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Dive into the research topics where Hilton B. de Aguiar is active.

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Featured researches published by Hilton B. de Aguiar.


Optics Express | 2015

Quantitative analysis of light scattering in polarization-resolved nonlinear microscopy.

Hilton B. de Aguiar; Paulina Gasecka; Sophie Brasselet

Polarization resolved nonlinear microscopy (PRNM) is a powerful technique to gain microscopic structural information in biological media. However, deep imaging in a variety of biological specimens is hindered by light scattering phenomena, which not only degrades the image quality but also affects the polarization state purity. In order to quantify this phenomenon and give a framework for polarization resolved microscopy in thick scattering tissues, we develop a characterization methodology based on four wave mixing (FWM) process. More specifically, we take advantage of two unique features of FWM, meaning its ability to produce an intrinsic in-depth local coherent source and its capacity to quantify the presence of light depolarization in isotropic regions inside a sample. By exploring diverse experimental layouts in phantoms with different scattering properties, we study systematically the influence of scattering on the nonlinear excitation and emission processes. The results show that depolarization mechanisms for the nonlinearly generated photons are highly dependent on the scattering center size, the geometry used (epi/forward) and, most importantly, on the thickness of the sample. We show that the use of an un-analyzed detection makes the polarization-dependence read-out highly robust to scattering effects, even in regimes where imaging might be degraded. The effects are illustrated in polarization resolved imaging of myelin lipid organization in mouse spinal cords.


Physical Review A | 2016

Enhanced nonlinear imaging through scattering media using transmission matrix based wavefront shaping

Hilton B. de Aguiar; Sylvain Gigan; Sophie Brasselet

A method is proposed to optimize signals in nonlinear microscopy in order to circumvent difficulties associated with scattering media. In comparison to traditional nonlinear optimization techniques, the obtained results show a faster and more efficient imaging capability.


Science Advances | 2017

Polarization recovery through scattering media

Hilton B. de Aguiar; Sylvain Gigan; Sophie Brasselet

The lost polarization state purity of light is now shown to be recovered, after propagating in a strongly scattering environment. The control and use of light polarization in optical sciences and engineering are widespread. Despite remarkable developments in polarization-resolved imaging for life sciences, their transposition to strongly scattering media is currently not possible, because of the inherent depolarization effects arising from multiple scattering. We show an unprecedented phenomenon that opens new possibilities for polarization-resolved microscopy in strongly scattering media: polarization recovery via broadband wavefront shaping. We demonstrate focusing and recovery of the original injected polarization state without using any polarizing optics at the detection. To enable molecular-level structural imaging, an arbitrary rotation of the input polarization does not degrade the quality of the focus. We further exploit the robustness of polarization recovery for structural imaging of biological tissues through scattering media. We retrieve molecular-level organization information of collagen fibers by polarization-resolved second harmonic generation, a topic of wide interest for diagnosis in biomedical optics. Ultimately, the observation of this new phenomenon paves the way for extending current polarization-based methods to strongly scattering environments.Wavefront shaping has revolutionized imaging deep in scattering media, being able to spatially and temporally refocus light through or inside the medium. However, wavefront shaping is not compatible yet with polarization-resolved microscopy given the need of polarizing optics to refocus light with a controlled polarization state. Here, we show that wavefront shaping is not only able to restore a focus, but it can also recover the injected polarization state without using any polarizing optics at the detection. This counter-intuitive effect occurs up to several transport mean free path thick samples, which exhibit a speckle with a completely scrambled state. Remarkably, an arbitrary rotation of the input polarization does not degrade the quality of the focus. This unsupervised re-polarization - out of the originally scrambled polarization state - paves the way for polarization-resolved structural microscopy at unprecedented depths. We exploit this phenomenon and demonstrate second harmonic generation (SHG) structural imaging of collagen fibers in tendon tissues behind a scattering medium.


Optics Letters | 2017

Programmable single-pixel-based broadband stimulated Raman scattering

Pascal Berto; Camille Scotté; Frédéric Galland; Hervé Rigneault; Hilton B. de Aguiar

We report a simple add-on for broadband stimulated Raman scattering (SRS) microscopes to enable fast and programmable spectroscopy acquisition. It comprises a conventional dispersive spectrometer layout incorporating a fast digital micromirror device (DMD). The approach is validated by acquiring SRS spectra of standard chemicals. We demonstrate a DMDs advantage in broadband SRS by showing higher signal-to-noise ratio using a multiplexed Hadamard spectral basis and compressive sensing detection. Our results apply to a variety of frequency-domain pump-probe spectroscopy.


Journal of Physical Chemistry Letters | 2018

Molecular Imaging of Cholesterol and Lipid Distributions in Model Membranes

Stephen H. Donaldson; Hilton B. de Aguiar

Over recent decades, lipid membranes have become standard models for examining the biophysics and biochemistry of cell membranes. Interrogation of lipid domains within biomembranes is generally done with fluorescence microscopy via exogenous chemical probes. However, fluorophores have limited partitioning tunability, with the majority segregating into the liquid-disordered phase, and fluorescence only strictly reports on the small percentage of tagged lipids. We present simple, label-free imaging of domain formation in lipid monolayers, with chemical selectivity in unraveling lipid and cholesterol composition in different domain types. Exploiting conventional vibrational contrast in spontaneous Raman imaging, combined with chemometrics analysis, allows for examination of ternary systems containing saturated lipids, unsaturated lipids, and cholesterol. We confirm features commonly observed by fluorescence microscopy and provide a quantitative thermodynamic analysis of cholesterol distribution at the single-monolayer level.


Optica | 2017

Temporal recompression through a scattering medium via a broadband transmission matrix

Mickael Mounaix; Hilton B. de Aguiar; Sylvain Gigan

The transmission matrix is a unique tool to control light through a scattering medium. A monochromatic transmission matrix does not allow temporal control of broadband light. Conversely, measuring multiple transmission matrices with spectral resolution allows fine temporal control when a pulse is temporally broadened upon multiple scattering, but requires very long measurement time. Here, we show that a single linear operator, measured for a broadband pulse with a co-propagating reference, naturally allows for spatial focusing, and interestingly generates a two-fold temporal recompression at the focus, compared with the natural temporal broadening. This is particularly relevant for non-linear imaging techniques in biological tissues.


Journal of The Optical Society of America A-optics Image Science and Vision | 2018

Precision of proportion estimation with binary compressed Raman spectrum

Philippe Réfrégier; Camille Scotté; Hilton B. de Aguiar; Hervé Rigneault; Frédéric Galland

The precision of proportion estimation with binary filtering of a Raman spectrum mixture is analyzed when the number of binary filters is equal to the number of present species and when the measurements are corrupted with Poisson photon noise. It is shown that the Cramer-Rao bound provides a useful methodology to analyze the performance of such an approach, in particular when the binary filters are orthogonal. It is demonstrated that a simple linear mean square error estimation method is efficient (i.e., has a variance equal to the Cramer-Rao bound). Evolutions of the Cramer-Rao bound are analyzed when the measuring times are optimized or when the considered proportion for binary filter synthesis is not optimized. Two strategies for the appropriate choice of this considered proportion are also analyzed for the binary filter synthesis.


Analytical Chemistry | 2018

Assessment of Compressive Raman versus Hyperspectral Raman for Microcalcification Chemical Imaging

Camille Scotté; Hilton B. de Aguiar; Didier Marguet; Ellen Green; Pascaline Bouzy; Sébastien Vergnole; C.P. Winlove; Nicholas Stone; Hervé Rigneault

We experimentally implement a compressive Raman technology (CRT) that incorporates chemometric analysis directly into the spectrometer hardware by means of a digital micromirror device (DMD). The DMD is a programmable optical filter on which optimized binary filters are displayed. The latter are generated with an algorithm based on the Cramer-Rao lower bound. We compared the developed CRT microspectrometer with two conventional state-of-the-art Raman hyperspectral imaging systems on samples mimicking microcalcifications relevant for breast cancer diagnosis. The CRT limit of detection significantly improves, when compared to the CCD based system, and CRT ultimately allows 100× and 10× faster acquisition speeds than the CCD- and EMCCD-based systems, respectively.


Adaptive Optics and Wavefront Control for Biological Systems IV | 2018

Temporal recompression of an ultrashort pulse of light with a broadband transmission matrix (Conference Presentation)

Mickael Mounaix; Hilton B. de Aguiar; Sylvain Gigan

Spatial and temporal properties of an ultrashort pulse of light are naturally scrambled upon propagation in thick scattering media. Significant progresses have been realized over the last decade to manipulate light propagation in scattering media, mostly using monochromatic light. However, applications that require a broadband ultrashort pulse of light remain limited, as the pulse gets temporally broadened because of scattering effects. A monochromatic optical transmission matrix does not allow temporal control of broadband light. Although measuring multiple transmission matrices with spectral resolution allows fine temporal control, it requires lengthy measurements, as well as stability of the medium. In this work, we show that a single linear operator that we named Broadband Transmission Matrix, can be straightforwardly measured for a broadband pulse with a co-propagating reference. We exploit this operator for focusing purposes, and we analyze its phase conjugation properties. While the operator naturally allows for spatial focusing, unexpectedly, the focus duration is on average shorter than the natural temporal broadening due to the medium. More precisely, we observe a two-fold temporal recompression at the focus that we fully explain theoretically. We also explore the spectral content at the focus, and demonstrate a narrowing of the spectrum. These results are particularly relevant for non-linear imaging techniques in biological tissues, at depth where an ultrashort excitation pulse is broadened.


bioRxiv | 2017

Lipid order degradation in autoimmune demyelination probed by polarization resolved coherent Raman microscopy

Paulina Gasecka; Alexandre Jaouen; Fatma-Zohra Bioud; Hilton B. de Aguiar; Julien Duboisset; Patrick Ferrand; Hervé Rigneault; Naveen K. Balla; Franck Debarbieux; Sophie Brasselet

Myelin around axons is currently widely studied by structural analyses and large scale imaging techniques, with the goal to decipher its critical role in neuronal protection. While there is strong evidence that in myelin, lipid composition and lipid membrane morphology are affected during the progression of neurodegenerative diseases, there is no quantitative method yet to report its ultrastructure in tissues at both molecular and macroscopic levels, in conditions potentially compatible with in vivo observations. In this work, we study and quantify molecular order of lipids in myelin at sub-diffraction scales, using label-free polarization resolved Coherent Anti Stokes Raman (PR-CARS), which exploits CARS sensitivity to coupling between light polarization and oriented molecular vibrational bonds. Importantly, the method does not use any a priori parameters in the sample such as lipid type, orientational organization and composition. We show that lipid molecular order of myelin in the mouse spinal cord is significantly reduced throughout the progression of experimental autoimmune encephalomyelitis (EAE), a model for multiple sclerosis, even in myelin regions that appear morphologically unaffected. This technique permits to unravel molecular-scale perturbations of lipid layers at early stage of the demyelination progression, while the membrane architecture at the mesoscopic scale (here about 100 nm) seems much less affected. Such information cannot be brought by pure morphological observation and opens new prospectives towards molecular-scale understanding of neurodegenerative diseases.

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Hervé Rigneault

Université Paul Cézanne Aix-Marseille III

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