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1.
Biochemistry (Mosc) ; 84(Suppl 1): S124-S143, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-31213199

RESUMO

Diabetes mellitus is a metabolic disorder characterized by chronic hyperglycemia accompanied by the disruption of carbohydrate, lipid, and proteins metabolism and development of long-term microvascular, macrovascular, and neuropathic changes. This review presents the results of spectroscopic studies on the glycation of tissues and cell proteins in organisms with naturally developing and model diabetes and in vitro glycated samples in a wide range of electromagnetic waves, from visible light to terahertz radiation. Experiments on the refractometric measurements of glycated and oxygenated hemoglobin in broad wavelength and temperature ranges using digital holographic microscopy and diffraction tomography are discussed, as well as possible application of these methods in the diabetes diagnostics. It is shown that the development and implementation of multimodal approaches based on a combination of phase diagnostics with other methods is another promising direction in the diabetes diagnostics. The possibilities of using optical clearing agents for monitoring the diffusion of substances in the glycated tissues and blood flow dynamics in the pancreas of animals with induced diabetes have also been analyzed.


Assuntos
Diabetes Mellitus/diagnóstico por imagem , Hemoglobina A Glicada/ultraestrutura , Animais , Velocidade do Fluxo Sanguíneo , Glicosilação , Holografia/métodos , Humanos , Microscopia/métodos , Espectrofotometria Infravermelho/métodos , Espectroscopia Terahertz/métodos , Tomografia/métodos
2.
Appl Opt ; 58(15): 4042-4046, 2019 May 20.
Artigo em Inglês | MEDLINE | ID: mdl-31158157

RESUMO

This paper utilized digital holographic microscopy and optical tweezers to study microdeformation of red blood cells (RBCs) dynamically under oxidative stress. RBCs attached with microbeads were stretched by dual optical tweezers to generate microdeformation. Morphology of RBCs under manipulation were recorded dynamically and recovered by off-axis digital holographic microscopy method. RBCs treated with H2O2 at different concentrations were measured to investigate the mechanical properties under oxidative stress. Use of optical tweezers and off-axis digital holographic microscopy enhanced measuring accuracy compared with the traditional method. Microdeformation of RBCs is also more consistent with the physiological situation. This proposal is meaningful for clinical applications and basic analysis of Parkinson's disease research.


Assuntos
Deformação Eritrocítica/fisiologia , Eritrócitos/citologia , Holografia/métodos , Pinças Ópticas , Estresse Oxidativo , Forma Celular/fisiologia , Elasticidade/fisiologia , Humanos , Microscopia/métodos
3.
Opt Express ; 27(10): 13581-13595, 2019 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-31163820

RESUMO

Lens-free holographic microscopy (LFHM) provides a cost-effective tool for large field-of-view imaging in various biomedical applications. However, due to the unit optical magnification, its spatial resolution is limited by the pixel size of the imager. Pixel super-resolution (PSR) technique tackles this problem by using a series of sub-pixel shifted low-resolution (LR) lens-free holograms to form the high-resolution (HR) hologram. Conventional iterative PSR methods require a large number of measurements and a time-consuming reconstruction process, limiting the throughput of LFHM in practice. Here we report a deep learning-based PSR approach to enhance the resolution of LFHM. Compared with the existing PSR methods, our neural network-based approach outputs the HR hologram in an end-to-end fashion and maintains consistency in resolution improvement with a reduced number of LR holograms. Moreover, by exploiting the resolution degradation model in the imaging process, the network can be trained with a data set synthesized from the LR hologram itself without resorting to the HR ground truth. We validated the effectiveness and the robustness of our method by imaging various types of samples using a single network trained on an entirely different data set. This deep learning-based PSR approach can significantly accelerate both the data acquisition and the HR hologram reconstruction processes, therefore providing a practical solution to fast, lens-free, super-resolution imaging.


Assuntos
Holografia/métodos , Aumento da Imagem/métodos , Microscopia/métodos , Algoritmos , Aprendizado de Máquina
4.
Opt Express ; 27(10): 13706-13720, 2019 May 13.
Artigo em Inglês | MEDLINE | ID: mdl-31163830

RESUMO

An outstanding challenge for immunology is the classification of immune cells in a label-free fashion with high speed. For this purpose, optical techniques such as Raman spectroscopy or digital holographic microscopy have been used successfully to identify immune cell subsets. To achieve high accuracy, these techniques require a post-processing step using linear methods of multivariate processing, such as principal component analysis. Here we demonstrate for the first time a comparison between artificial neural networks and principal component analysis (PCA) to classify the key granulocyte cell lineages of neutrophils and eosinophils using both digital holographic microscopy and Raman spectroscopy. Artificial neural networks can offer advantages in terms of classification accuracy and speed over a PCA approach. We conclude that digital holographic microscopy with convolutional neural networks based analysis provides a route to a robust, stand-alone and high-throughput hemogram with a classification accuracy of 91.3 % at a throughput rate of greater than 100 cells per second.


Assuntos
Eosinófilos/citologia , Holografia/métodos , Neutrófilos/citologia , Análise Espectral Raman/métodos , Linhagem da Célula , Separação Celular/métodos , Citometria de Fluxo , Humanos , Análise de Componente Principal
5.
J Clin Neurosci ; 67: 234-238, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31221576

RESUMO

During neurological surgery, neurosurgeons have to transform the two-dimensional (2D) sectional images into three-dimensional (3D) structures at the cognitive level. The complexity of the intracranial structures increases the difficulty and risk of neurosurgery. Mixed reality (MR) applications reduce the obstacles in the transformation from 2D images to 3D visualization of anatomical structures of central nervous system. In this study, the holographic image was established by MR using computed tomography (CT), computed tomography angiography (CTA) and magnetic resonance imaging (MRI) data of patients. The surgeon's field of vision was superimposed with the 3D model of the patient's intracranial structure displayed on the mixed reality head-mounted display (MR-HMD). The neurosurgeons practiced and evaluated the feasibility of this technique in neurosurgical cases. We developed the segmentation image masks and texture mapping including brain tissue, intracranial vessels, nerves, tumors, and their relative positions by MR technologies. The results showed that the three-dimensional imaging is in a stable state in the operating room with no significant flutter and blur. And the neurosurgeon's feedback on the comfort of the equipment and the practicality of the technology was satisfactory. In conclusion, MR technology can holographically construct a 3D digital model of patient's lesions and improve the anatomical perception of neurosurgeons during craniotomy. The feasibility of the MR-HMD application in neurosurgery is confirmed.


Assuntos
Craniotomia/métodos , Holografia/métodos , Cirurgia Assistida por Computador/métodos , Idoso , Neoplasias Encefálicas/cirurgia , Feminino , Humanos , Imagem por Ressonância Magnética/métodos , Masculino , Pessoa de Meia-Idade , Tomografia Computadorizada por Raios X/métodos
6.
Appl Opt ; 58(12): 3104-3114, 2019 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-31044784

RESUMO

Modern microscopes are designed with functionalities that are tailored to enhance image contrast. Dark-field imaging, phase contrast, differential interference contrast, and other optical techniques enable biological cells and other phase-only objects to be visualized. Quantitative phase imaging refers to an emerging set of techniques that allow for the complex transmission function of the sample to be measured. With this quantitative phase image available, any optical technique can then be simulated; it is trivial to generate a phase contrast image or a differential interference contrast image. Rheinberg illumination, proposed almost a century ago, is an optical technique that applies color contrast to images of phase-only objects by introducing a type of optical staining via an amplitude filter placed in the illumination path that consists of two or more colors. In this paper, the complete theory of Rheinberg illumination is derived, from which an algorithm is proposed that can digitally simulate the technique. Results are shown for a number of quantitative phase images of diatom cells obtained via digital holographic microscopy. The results clearly demonstrate the potential of the technique for label-free color staining of subcellular features.


Assuntos
Diatomáceas/citologia , Holografia/métodos , Iluminação , Microscopia de Contraste de Fase/métodos , Coloração e Rotulagem/métodos , Algoritmos
7.
Appl Opt ; 58(5): A258-A266, 2019 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-30873999

RESUMO

The computer-generated hologram (CGH) technique is a technique that simulates the recording of holography. Although the CGH technique has a lot of advantages, it also has some disadvantages; one of them is the long calculation time. Much research on the human eye has established that humans see 135° vertically and 160° horizontally, but can see fine detail within an only 5° central circle. Foveated rendering uses this characteristic of the human eye to reduce image resolution in the peripheral area and achieve a high calculation speed. In this paper, a new method for CGH fast calculation with foveated rendering using an angle-changeable ray-tracing method is introduced. The experiments demonstrate the effectiveness and high-speed calculation of this method.


Assuntos
Holografia/métodos , Aumento da Imagem/métodos , Algoritmos , Desenho de Equipamento , Humanos , Imagem Tridimensional/métodos , Processamento de Sinais Assistido por Computador , Acuidade Visual/fisiologia
8.
Molecules ; 24(6)2019 Mar 14.
Artigo em Inglês | MEDLINE | ID: mdl-30875723

RESUMO

Holographic volume phase gratings are recorded in an epoxy-based, free-surface, volume holographic recording material. Light-induced gratings are formed by photo-triggered mass migration caused by component diffusion. The material resolution enables a wide range of pattern spacings, to record both transmission and reflection holograms with many different spatial frequencies. An optimum spatial frequency response is found between the low spatial frequency roll-off and the high spatial frequency cut-off. The influence of the energy density of exposure on the spatial frequency response is investigated. Secondary volume holographic gratings (parasitic gratings) are observed in the high frequency range. The possibility of distinguishing the regular grating from the secondary grating is discussed in the form of probe wavelength detuning.


Assuntos
Holografia/métodos , Fármacos Fotossensibilizantes/química , Polímeros/química , Luz , Refratometria , Propriedades de Superfície
9.
Sensors (Basel) ; 19(5)2019 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-30862101

RESUMO

Continuous cell culture monitoring as a way of investigating growth, proliferation, and kinetics of biological experiments is in high demand. However, commercially available solutions are typically expensive and large in size. Digital inline-holographic microscopes (DIHM) can provide a cost-effective alternative to conventional microscopes, bridging the gap towards live-cell culture imaging. In this work, a DIHM is built from inexpensive components and applied to different cell cultures. The images are reconstructed by computational methods and the data are analyzed with particle detection and tracking methods. Counting of cells as well as movement tracking of living cells is demonstrated, showing the feasibility of using a field-portable DIHM for basic cell culture investigation and bringing about the potential to deeply understand cell motility.


Assuntos
Rastreamento de Células/métodos , Microscopia/métodos , Técnicas de Cultura de Células , Holografia/métodos , Humanos
10.
Opt Lett ; 44(4): 995-998, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30768044

RESUMO

In this Letter, a concept of new multi-parameter imaging that can acquire visual and audio data of dynamic object phenomena simultaneously by a holographic technique is proposed. Temporal intensity distributions give us visual information of the dynamic events. The temporal profile of the phase distribution can give different information of the dynamic events, such as audio data. These two physical data can express the dynamic events with multi-parameters in various dimensions. The proposed imaging approach has potential in several applications in physics and life science research. Two experimental demonstrations such as static object and broken glass with visual and sound features are provided to show the effectiveness of the proposed concept.


Assuntos
Holografia/métodos , Som , Visão Ocular , Humanos
11.
Nat Protoc ; 14(3): 864-900, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30804570

RESUMO

Optogenetic tools provide users the ability to photocontrol the activity of cells. Commonly, activation is achieved by expression of proteins from photosynthetic organisms, for example, microbial opsins (e.g., ChR2). Alternatively, a sister approach, synthetic optogenetics, enables photocontrol over proteins of mammalian origin by use of photoswitches, visible light (typically), and genetic modification. Thus, synthetic optogenetics facilitates interrogation of native neuronal signaling mechanisms. However, the poor tissue penetration of visible wavelengths impedes the use of the technique in tissue, as two-photon excitation (2PE) is typically required to access the near-infrared window. Here, we describe an alternative technique that uses 2PE-compatible photoswitches (section 1) for photoactivation of genetically modified glutamate receptors (section 2). Furthermore, for fast, multi-region photoactivation, we describe the use of 2P-digital holography (2P-DH) (section 3). We detail how to combine 2P-DH and synthetic optogenetics with electrophysiology, or with red fluorescence Ca2+ recordings, for all-optical neural interrogation. The time required to complete the methods, aside from obtaining the necessary reagents and illumination equipment, is ~3 weeks.


Assuntos
Holografia/métodos , Optogenética/métodos , Fótons , Sequência de Aminoácidos , Animais , Compostos Azo/química , Feminino , Células HEK293 , Humanos , Ligantes , Domínios Proteicos , Ratos Sprague-Dawley , Receptores de Glutamato/química , Estereoisomerismo
12.
Analyst ; 144(5): 1751-1760, 2019 Feb 25.
Artigo em Inglês | MEDLINE | ID: mdl-30666996

RESUMO

The accurate and fast size classification of microparticles is important in environmental monitoring and biomedical applications. Conventional methods for sensing and classifying microparticles require bulky optical setups and generally show medium performance. Accordingly, the development of a portable and smart platform for accurate particle size classification is essential. In this study, we propose a new sensing platform for automatic identification of microparticle types through the synergistic integration of smartphone-based digital in-line holographic microscopy (DIHM) and machine-learning algorithms. The smartphone-based DIHM system consists of a coherent laser beam, a pinhole, a sample holder, a three-dimensional printed attachment, and a modified built-in smartphone camera module. The portable device has a physical dimension of 4 × 8 × 10 cm3 and 220 g in weight. Holograms of various polystyrene microparticles with different sizes (d = 2-50 µm) were recorded with a wide field-of-view and high spatial resolution. To establish a proper classification model, tens of features including geometrical parameters and light-intensity distributions were extracted from holograms of individual particles, and five machine-learning algorithms were used. After examining the performance of several classifiers, the resulting support vector machine model trained by using three geometrical parameters and three extracted parameters from light-intensity distributions shows the highest accuracy in the particle classification of the training and test sets (>98%). Therefore, the developed handheld smartphone-based platform can be potentially utilized to cope with various imaging needs in mobile healthcare and environmental monitoring.


Assuntos
Holografia/instrumentação , Microscopia/instrumentação , Tamanho da Partícula , Poliestirenos/química , Poliestirenos/classificação , Smartphone , Algoritmos , Holografia/métodos , Aprendizado de Máquina , Microscopia/métodos
13.
PLoS One ; 14(1): e0210340, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30677070

RESUMO

We propose the fabrication of surface relief holograms via selective SiO2 deposition on soda-lime silicate glass substrates. Initially, the original hologram was recorded on an azobenzene photosensitive polymer film coated on the soda-lime silicate glass by irradiation with a conventional continuous wave Ar+ laser with a wavelength of 514.5 nm. The hologram was transferred to the soda-lime silicate glass surface via a corona discharge treatment as an index modulation hologram, which was created by partial substitution of protons for sodium ions during the corona discharge treatment in air. After the corona discharge treatment, the polymer film was removed from the substrate. The diffraction efficiency of the index hologram on the soda-lime silicate glass was estimated to be 5.8 × 10-2% at a wavelength of 532 nm. Finally, the glass substrate was subjected to corona discharge treatment in air with vaporized cyclic siloxane. A surface relief hologram with the diffraction efficiency of 2.3% was successfully fabricated on the soda-lime silicate glass.


Assuntos
Holografia/métodos , Compostos de Cálcio , Vidro , Lasers de Gás , Microscopia de Força Atômica , Óxidos , Processos Fotoquímicos , Silicatos , Dióxido de Silício , Hidróxido de Sódio , Propriedades de Superfície
14.
PLoS One ; 14(1): e0210564, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30689635

RESUMO

Patterned two-photon (2P) photolysis via holographic illumination is a powerful method to investigate neuronal function because of its capability to emulate multiple synaptic inputs in three dimensions (3D) simultaneously. However, like any optical system, holographic projectors have a finite space-bandwidth product that restricts the spatial range of patterned illumination or field-of-view (FOV) for a desired resolution. Such trade-off between holographic FOV and resolution restricts the coverage within a limited domain of the neuron's dendritic tree to perform highly resolved patterned 2P photolysis on individual spines. Here, we integrate a holographic projector into a commercial 2P galvanometer-based 2D scanning microscope with an uncaging unit and extend the accessible holographic FOV by using the galvanometer scanning mirrors to reposition the holographic FOV arbitrarily across the imaging FOV. The projector system utilizes the microscope's built-in imaging functions. Stimulation positions can be selected from within an acquired 3D image stack (the volume-of-interest, VOI) and the holographic projector then generates 3D illumination patterns with multiple uncaging foci. The imaging FOV of our system is 800×800 µm2 within which a holographic VOI of 70×70×70 µm3 can be chosen at arbitrary positions and also moved during experiments without moving the sample. We describe the design and alignment protocol as well as the custom software plugin that controls the 3D positioning of stimulation sites. We demonstrate the neurobiological application of the system by simultaneously uncaging glutamate at multiple spines within dendritic domains and consequently observing summation of postsynaptic potentials at the soma, eventually resulting in action potentials. At the same time, it is possible to perform two-photon Ca2+ imaging in 2D in the dendrite and thus to monitor synaptic Ca2+ entry in selected spines and also local regenerative events such as dendritic action potentials.


Assuntos
Holografia/métodos , Imagem Tridimensional , Estimulação Luminosa , Fótons , Animais , Ratos Wistar , Software , Sinapses/fisiologia
15.
J Cardiothorac Vasc Anesth ; 33(3): 732-741, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30340952

RESUMO

OBJECTIVE: The use of 3-dimensional (3D) transesophageal echocardiography (TEE) in perioperative evaluation of the mitral valve (MV) is increasing progressively, including the use of 3D MV models for quantitative analysis. However, the use of 3D MV models in clinical practice still is limited by the need for specific training and the long time required for analysis. A new stereoscopic visualization tool (EchoPixel True 3D) allows virtual examination of anatomic structures in the clinical setting, but its accuracy and feasibility for intraoperative use is unknown. The aim of this study was to assess the feasibility of 3D holographic display and evaluate 3D quantitative measurements on a volumetric MV image using the EchoPixel system compared with the 3D MV model generated by QLAB Mitral Valve Navigation (MVN) software. DESIGN: This was a retrospective comparative study. SETTING: The study took place in a tertiary care center. PARTICIPANTS: A total of 40 patients, 20 with severe mitral regurgitation who underwent mitral valve repair and 20 controls with normal MV, were enrolled retrospectively. INTERVENTIONS: The 3D-TEE datasets of the MV were analyzed using a 3D MV model and stereoscopic display. The agreement of measurements, intraobserver and interobserver variability, and time for analysis were assessed. MEASUREMENTS AND MAIN RESULTS: Fair agreement between the 2 software systems was found for annular circumference and area in pathologic valves, but good agreement was reported for prolapse height and linear annular diameters. A higher agreement for all annular parameters and prolapse height was seen in normal valves. Excellent intraobserver and interobserver reliability was proved for the same parameters; time for analysis between the 2 methods in pathologic valves was substantially equivalent, although longer in pathologic valves when compared with normal MV using both tools. CONCLUSION: EchoPixel proved to be reliable to display 3D TEE datasets and accurate for direct linear measurement of both MV annular sizes and prolapse height compared to QLAB MVN software; it also carries a low interobserver and intraobserver variability for most measurements.


Assuntos
Ecocardiografia Tridimensional/normas , Ecocardiografia Transesofagiana/normas , Holografia/normas , Insuficiência da Valva Mitral/diagnóstico por imagem , Idoso , Ecocardiografia Tridimensional/métodos , Ecocardiografia Transesofagiana/métodos , Feminino , Holografia/métodos , Humanos , Masculino , Pessoa de Meia-Idade , Insuficiência da Valva Mitral/fisiopatologia , Variações Dependentes do Observador , Reprodutibilidade dos Testes , Estudos Retrospectivos
16.
Sci Rep ; 8(1): 17003, 2018 11 19.
Artigo em Inglês | MEDLINE | ID: mdl-30451953

RESUMO

Lens-free digital in-line holography (LDIH) is a promising microscopic tool that overcomes several drawbacks (e.g., limited field of view) of traditional lens-based microcopy. However, extensive computation is required to reconstruct object images from the complex diffraction patterns produced by LDIH. This limits LDIH utility for point-of-care applications, particularly in resource limited settings. We describe a deep transfer learning (DTL) based approach to process LDIH images in the context of cellular analyses. Specifically, we captured holograms of cells labeled with molecular-specific microbeads and trained neural networks to classify these holograms without reconstruction. Using raw holograms as input, the trained networks were able to classify individual cells according to the number of cell-bound microbeads. The DTL-based approach including a VGG19 pretrained network showed robust performance with experimental data. Combined with the developed DTL approach, LDIH could be realized as a low-cost, portable tool for point-of-care diagnostics.


Assuntos
Algoritmos , Aprendizado Profundo , Holografia/métodos , Processamento de Imagem Assistida por Computador/métodos , Neoplasias/classificação , Neoplasias/diagnóstico , Biomarcadores Tumorais/metabolismo , Humanos , Aumento da Imagem , Aprendizado de Máquina , Neoplasias/metabolismo , Patologia Molecular , Células Tumorais Cultivadas
17.
Opt Express ; 26(20): 26520-26533, 2018 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-30469737

RESUMO

For the see-through and near-to-eye displays, light throughput and uniformity of luminance over the field of view are improved by employing an optical image guide with discretely depth-varying surface relief holographic gratings. In the design process, a newly developed mathematical model, in conjunction with rigorous coupled wave analysis of diffraction efficiency, eliminates massive and time consuming iteration of non-sequential ray tracing but rapidly identifies the depth-varying structure and optimum optical performance. The depth-varying grating based approach achieved a 1.37x improvement in light throughput compared to the conventional depth un-varying design, 315 cd/m2/lm, along with improved uniformity over the field of view of 35 (H) x 20 (V) degrees with an eye box size of 17 (H) x 14 (V) mm.


Assuntos
Simulação por Computador , Holografia/instrumentação , Modelos Teóricos , Algoritmos , Desenho de Equipamento , Holografia/métodos , Processamento de Imagem Assistida por Computador , Refratometria/métodos
18.
Opt Express ; 26(21): 27076-27088, 2018 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-30469782

RESUMO

We propose an optical see-through holographic near-eye-display that can control the depth of field of individual virtual three-dimensional image and replicate the eyebox with dynamic steering. For optical see-through capability and eyebox duplication, a holographic optical element is used as an optical combiner where it functions as multiplexed tilted concave mirrors forming multiple copies of the eyebox. For depth of field control and eyebox steering, computer generated holograms of three-dimensional objects are synthesized with different ranges of angular spectrum. In optical experiment, it has been confirmed that the proposed system can present always-focused images with large depth of field and three-dimensional images at different distances with shallow depth of field at the same time without any time-multiplexing.


Assuntos
Algoritmos , Holografia/métodos , Imagem Tridimensional , Dispositivos Ópticos , Simulação por Computador
19.
Microsc Res Tech ; 81(12): 1361-1365, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30431202

RESUMO

A new blood flow imaging (BFI) technique using digital holography with double illumination of the sample is proposed. We imaged the moving red blood cells (RBCs) using a two microscope objective lenses setup. The setup consists in a larger angle of separation (90 °) between the two illumination beams, allowing a wider angular rotation at good z resolution. Moreover, the setup geometry allows an easier displacement of the sample in all directions. Results show that this technique is able to perform phase-shifting reconstruction for the two beams at the same time which is more suitable for the future implementation of live 3D holography. Experimental results are carried out for the verification of the effectiveness of the proposed technique on a zebrafish larvae sample. RESEARCH HIGHLIGHTS: Blood flow imaging techniques are often invasive and image analysis is time consuming. To alleviate this issue an imaging technique based on dual illumination in holographic domain is proposed. This method has been validated on zebrafish embryos.


Assuntos
Holografia/métodos , Imagem Tridimensional/métodos , Microscopia/métodos , Peixe-Zebra/sangue , Animais , Desenho de Equipamento , Eritrócitos/química , Eritrócitos/citologia , Larva/química , Larva/fisiologia , Peixe-Zebra/embriologia , Peixe-Zebra/fisiologia
20.
J Biomed Opt ; 23(11): 1-8, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30444085

RESUMO

Conventional temporal focusing-based multiphoton excitation microscopy (TFMPEM) can offer widefield optical sectioning with an axial excitation confinement of a few microns. To improve the axial confinement of TFMPEM, a binary computer-generated Fourier hologram (CGFH) via a digital-micromirror-device (DMD) was implemented to intrinsically improve the axial confinement by filling the back-focal aperture of the objective lens. Experimental results show that the excitation focal volume can be condensed and the axial confinement improved about 24% according to the DMD holography. In addition, pseudouniform MPE can be achieved using two complementary CGFHs with rapid pulse-width modulation switching via the DMD. Furthermore, bioimaging of CV-1 in origin with SV40 genes-7 cells demonstrates that the TFMPEM with binary DMD holography can improve image quality by enhancing axial excitation confinement and rejecting out-of-focus excitation.


Assuntos
Holografia/métodos , Processamento de Imagem Assistida por Computador/métodos , Microscopia de Fluorescência por Excitação Multifotônica/métodos , Animais , Células COS , Desenho de Equipamento , Holografia/instrumentação , Lasers , Microscopia de Fluorescência por Excitação Multifotônica/instrumentação
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