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Computational imaging with random encoding patterns obtained by scattering of light in complex media has enabled simple imaging systems with compelling performance. Here, we extend this concept to axial reflectivity profiling using spatio-temporal coupling of broadband light in a multimode fiber (MMF) to generate the encoding functions. Interference of light transmitted through the MMF with a sample beam results in path-length-specific patterns that enable computational reconstruction of the axial sample reflectivity profile from a single camera snapshot. Leveraging the versatile nature of MMFs, we demonstrate depth profiling with bandwidth-limited axial resolution of 13.4â µm over a scalable sensing range reaching well beyond one centimeter.
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Fossil teeth are primary tools in the study of vertebrate evolution, but standard imaging modalities have not been capable of providing high-quality images in dentin, the main component of teeth, owing to small refractive index differences in the fossilized dentin. Our first attempt to use third-harmonic generation (THG) microscopy in fossil teeth has yielded significant submicrometer level anatomy, with an unexpectedly strong signal contrasting fossilized tubules from the surrounding dentin. Comparison between fossilized and extant teeth of crocodilians reveals a consistent evolutionary signature through time, indicating the great significance of THG microscopy in the evolutionary studies of dental anatomy in fossil teeth.
Assuntos
Fósseis , Microscopia , Dente/anatomia & histologia , Jacarés e Crocodilos/anatomia & histologia , AnimaisRESUMO
In order to achieve high-resolution deep-tissue imaging, multi-photon fluorescence microscopy and photoacoustic tomography had been proposed in the past two decades. However, combining the advantages of these two imaging systems to achieve optical-spatial resolution with an ultrasonic-penetration depth is still a field with challenges. In this paper, we investigate the detection of the two-photon photoacoustic ultrasound, and first demonstrate background-free two-photon photoacoustic imaging in a phantom sample. To generate the background-free two-photon photoacoustic signals, we used a high-repetition rate femtosecond laser to induce narrowband excitation. Combining a loss modulation technique, we successfully created a beating on the light intensity, which not only provides pure sinusoidal modulation, but also ensures the spectrum sensitivity and frequency selectivity. By using the lock-in detection, the power dependency experiment validates our methodology to frequency-select the source of the nonlinearity. This ensures our capability of measuring the background-free two-photon photoacoustic waves by detecting the 2nd order beating signal directly. Furthermore, by mixing the nanoparticles and fluorescence dyes as contrast agents, the two-photon photoacoustic signal was found to be enhanced and detected. In the end, we demonstrate subsurface two-photon photoacoustic bio-imaging based on the optical scanning mechanism inside phantom samples.
Assuntos
Algoritmos , Aumento da Imagem/instrumentação , Aumento da Imagem/métodos , Microscopia Acústica/instrumentação , Microscopia Acústica/métodos , Microscopia de Fluorescência por Excitação Multifotônica/instrumentação , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Desenho de Equipamento , Análise de Falha de Equipamento , Dinâmica não LinearRESUMO
Dispersion remains an enduring challenge for the characterization of wavelength-dependent transmission through optical multimode fiber (MMF). Beyond a small spectral correlation width, a change in wavelength elicits a seemingly independent distribution of the transmitted field. Here we report on a parametric dispersion model that describes mode mixing in MMF as an exponential map and extends the concept of principal modes to describe the fiber's spectrally resolved transmission matrix (TM). We present computational methods to fit the model to measurements at only a few, judiciously selected, discrete wavelengths. We validate the model in various MMF and demonstrate an accurate estimation of the full TM across a broad spectral bandwidth, approaching the bandwidth of the best-performing principal modes, and exceeding the original spectral correlation width by more than two orders of magnitude. The model allows us to conveniently study the spectral behavior of principal modes, and obviates the need for dense spectral measurements, enabling highly efficient reconstruction of the multispectral TM of MMF.
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Imaging through optical multimode fiber (MMF) has the potential to enable hair-thin endoscopes that reduce the invasiveness of imaging deep inside tissues and organs. Active wavefront shaping and fluorescent labeling have recently been exploited to overcome modal scrambling and enable MMF imaging. Here, we present a computational approach that circumvents the need for active wavefront control and exogenous fluorophores. We demonstrate the reconstruction of depth-gated confocal images through MMF using a raster-scanned, focused input illumination at the fiber proximal end. We show the compatibility of this approach with quantitative phase, dark-field, and polarimetric imaging. Computational imaging through MMF opens a new pathway for minimally invasive imaging in medical diagnosis and biological investigations.
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Reciprocity is a fundamental principle of wave physics and directly relates to the symmetry in the transmission through a system when interchanging the input and output. The coherent transmission matrix (TM) is a convenient method to characterize wave transmission through general media. Here, we demonstrate the optical reciprocal nature of complex media by exploring their TM properties. We measured phase-corrected TMs of forward and round-trip propagation in a single polarization state through a looped 1 m-long step-index optical multimode fiber (MMF) to experimentally verify a transpose relationship between the forward and backward transmission. This symmetry impedes straightforward MMF calibration from proximal measurements of the round-trip TM. Furthermore, we show how focusing through the MMF with digital optical phase conjugation is compromised by system loss since time reversibility relies on power conservation. These insights may inform the development of new imaging techniques through complex media and coherent control of waves in photonic systems.
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Optical imaging is a key modality for observing biological specimen with higher spatial resolution. However, scattering and absorption of light in tissues are inherent barriers in maximizing imaging depth in biological tissues. To achieve this goal, use of light at near-infrared spectrum can improve the present situation. Here, the capability of saturated two-photon saturated excitation (TP-SAX) fluorescence microscopy to image at depths of >2.0 mm, with submicron resolution in transparent mouse brain imaging, is demonstrated. At such depths with scattering-enlarged point spread function (PSF), we find that TP-SAX is capable to provide spatial resolution improvement compared to its corresponding TPFM, which is on the other hand already providing a much improved resolution compared with single-photon confocal fluorescence microscopy. With the capability to further improve spatial resolution at such deep depth with scattering-enlarged PSF, TP-SAX can be used for exquisite visualization of delicate cerebral neural structure in the scattering regime with a submicron spatial resolution inside intact mouse brain.
Assuntos
Encéfalo/diagnóstico por imagem , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Rede Nervosa/diagnóstico por imagem , Animais , Encéfalo/metabolismo , Carbocianinas/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Rede Nervosa/metabolismo , Fenômenos ÓpticosRESUMO
Microscopy based on non-fluorescent absorption dye staining is widely used in various fields of biomedicine for 400 years. Unlike its fluorescent counterpart, non-fluorescent absorption microscopy lacks proper methodologies to realize its in vivo applications with a sub-femtoliter 3D resolution. Regardless of the most advanced high-resolution photoacoustic microscopy, sub-femtoliter spatial resolution is still unattainable, and the imaging speed is relatively slow. In this paper, based on the two-photon photoacoustic mechanism, we demonstrated a in vivo label free laser-scanning photoacoustic imaging modality featuring high frame rates and sub-femtoliter 3D resolution simultaneously, which stands as a perfect solution to 3D high resolution non-fluorescent absorption microscopy. Furthermore, we first demonstrated in vivo label-free two-photon acoustic microscopy on the observation of non-fluorescent melanin distribution within mouse skin.