studies
• Small soft and biological objects
• Aggregation, fusion, reaction…
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microscope image of cluster of ~10 liposomes in capillary
Cojoc et al., APL (2007) 91, 223107
~1 mm
m
raster-diffraction image5.3 nm
1 m beam
multiwalled liposome,phospholipid membranes
peptide induced liposome fusionNomura et al., PNAS (2004) 101, 3420
0 (sec)3.6 4.2 4.8 5.4 6.6 7.8 10.8
5 m
XDL 2011 - 27&28 June- Christian Riekel
MD simulation (Stevens, Sandia Lab)
Challenge: probing of single liposome
Optically trapped liposomes
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10 s 2*104 microdrops/point
stroscopic SAXS1 m beam-8 m raster
Inkjet systems generate wall-free reaction volumes
...5 µs pixel detector frames
triggering sequence500 s strobe period
8 m8 m
XDL 2011 - 27&28 June- Christian Riekel
ballistic (2m/s) water microdropsdrop-on-demand inkjet system268 picoliters – 80 m stroscopic display
Graceffa et al., APL (2009) 94, 62902Rita Graceffa, PhD thesis, Grenoble (2010)
MEDIPIX: 256x256 pixels
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10 s 2*104 microdrops/point
stroscopic BioSAXS1 m beam-center drop
Inkjet systems & BioSAXS
XDL 2011 - 27&28 June- Christian Riekel
ballistic (2m/s) water microdropsdrop-on-demand inkjet system268 picoliters – 80 m stroscopic display
Graceffa et al., APL (2009) 94, 62902Rita Graceffa, PhD thesis, Grenoble (2010)
Cytochrome C
...5 µs pixel detector frames
triggering sequence500 s strobe period
MEDIPIX: 256x256 pixels
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0 ms
2 ms
4 ms
bufferunfolded cytochrome C @ pH2
folded cytochrome C
fold
ing
tim
eOH-
20µm*20µm
buffer
10*10µm2
10*10µm2
Rita GraceffaPhD thesis, Grenoble (2010)
XDL 2011 - 27&28 June- Christian Riekel
Inkjet systems & BioSAXS
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protein microarray by inkjet technologyLaurell lab - Lund University
Inkjet challengesMicrodrops
• In flight Fewer and smaller microdrops in stroboscopic sequenceSingle microdrop scattering at ERL?BioSAXS with
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non-wetting plant leave artificial non-wetting surface(superhydrophobic)
Quasi contact-free superhydrophobic surfaces
XDL 2011 - 27&28 June- Christian Riekel
Accardo et al., Langmuir (2010) 26, 15057
PMMA
Slide: 24XDL 2011 - 27&28 June- Christian Riekel
Raster-diffraction
Hauser et al., PNAS (2011) 108, 1361
Accardo et al., Soft Matter, ASAP, 13.6.2011
100 m
Lysozyme fibrillation3-mer peptide residue
Blake et al., Structure (1996) 4, 989
SEM
Peptide and protein precipitation
-sh
eet
helix
amyloid
fibr
il
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aggregation Nbr of peptide molecules
Nbr
inte
rcha
inhy
drog
en b
onds
Peptide aggregation simulation
Auer et al., HFSP Journal (2007) 1, 137
-sheet aggregation
fibroin
I Q (n
m-1)
aggregate102
101
100
10-1
10-2
10-3fibroin
in solution
BioSAXS: silk fibroin aggregation
0.1 1
260
nmag
greg
atio
n
Q (nm-1)
Martel et al., JACS (2008), 130, 17070
Microfluidic cell ml-scale liquid consumption
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• BioSAXS pL - nL drops, m to subm beams
• Faster scans continuous scan with subms patterns, kinetics
• Coherent imaging complementary approach, in situ processes…
• Stability avoid sample movements during fast scans
Superhydrophobic surfaces challenges
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Pixe
l det
ecto
r
XDL 2011 - 27&28 June- Christian Riekel
beam steering optics
ERL-undulator
x/y/zrot
scanning-gonio
Generic ERL micro/nanoprobe camera
focus
Undulator source 5-25 keV rangeFocusing optics ~50 nm to ~5 m focus; automated switchingFresnel lens beam steering option for fast raster-scansCoherent beam optimize BL for coherent imagingRaster goniometer continuous scan capabilityPixel detector ≤50 m pixels;
Slide: 28XDL 2011 - 27&28 June- Christian Riekel