An all-optical technique enables instantaneous single-shot demodulation of images at high frequency

Panigrahi, Swapnesh; Fade, Julien; Agaisse, Romain; Ramachandran, Hema; Alouini, Mehdi

doi:10.1038/s41467-019-14142-w

Cited by 14 publications

(10 citation statements)

References 34 publications

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“…Recently, innovative approaches toward a lock-in camera were proposed, based on advanced camera technology 138 or based on fast optical modulation of the polarization. 139 Lock-in cameras are based on the principle of each pixel demodulating the modulated signal similar to the lock-in amplifier. Therefore, lock-in cameras produce instantaneously amplitude and phase images.…”

Section: Mid-ir Photothermal Imaging Super-resolution Photothermal Imentioning

confidence: 99%

“…Therefore, lock-in cameras produce instantaneously amplitude and phase images. Panigrahi et al ( 139 ) proposed a polarization-based demodulation technique to obtain amplitude and phase information on a high-frequency (tens of kHz) modulated signal from a single acquisition of a camera image. The use of an electro-optic modulator and polarizing optical elements in a proper configuration creates four quadrature channels in a single recorded frame by an ordinary camera, and the careful image subtraction of these channels produces the final amplitude image.…”

Section: Mid-ir Photothermal Imaging Super-resolution Photothermal Imentioning

confidence: 99%

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Photothermal Microscopy: Imaging the Optical Absorption of Single Nanoparticles and Single Molecules

et al. 2020

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The photothermal (PT) signal arises from slight changes of the index of refraction in a sample due to absorption of a heating light beam. Refractive index changes are measured with a second probing beam, usually of a different color. In the past two decades, this all-optical detection method has reached the sensitivity of single particles and single molecules, which gave birth to original applications in material science and biology. PT microscopy enables shot-noise-limited detection of individual nanoabsorbers among strong scatterers and circumvents many of the limitations of fluorescence-based detection. This review describes the theoretical basis of PT microscopy, the methodological developments that improved its sensitivity toward single-nanoparticle and single-molecule imaging, and a vast number of applications to single-nanoparticle imaging and tracking in material science and in cellular biology.

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Section: Mid-ir Photothermal Imaging Super-resolution Photothermal Imentioning

confidence: 99%

Section: Mid-ir Photothermal Imaging Super-resolution Photothermal Imentioning

confidence: 99%

Photothermal Microscopy: Imaging the Optical Absorption of Single Nanoparticles and Single Molecules

et al. 2020

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“…In addition, the excitation signal must be switched off when the demodulation system spatially switches from a sub-region of the camera to another in order to avoid a spurious signal. We therefore propose another demodulation method based on polarization modulation using a Pockels cell enabling tunable high operating frequencies [17,26,27]. This implementation, shown in figure 6 a , requires that the fluorescent probes have sufficient degrees of freedom so that their emission is not polarized.…”

Section: Resultsmentioning

confidence: 99%

Time-modulated excitation for enhanced single-molecule localization microscopy

Jouchet

Poüs

Fort

et al. 2022

Phil. Trans. R. Soc. A.

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Structured illumination in single-molecule localization microscopy provides new information on the position of molecules and thus improves the localization precision compared to standard localization methods. Here, we used a time-shifted sinusoidal excitation pattern to modulate the fluorescence signal of the molecules whose position information is carried by the phase and recovered by synchronous demodulation. We designed two flexible fast demodulation systems located upstream of the camera, allowing us to overcome the limiting camera acquisition frequency and thus to maximize the collection of photons in the demodulation process. The temporally modulated fluorescence signal was then sampled synchronously on the same image, repeatedly during acquisition. This microscopy, called ModLoc, allows us to experimentally improve the localization precision by a factor of 2.4 in one direction, compared to classical Gaussian fitting methods. A temporal study and an experimental demonstration both show that the short lifetimes of the molecules in blinking regimes impose a modulation frequency in the kilohertz range, which is beyond the reach of current cameras. A demodulation system operating at these frequencies would thus be necessary to take full advantage of this new localization approach. This article is part of the Theo Murphy meeting issue 'Super-resolution structured illumination microscopy (part 2)'.

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“…22 Electrooptic imaging has since been applied to threedimensional modulation-enhanced localization microscopy, 23,24 lock-in profiling of optical modes, 25 and kilohertz imaging. 26 To enable practical applications in lifetime microscopy, improving the repetition rate while maintaining a usable field of view is essential. Here we use a resonant high-voltage drive technique to allow optical gating at 39 MHz.…”

Section: Introductionmentioning

confidence: 99%

Resonant Electro-optic Imaging for Microscopy at Nanosecond Resolution

Bowman,

Kasevich

2021

Preprint

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We present an electro-optic wide-field method to enable fluorescence lifetime microscopy (FLIM) with high throughput and single-molecule sensitivity. Resonantly driven Pockels cells are used to efficiently capture nanosecond image dynamics on standard camera sensors. Images are gated at 39 MHz with 16 dB extinction and 86% modulation depth, allowing lifetimes to be estimated with sensitivity 1.9 times the shot-noise limit and without restricting photon throughput. High throughput is shown by acquiring FLIM images with millisecond exposure and > 10 8 photons per image. High sensitivity is shown through wide-field lifetime imaging of single-molecule populations and dynamics. Using both the throughput and sensitivity of the technique, we enable the first combination of wide-field lifetime imaging with single-molecule localization microscopy. We further demonstrate lifetime imaging of single-molecule FRET and multi-parameter imaging of molecular binding events combining intensity, lifetime, and polarization. More generally, resonant electro-optic imaging can provide lifetime contrast in any wide-field method and will enable nanosecond time-domain measurements in microscopy.

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An all-optical technique enables instantaneous single-shot demodulation of images at high frequency

Cited by 14 publications

References 34 publications

Photothermal Microscopy: Imaging the Optical Absorption of Single Nanoparticles and Single Molecules

Photothermal Microscopy: Imaging the Optical Absorption of Single Nanoparticles and Single Molecules

Time-modulated excitation for enhanced single-molecule localization microscopy

Resonant Electro-optic Imaging for Microscopy at Nanosecond Resolution

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