Quantum Super-resolution Imaging in Fluorescence Microscopy
Dept. of Physics of Complex SystemsWeizmann Institute of Science, Israel
FRISNO 12, Ein Gedi (February 2013)
Osip Schwartz, Dan Oron, Jonathan M. Levitt, Ron Tenne, Stella Itzhakov and Dan Oron
Slide 2 of 18
Microscopy and resolution
Workarounds:•Nonlinear optical methods: use nonlinear optical response to produce narrower point spread function
•Stochastic methods: use fluorophores turning on and off randomly
•Quantum optics?
Resolution of far-field optical microscopes is limited by about half wavelength.(Ernst Abbe, 1873)
o Multi-photon interference Afek et al., Science 328 (2010) Walther et al., Nature 429 (2004)
o Entangled images Boyer et al., Science 321 (2008)o Sub shot noise imaging Brida et al., Nat. Photonics 4 (2010)o Resolution enhancement?
Slide 3 of 18
Object Light detecto
r
Imaging system
Quantum light
Quantum super-resolution
Kolobov, Fabre, PRL2000
Saleh et al., PRL 2005
M.Tsang PRL 2009
Shin et al., PRL 2011
Thiel et al., PRL 2007
Thiel et al., PRA 2009
Giovannetti, PRA 2009
Guerrieri et al., PRL 2010
• Quantum Limits on Optical Resolution
• Wolf equations for two-photon light
• Quantum Imaging beyond the Diffraction Limit by Optical Centroid Measurements
• Quantum spatial superresolution by optical centroid measurements
• Quantum imaging with incoherent photons,
• Sub-Rayleigh quantum imaging using single-photon sources
• Sub-Rayleigh-diffraction-bound quantum imaging,
• Sub-Rayleigh Imaging via N-Photon Detection,
Slide 4 of 18
Classical light
Quantum emitters
Light detector
Imaging system
Quantum light
Quantum emitters
S.W. Hell et al., Bioimaging (1995)
What if we had an emitter that would always emit photon pairs?
Slide 5 of 18
Multi-photon detection microscopy
Spatial distribution of photon pairs carries high spatial frequency information (up to double resolution)
cascaded emitters
Photon pair detector
Imaging system
Photon pairτ1
τ2
τ1>>τ2
Point spread function: h2phot(x) = h2(x)
Similarly, in N-photon detection microscopy hNphot(x) = hN(x)
Slide 6 of 18
Antibunching microscopy
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Instead of actual photon pairs, consider ‘missing’ pairs.
Fluorescence intensity autocorrelation g(2)
10 μs interval between pulses
Number of photons emitted after excitation:Organic dyes: W. Ambrose et al. (1997)
Quantum dots: B. Lounis et al. (2000).
NV centers: R. Brouri et al. (2000).
Observations of antibunching:
Slide 7 of 18
Antibunching-induced correlations
Two adjacent detectors in the image plane:
x0
For individual fluorophore:For individual fluorophore:
Sum over fluorophores
For multiple fluorophores:For multiple fluorophores:
Slide 8 of 18
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Wavelength, nm
EmittersFluorescence saturation
Schwartz et al.,ACS Nano 6 (2012)
CdSe / ZnSe / ZnS quantum dots
Slide 9 of 18
At 1 kHz:
Schwartz et al.,ACS Nano 6 (2012)
Slide 10 of 18
Photon counting with a CCD
threshold
Dark counts
Less noiseMore signal
Read noisePixel signal distribution
CCD ADC units
arXiv:1212.6003
Slide 11 of 18
Computing correlations
• compute correlations for all pixel configurations
• Fourier-interpolate the resulting images
• Sum the interpolated images
2nd order:
3rd order:
Quantifies the missing pairs
Missing 3-photon events (except those due to missing pairs, already accounted for)
arXiv:1212.6003
Slide 12 of 18
Antibunching with a CCD
Third order:g(3)(τ1, τ2)==<n(t)n(t+τ1)n(t+ τ2)>
Second order autocorrelation function:g(2)(τ)=<n(t)n(t+ τ)>
τ, ms τ, ms
τ1, ms τ1, ms
τ 2, m
sτ 2,
ms
Quantum dot Classical signal
arXiv:1212.6003
Slide 13 of 18 arXiv:1212.6003
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Fluorescence image
Slide 14 of 18 arXiv:1212.6003
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Fluorescence image
2nd order antibunching
Slide 15 of 18 arXiv:1212.6003
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Fluorescence image
2nd order antibunching
3rd order antibunching
Resolution: 271 nm FWHM
216 nm FWHM(x1.26)
181 nm FWHM(x1.50)
Slide 16 of 18
Optical sectioning
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Fluorescence imaging
2nd order antibunching
imaging
Defocused image of a quantum dot:
Optical signal integrated over the field of view:
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Defocusing, μm
Slide 17 of 18
Summary
•Far-field super-resolution imaging demonstrated by using quantum properties of light naturally present in fluorescence microscopy
•The experiment was performed with commercially available equipment, at room temperature, with commonly used quantum dot fluorophores
•With further development of detector technology, antibunching imaging may become feasible as a practical imaging method
Slide 18 of 18
The team
Jonathan M. Levitt
Dan Oron
Zvicka DeutschStella Itzhakov
Ron Tenne
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Superresolved images
Regular (photon counting) image Second order correlations Third order correlations
Reconstructed high resolution images
Slide 21 of 18
Superresolved images
arXiv:1212.6003
Slide 22 of 18
Superresolved images
Slide 23 of 18
Superresolved images
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PSF width (nm)
PSF width (nm)
PSF width (nm)
Slide 25 of 18
Quantum super-resolution
Conceptual difficulty: an absorptive grating with sub-wavelength period acts as an attenuator for every photon
• Transmitted light contains no information on the grating phase or period • Any linear absorber mask is a superposition of gratings• High spatial frequency components of the mask are lost
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