Terahertz interferometric synthetic aperture tomography for confocal imaging systems

Terahertz (THz) interferometric synthetic aperture tomography (TISAT) for confocal imaging within extended objects is demonstrated by combining attributes of synthetic aperture radar and optical coherence tomography. Algorithms recently devised for interferometric synthetic aperture microscopy are adapted to account for the diffraction-and defocusing-induced spatially varying THz beam width characteristic of narrow depth of focus, high-resolution confocal imaging. A frequency-swept two-dimensional TISAT confocal imaging instrument rapidly achieves in-focus, diffraction-limited resolution over a depth 12 times larger than the instrument's depth of focus in a manner that may be easily extended to three dimensions and greater depths.     

Three-dimensional integral imaging of micro-objects

We propose a method for displaying micro-objects in space that is based on three-dimensional (3D) integral imaging, in which elemental images are calculated from a two-dimensional sampling of the optical field along different depths by use of confocal scanning microscopy. Experimental results are presented to demonstrate that a uniformly magnified 3D biological specimen can be displayed in space, and thus integral imaging can be used for 3D display of confocal microscopy. To the best of our knowledge, this is the first report of 3D integral imaging of (semitransparent) micro-objects.     

基于深度变化成像模型的调整EM算法

在实际成像中,通常样本中的物质是变化的,故样本中不同位置的折射率不一样。由于三维样本的折射率与物镜所浸物质的折射率的不匹配,导致不同深度的点扩展函数可能不同。在此深度变化成像模型基础上应用最大期望(EM)复原算法能够提高图像清晰度,尤其是深度方向,但会丢失图像的一些微弱细节且出现一些孤立亮点,因此将调整EM算法运用到基于三维显微光学切片中成像随深度变化的图像模型上,此二者结合后的新算法可以避免上述缺点,较好地恢复图像微弱细节。     

Photoacoustic Tomography of Biological Sample Using Phase-controlled Focus Algorithm

Photoacoustic (PA) imaging is a new imaging modality, which converts pressure signals received by a scanning detector to a local distribution of electromagnetic absorption density. In this paper an experiment result of a photoacoustic tomography to depths of ～7 mm for a real tissue is presented, using a 532-nm pulse YAG laser. The time-resolved stress detection technique was used for PA signal detection with a high temporal resolution. A phase-controlled focus algorithm was used for image reconstruction. Images of different depth profiles in tissue were obtained. The depth resolution was 30 μm and could be up to 10 μm using a wide-band tranducer.     

Photoacoustic Tomography of Biological Sample Using Phase-controlled Focus Algorithm

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Is 'virtual histology' the next step after the 'virtual autopsy'? Magnetic resonance microscopy in forensic medicine

AIM: The study aimed to validate magnetic resonance microscopy (MRM) studies of forensic tissue specimens (skin samples with electric injury patterns) against the results from routine histology. METHODS AND RESULTS: Computed tomography and magnetic resonance imaging are fast becoming important tools in clinical and forensic pathology. This study is the first forensic application of MRM to the analysis of electric injury patterns in human skin. Three-dimensional high-resolution MRM images of fixed skin specimens provided a complete 3D view of the damaged tissues at the site of an electric injury as well as in neighboring tissues, consistent with histologic findings. The image intensity of the dermal layer in T2-weighted MRM images was reduced in the central zone due to carbonization or coagulation necrosis and increased in the intermediate zone because of dermal edema. A subjacent blood vessel with an intravascular occlusion supports the hypothesis that current traveled through the vascular system before arcing to ground. CONCLUSION: High-resolution imaging offers a noninvasive alternative to conventional histology in forensic wound analysis and can be used to perform 3D virtual histology.     

Real-time four-dimensional optical-resolution photoacoustic microscopy with Au nanoparticle-assisted subdiffraction-limit resolution

Photoacoustic microscopy (PAM) offers label-free, optical absorption contrast. A high-speed, high-resolution PAM system in an inverted microscope configuration with a laser pulse repetition rate of 100,000 Hz and a stationary ultrasonic transducer was built. Four-dimensional in vivo imaging of microcirculation in mouse skin was achieved at 18 three-dimensional volumes per second with repeated two-dimensional (2D) raster scans of 100 by 50 points. The corresponding 2D B-scan (50 A-lines) frame rate was 1800 Hz, and the one-dimensional A-scan rate was 90,000 Hz. The lateral resolution is 0.23 ± 0.03 μm for Au nanowire imaging, which is 2.0 times below the diffraction limit.     

Improving signal levels in intravital multiphoton microscopy using an objective correction collar

Multiphoton microscopy has enabled biologists to collect high-resolution images hundreds of microns into biological tissues, including tissues of living animals. While the depth of imaging exceeds that possible from any other form of light microscopy, multiphoton microscopy is nonetheless generally limited to depths of less than a millimeter. Many of the advantages of multiphoton microscopy for deep tissue imaging accrue from the unique nature of multiphoton fluorescence excitation. However, the quadratic relationship between illumination level and fluorescence excitation makes multiphoton microscopy especially susceptible to factors that degrade the illumination focus. Here we examine the effect of spherical aberration on multiphoton microscopy in fixed kidney tissues and in the kidneys of living animals. We find that spherical aberration, as evaluated from axial asymmetry in the point-spread function, can be corrected by adjustment of the correction collar of a water immersion objective lens. Introducing a compensatory positive spherical aberration into the imaging system decreases the depth-dependence of signal levels in images collected from living animals, increasing signal by up to 50%.     

Interference microscopy for three-dimensional imaging with wavelength-to-depth encoding

A novel interference microscope for three-dimensional (3D) imaging based on a wavelength-to-depth encoding technique is presented. Wavelength-to-depth encoding is realized by use of a diffractive lens and wavelength tuning. A high depth resolution of 0.71 mum is obtained with 0.90-N.A. objective lenses. Experimental measurements of a four-level grating are presented, and the results are found to be comparable with those obtained with a Dektak profilometer and a similar interference microscope that uses mechanical depth scanning. The system is promising for fast, noncontact, high-resolution 3D imaging.     

Dual-modal photoacoustic/CT imaging system

Photoacoustic(PA) tomography(PAT) breaks the barrier for high-resolution optical imaging in a strong lightscattering medium, having a great potential for both clinical implementation and small animal studies. However,many organs and tissues lack enough PA contrast or even hinder the propagation of PA waves. Therefore, it is challenging to interpret pure PAT images, especially three-dimensional(3 D) PA images for deep tissues, without enough structural information. To overcome this limitation, in this study, we integrated PAT with X-ray computed tomography(CT) in a standalone system. PAT provides optical contrast and CT gives anatomical information. We performed agar, tissue phantom, and animal studies, and the results demonstrated that PAT/CT imaging systems can provide accurate spatial registration of important complementary contrasts.     

Lensless high-resolution photoacoustic imaging scanner for in vivo skin imaging

We previously launched a high-resolution photoacoustic (PA) imaging scanner based on a unique lensless design for in vivo skin imaging. The design, imaging algorithm and characteristics of the system are described in this paper. Neither an optical lens nor an acoustic lens is used in the system. In the imaging head, four sensor elements are arranged quadrilaterally, and by checking the phase differences for PA waves detected with these four sensors, a set of PA signals only originating from a chromophore located on the sensor center axis is extracted for constructing an image. A phantom study using a carbon fiber showed a depth-independent horizontal resolution of 84.0 ± 3.5 µm, and the scan direction-dependent variation of PA signals was about ± 20%. We then performed imaging of vasculature phantoms: patterns of red ink lines with widths of 100 or 200 μm formed in an acrylic block co-polymer. The patterns were visualized with high contrast, showing the capability for imaging arterioles and venues in the skin. Vasculatures in rat burn models and healthy human skin were also clearly visualized in vivo.     

Microimaging of hairless rat skin by magnetic resonance at 900 MHz

Purpose

Quantitative imaging of the rat skin was performed using magnetic resonance imaging (MRI) at 900 MHz.

Materials and methods

A number of imaging techniques utilized for multiple contrast included magnetization transfer contrast, spin-lattice relaxation constant (T1-weighting), combination of T2-weighting with magnetic field inhomogeneity (T2*-weighting), magnetization transfer weighting and diffusion tensor weighting. These were used to obtain 2D slices and 3D multislice-multiecho images with high magnetic resonance contrast. These 2D and 3D imaging techniques were combined to achieve high-resolution MRI.

Results

Oil–water phantom showed distinct fat-water contrast. The dermis and epidermis, including the stratum corneum remnants, of nude rat skin were distinct due to their proton magnetic resonance as a result of proton interactions with the skin interstitial tissue. Combined details obtained from high-resolution, high-quality ex vivo skin images with different multicontrast characteristics generated better differentiation of skin layers, sublayers and significant correlation (r²=0.4927 for MRI area, r²=0.3068 for histology area; P<.0148) of MR data with co-registered histological areas of the epidermis as well as the hair follicle.

Conclusion

The multiple contrast approach provided a noninvasive ex vivo MRI visualization with semi-quantitative assessment of the major skin structures including the stratum corneum remnants, epidermis, hair, papillary dermis, reticular dermis and hypodermis.     

Extended depth of focus in tomographic phase microscopy using a propagation algorithm

Tomographic phase microscopy is a laser interferometry technique in which a 3D refractive index map of a biological sample is constructed from quantitative phase images collected at a set of illumination angles. Although the resulting tomographic images provide valuable information, their resolution declines at axial distances beyond about 1 microm from the focal plane. We describe an improved 3D reconstruction algorithm in which the field at the focal plane is numerically propagated to depths throughout the sample. Diffraction is thus incorporated, extending the depth of focus to more than 10 mum. Tomograms with improved focal depth are demonstrated for single HT29 cells.     

Tilted objects EFI extracted at once by 3D output of the angular spectrum method

Since many optical imaging systems suffer from a limited depth of focus, three-dimensional (3D) objects, or even flat objects tilted with respect to the optical axis, are imaged with only a portion of them in focus. Typically, to overcome this problem, in-focus areas are extracted from different images taken at different depths. The in-focus regions are thus merged together to build the extended focus image (EFI). In this work, we propose a quasi-automatic method for the EFI construction of tilted objects, extracted at once by 3D output of the Angular Spectrum Method (ASM) from a single digitally recorded hologram. Results show that our method can be effectively used for the correction of the anamorphism due to the reconstruction on a tilted plane with respect to the hologram one.     

The characteristics of three-dimensional skin imaging system by full-colored optical coherence tomography

In the present cosmetic market, the skin image obtained from a hand-held camera is two-dimensional (2-D). Due to insufficient penetration, only the skin surface can be detected, and thus phenomena in the dermis cannot be observed. To take the place of the conventional 2D camera, a new hand-held imaging system is proposed for three-dimensional (3-D) skin imaging. Featuring non-invasiveness, optical coherence tomography (OCT) has become one of the popular medical imaging techniques. The dermal images shown in OCT-related reports were mainly single-colored because of the use of a monotonic light source. With three original-colored beams applied in OCT, a full-colored image can be derived for dermatology. The penetration depth of the system ranges from 0.43 to 0.78 mm, sufficient for imaging of main tissues in the dermis. Colorful and non-invasive perspectives of deep dermal structure help to advance skin science, dermatology and cosmetology.     

High-resolution optical coherence microscopy for high-speed, in vivo cellular imaging

Optical coherence microscopy (OCM) is demonstrated with a high-speed, broadband, reflective-grating phase modulator and a femtosecond Ti:Al2O3 laser. The novel system design permits high-resolution OCM imaging in a new operating regime in which a short coherence gate is used to relax the requirement for high-numerical-aperture confocal axial sectioning. In vivo cellular imaging is demonstrated in the Xenopus laevis tadpole and in human skin with a 3-microm coherence gate and a 30-microm confocal gate. The ability to achieve cellular imaging with a lower numerical aperture should facilitate the development of miniaturized probes for in vivo imaging applications.     

Near-infrared laser scanning confocal microscopy and its application in bioimaging

Near-infrared (NIR) fluorescence imaging is an important imaging technology in deep-tissue biomedical imaging and related researches, due to the low absorption and scattering of NIR excitation and/or emission in biological tissues. Laser scanning confocal microscopy (LSCM) plays a significant role in the family of fluorescence microscopy. Due to the introduction of pinhole, it can provide images with optical sectioning, high signal-to-noise ratio and better spatial resolution. In this study, in order to combine the advantages of these two techniques, we set up a fluorescence microscopic imaging system, which can be named as NIR-LSCM. The system was based on a commercially available confocal microscope, utilizing a NIR laser for excitation and a NIR sensitive detector for signal collection. In addition, NIR fluorescent nanoparticles (NPs) were prepared, and utilized for fluorescence imaging of the ear and brain of living mice based on the NIR-LSCM system. The structure of blood vessels at certain depth could be visualized clearly, because of the high-resolution and large-depth imaging capability of NIR-LSCM.     

Holoscopy--holographic optical coherence tomography

Scanning optical coherence tomography (OCT) is limited in sensitivity and resolution by the restricted focal depth of the confocal detection scheme. Holoscopy, a combination of holography and Fourier-domain full-field OCT, is proposed as a way to detect photons from all depths of a sample volume simultaneously with uniform sensitivity and lateral resolution, even at high NAs. By using the scalar diffraction theory, as frequently applied in digital holographic imaging, we fully reconstruct the object field with depth-invariant imaging quality. In vivo imaging of human skin is demonstrated with an image quality comparable to conventionally scanned OCT.     

Method of bimodal photoacoustic and ultrasound microscopy for simultaneous structural and functional diagnostics of biotissues

The excitation of external electrode of focused photoacoustic (PA) detector by the backscattered laser radiation can ensure a cost-effective upgrade from single-modality PA microscopy to dual-modality PA/US imaging. In this work we approbated thermoelastic generation of probing ultrasonic pulses for bimodal photoacoustic (PA) and optically mediated Ultrasound (US) microscopy in vivo, using single optical pulse delivered from tunable laser to form both PA and US A-scans. The presented results of bimodal in vivo visualization of rat brain provide with the experimental evidence, that the proposed approach can be used for both functional and structural bioimaging simultaneously.     

Spectroscopic imaging of the brain with phased-array coils at 3.0 T

The goal of this study was to develop and evaluate high-resolution magnetic resonance spectroscopic imaging (MRSI) utilizing the gains in signal-to-noise ratio (SNR) provided by combining higher magnetic field with high-sensitivity phased-array (PA) coils. We investigated the maximum improvement in spatial resolution as small as 0.09 cm(3) for brain MRSI while maintaining adequate SNR and acquisition time. The use of low peak power, dual-band spectral-spatial pulses was also investigated for application to 3 T MRSI of the brain using the body coil for radiofrequency excitation and PA coils for signal reception.