共查询到20条相似文献,搜索用时 62 毫秒
1.
Bumju Kim Myeongsu Na Soohyun Park Kitae Kim Jung-Hoon Park Euiheon Chung Sunghoe Chang Ki Hean Kim 《Biomedical optics express》2021,12(4):2328
Open-top light-sheet microscopy (OT-LSM) is a specialized microscopic technique for high throughput cellular imaging of large tissue specimens including optically cleared tissues by having the entire optical setup below the sample stage. Current OT-LSM systems had relatively low axial resolutions by using weakly focused light sheets to cover the imaging field of view (FOV). In this report, open-top axially swept LSM (OTAS-LSM) was developed for high-throughput cellular imaging with improved axial resolution. OTAS-LSM swept a tightly focused excitation light sheet across the imaging FOV using an electro tunable lens (ETL) and collected emission light at the focus of the light sheet with a camera in the rolling shutter mode. OTAS-LSM was developed by using air objective lenses and a liquid prism and it had on-axis optical aberration associated with the mismatch of refractive indices between air and immersion medium. The effects of optical aberration were analyzed by both simulation and experiment, and the image resolutions were under 1.6µm in all directions. The newly developed OTAS-LSM was applied to the imaging of optically cleared mouse brain and small intestine, and it demonstrated the single-cell resolution imaging of neuronal networks. OTAS-LSM might be useful for the high-throughput cellular examination of optically cleared large tissues. 相似文献
2.
High resolution three-dimensional photoacoutic tomography with CCD-camera based ultrasound detection
A photoacoustic tomograph based on optical ultrasound detection is demonstrated, which is capable of high resolution real-time projection imaging and fast three-dimensional (3D) imaging. Snapshots of the pressure field outside the imaged object are taken at defined delay times after photoacoustic excitation by use of a charge coupled device (CCD) camera in combination with an optical phase contrast method. From the obtained wave patterns photoacoustic projection images are reconstructed using a back propagation Fourier domain reconstruction algorithm. Applying the inverse Radon transform to a set of projections recorded over a half rotation of the sample provides 3D photoacoustic tomography images in less than one minute with a resolution below 100 µm. The sensitivity of the device was experimentally determined to be 5.1 kPa over a projection length of 1 mm. In vivo images of the vasculature of a mouse demonstrate the potential of the developed method for biomedical applications.OCIS codes: (110.0110) Imaging systems, (170.5120) Photoacoustic imaging, (170.3880) Medical and biological imaging, (170.6960) Tomography 相似文献
3.
Transient intrinsic optical responses associated with neural activation offer an attractive strategy for dynamic imaging of neural activity, and may provide a noninvasive methodology for imaging of retinal function. Here we demonstrate the feasibility of near infrared imaging of fast intrinsic optical changes in isolated frog retina activated by visible light. Using a photodiode detector in a transmitted light geometry, we routinely measured dynamic transmitted optical responses in single passes, at the level of one part in 10(4) of background light. Rapid CCD image sequences acquired with transmitted light (bright field) illumination disclosed larger fractional responses and showed evidence of multiple response components with both negative- and positive-going signals with different timecourses. Dark field imaging further enhanced the contrast and sensitivity of optical measures of neural activation. High-resolution imaging disclosed optical responses in single pixels often exceeding 5%, of background light, allowing dynamic imaging at the resolution of single cells, in single passes. Fast optical signals are closely related to identified response components of the electroretinogram. Optical responses showed complex but consistent spatial organization from frame to frame. Our experimental results and theoretical analysis suggest that the optical responses may result from dynamic volume changes corresponding to ion and water flow across the cell membrane, directly associated with the electrophysiological response. 相似文献
4.
Lingling Chen Yuriy Alexandrov Sunil Kumar Natalie Andrews Margaret J. Dallman Paul M. W. French James McGinty 《Biomedical optics express》2015,6(4):1253-1261
We describe an angular multiplexed imaging technique for 3-D in vivo cell tracking of sparse cell distributions and optical projection tomography (OPT) with superior time-lapse resolution and a significantly reduced light dose compared to volumetric time-lapse techniques. We demonstrate that using dual axis OPT, where two images are acquired simultaneously at different projection angles, can enable localization and tracking of features in 3-D with a time resolution equal to the camera frame rate. This is achieved with a 200x reduction in light dose compared to an equivalent volumetric time-lapse single camera OPT acquisition with 200 projection angles. We demonstrate the application of this technique to mapping the 3-D neutrophil migration pattern observed over ~25.5 minutes in a live 2 day post-fertilisation transgenic LysC:GFP zebrafish embryo following a tail wound.OCIS codes: (170.2520) Fluorescence microscopy, (170.6900) Three-dimensional microscopy, (170.6920) Time-resolved imaging, (170.3010) Image reconstruction techniques 相似文献
5.
Michael Münter Mario Pieper Tabea Kohlfaerber Ernst Bodenstorfer Martin Ahrens Christian Winter Robert Huber Peter Knig Gereon Hüttmann Hinnerk Schulz-Hildebrandt 《Biomedical optics express》2021,12(10):6024
Volumetric imaging of dynamic processes with microscopic resolution holds a huge potential in biomedical research and clinical diagnosis. Using supercontinuum light sources and high numerical aperture (NA) objectives, optical coherence tomography (OCT) achieves microscopic resolution and is well suited for imaging cellular and subcellular structures of biological tissues. Currently, the imaging speed of microscopic OCT (mOCT) is limited by the line-scan rate of the spectrometer camera and ranges from 30 to 250 kHz. This is not fast enough for volumetric imaging of dynamic processes in vivo and limits endoscopic application. Using a novel CMOS camera, we demonstrate fast 3-dimensional OCT imaging with 600,000 A-scans/s at 1.8 µm axial and 1.1 µm lateral resolution. The improved speed is used for imaging of ciliary motion and particle transport in ex vivo mouse trachea. Furthermore, we demonstrate dynamic contrast OCT by evaluating the recorded volumes rather than en face planes or B-scans. High-speed volumetric mOCT will enable the correction of global tissue motion and is a prerequisite for applying dynamic contrast mOCT in vivo. With further increase in imaging speed and integration in flexible endoscopes, volumetric mOCT may be used to complement or partly replace biopsies. 相似文献
6.
Omkar D. Supekar Andrew Sias Sean R. Hansen Gabriel Martinez Graham C. Peet Xiaoyu Peng Victor M. Bright Ethan G. Hughes Diego Restrepo Douglas P. Shepherd Cristin G. Welle Juliet T. Gopinath Emily A. Gibson 《Biomedical optics express》2022,13(4):2530
We present a high-resolution miniature, light-weight fluorescence microscope with electrowetting lens and onboard CMOS for high resolution volumetric imaging and structured illumination for rejection of out-of-focus and scattered light. The miniature microscope (SIMscope3D) delivers structured light using a coherent fiber bundle to obtain optical sectioning with an axial resolution of 18 µm. Volumetric imaging of eGFP labeled cells in fixed mouse brain tissue at depths up to 260 µm is demonstrated. The functionality of SIMscope3D to provide background free 3D imaging is shown by recording time series of microglia dynamics in awake mice at depths up to 120 µm in the brain. 相似文献
7.
Jürgen Dammers Markus Axer David Gräßel Christoph Palm Karl Zilles Katrin Amunts Uwe Pietrzyk 《NeuroImage》2010,49(2):1241-1248
Polarized light imaging (PLI) enables the evaluation of fiber orientations in histological sections of human postmortem brains, with ultra-high spatial resolution. PLI is based on the birefringent properties of the myelin sheath of nerve fibers. As a result, the polarization state of light propagating through a rotating polarimeter is changed in such a way that the detected signal at each measurement unit of a charged-coupled device (CCD) camera describes a sinusoidal signal. Vectors of the fiber orientation defined by inclination and direction angles can then directly be derived from the optical signals employing PLI analysis. However, noise, light scatter and filter inhomogeneities interfere with the original sinusoidal PLI signals. We here introduce a novel method using independent component analysis (ICA) to decompose the PLI images into statistically independent component maps. After decomposition, gray and white matter structures can clearly be distinguished from noise and other artifacts. The signal enhancement after artifact rejection is quantitatively evaluated in 134 histological whole brain sections. Thus, the primary sinusoidal signals from polarized light imaging can be effectively restored after noise and artifact rejection utilizing ICA. Our method therefore contributes to the analysis of nerve fiber orientation in the human brain within a micrometer scale. 相似文献
8.
Richard J. Colchester Edward Z. Zhang Paul C. Beard Adrien E. Desjardins 《Biomedical optics express》2022,13(7):4047
All-optical ultrasound (OpUS), where ultrasound is both generated and received using light, has emerged as a modality well-suited to highly miniaturised applications. In this work we present a proof-of-concept OpUS transducer built onto a single optical fibre with a highly miniaturised lateral dimension (<0.8 mm). A key innovation was to use a dual-clad optical fibre (DCF) to provide multimode light for ultrasound generation and single mode light for ultrasound reception. The transducer comprised a proximal section of DCF spliced to a short section of single mode fibre (SMF). Multimode light was outcoupled at the splice joint and guided within a square capillary to provide excitation for ultrasound generation. Whilst single mode light was guided to the distal tip of the SMF to a plano-concave microresonator for ultrasound reception. The device was capable of generating ultrasound with pressures >0.4 MPa and a corresponding bandwidth >27 MHz. Concurrent ultrasound generation and reception from the transducer enabled imaging via motorised pull-back allowing image acquisition times of 4 s for an aperture of 20 mm. Image resolution was as low as ~50 µm and 190 µm in the axial and lateral extents, respectively, without the need for image reconstruction. Porcine aorta was imaged ex vivo demonstrating detailed ultrasound images. The unprecedented level of miniaturisation along with the high image quality produced by this device represents a radical new paradigm for minimally invasive imaging. 相似文献
9.
Endocytoscopy is an optical biopsy technique which uses a miniaturized camera to capture white light microscopy images through an endoscope. We have developed an alternative design that instead relays images to an external camera via a coherent fiber bundle. In this paper we characterize the device and demonstrate microscopy of porcine tissue ex vivo. One advantage of our approach is the ease with which other bundle-compatible imaging modalities can be deployed simultaneously. We show this by acquiring quasi-simultaneous endocytoscopy and fluorescence confocal endomicroscopy images through a single fiber bundle. This opens up possibilities for multi-modal endomicroscopy, combining white light and fluorescence imaging.OCIS codes: (170.2150) Endoscopic imaging, (110.0180) Microscopy 相似文献
10.
Photoacoustic (PA) imaging is an emerging hybrid imaging modality that combines rich optical spectroscopic contrast and high ultrasonic resolution, and thus holds tremendous promise for a wide range of pre-clinical and clinical applications. Compact and affordable light sources such as light-emitting diodes (LEDs) and laser diodes (LDs) are promising alternatives to bulky and expensive solid-state laser systems that are commonly used as PA light sources. These could accelerate the clinical translation of PA technology. However, PA signals generated with these light sources are readily degraded by noise due to the low optical fluence, leading to decreased signal-to-noise ratio (SNR) in PA images. In this work, a spatiotemporal singular value decomposition (SVD) based PA denoising method was investigated for these light sources that usually have low fluence and high repetition rates. The proposed method leverages both spatial and temporal correlations between radiofrequency (RF) data frames. Validation was performed on simulations and in vivo PA data acquired from human fingers (2D) and forearm (3D) using a LED-based system. Spatiotemporal SVD greatly enhanced the PA signals of blood vessels corrupted by noise while preserving a high temporal resolution to slow motions, improving the SNR of in vivo PA images by 90.3%, 56.0%, and 187.4% compared to single frame-based wavelet denoising, averaging across 200 frames, and single frame without denoising, respectively. With a fast processing time of SVD (∼50 µs per frame), the proposed method is well suited to PA imaging systems with low-energy excitation light sources for real-time in vivo applications. 相似文献
11.
This paper proposes a novel approach for high-resolution light field microscopy imaging by using a camera array. In this approach, we apply a two-stage relay system for expanding the aperture plane of the microscope into the size of an imaging lens array, and utilize a sensor array for acquiring different sub-apertures images formed by corresponding imaging lenses. By combining the rectified and synchronized images from 5 × 5 viewpoints with our prototype system, we successfully recovered color light field videos for various fast-moving microscopic specimens with a spatial resolution of 0.79 megapixels at 30 frames per second, corresponding to an unprecedented data throughput of 562.5 MB/s for light field microscopy. We also demonstrated the use of the reported platform for different applications, including post-capture refocusing, phase reconstruction, 3D imaging, and optical metrology.OCIS codes: (180.6900) Three-dimensional microscopy, (110.1758) Computational imaging, (170.0110) Imaging systems, (100.3010) Image reconstruction techniques 相似文献
12.
Derek Nankivil Gar Waterman Francesco LaRocca Brenton Keller Anthony N. Kuo Joseph A. Izatt 《Biomedical optics express》2015,6(11):4516-4528
We describe the first handheld, swept source optical coherence tomography (SSOCT) system capable of imaging both the anterior and posterior segments of the eye in rapid succession. A single 2D microelectromechanical systems (MEMS) scanner was utilized for both imaging modes, and the optical paths for each imaging mode were optimized for their respective application using a combination of commercial and custom optics. The system has a working distance of 26.1 mm and a measured axial resolution of 8 μm (in air). In posterior segment mode, the design has a lateral resolution of 9 μm, 7.4 mm imaging depth range (in air), 4.9 mm 6dB fall-off range (in air), and peak sensitivity of 103 dB over a 22° field of view (FOV). In anterior segment mode, the design has a lateral resolution of 24 μm, imaging depth range of 7.4 mm (in air), 6dB fall-off range of 4.5 mm (in air), depth-of-focus of 3.6 mm, and a peak sensitivity of 99 dB over a 17.5 mm FOV. In addition, the probe includes a wide-field iris imaging system to simplify alignment. A fold mirror assembly actuated by a bi-stable rotary solenoid was used to switch between anterior and posterior segment imaging modes, and a miniature motorized translation stage was used to adjust the objective lens position to correct for patient refraction between −12.6 and + 9.9 D. The entire probe weighs less than 630 g with a form factor of 20.3 x 9.5 x 8.8 cm. Healthy volunteers were imaged to illustrate imaging performance.OCIS codes: (110.4500) Optical coherence tomography, (170.4460) Ophthalmic optics and devices, (080.3620) Lens system design, (170.0110) Imaging systems, (170.5755) Retina scanning, (170.4470) Ophthalmology 相似文献
13.
Imaging the retinal vasculature offers a surrogate view of systemic vascular health, allowing
noninvasive and longitudinal assessment of vascular pathology. The earliest anomalies in vascular
disease arise in the microvasculature, however current imaging methods lack the spatiotemporal
resolution to track blood flow at the capillary level. We report here on novel imaging technology
that allows direct, noninvasive optical imaging of erythrocyte flow in human retinal capillaries.
This was made possible using adaptive optics for high spatial resolution (1.5 μm), sCMOS
camera technology for high temporal resolution (460 fps), and tunable wavebands from a broadband
laser for maximal erythrocyte contrast. Particle image velocimetry on our data sequences was used to
quantify flow. We observed marked spatiotemporal variability in velocity, which ranged from 0.3 to
3.3 mm/s, and changed by up to a factor of 4 in a given capillary during the 130 ms imaging period.
Both mean and standard deviation across the imaged capillary network varied markedly with time, yet
their ratio remained a relatively constant parameter (0.50 ± 0.056). Our observations concur
with previous work using less direct methods, validating this as an investigative tool for the study
of microvascular disease in humans.OCIS codes: (110.1080) Active or adaptive optics, (120.7250) Velocimetry, (170.1470) Blood or tissue constituent monitoring, (170.2655) Functional monitoring and imaging, (170.4470) Ophthalmology 相似文献
14.
Rowan W. Sanderson Qi Fang Andrea Curatolo Aiden Taba Helen M. DeJong Fiona M. Wood Brendan F. Kennedy 《Biomedical optics express》2021,12(6):3117
Smartphones are now integral to many telehealth services that provide remote patients with an improved diagnostic standard of care. The ongoing management of burn wounds and scars is one area in which telehealth has been adopted, using video and photography to assess the repair process over time. However, a current limitation is the inability to evaluate scar stiffness objectively and repeatedly: an essential measurement for classifying the degree of inflammation and fibrosis. Optical elastography detects mechanical contrast on a micrometer- to millimeter-scale, however, typically requires expensive optics and bulky imaging systems, making it prohibitive for wide-spread adoption in telehealth. More recently, a new variant of optical elastography, camera-based optical palpation, has demonstrated the capability to perform elastography at low cost using a standard digital camera. In this paper, we propose smartphone-based optical palpation, adapting camera-based optical palpation by utilizing a commercially available smartphone camera to provide sub-millimeter resolution imaging of mechanical contrast in scar tissue in a form factor that is amenable to telehealth. We first validate this technique on a silicone phantom containing a 5 × 5 × 1 mm3 embedded inclusion, demonstrating comparative image quality between mounted and handheld implementations. We then demonstrate preliminary in vivo smartphone-based optical palpation by imaging a region of healthy skin and two scars on a burns patient, showing clear mechanical contrast between regions of scar tissue and healthy tissue. This study represents the first implementation of elastography on a smartphone device, extending the potential application of elastography to telehealth. 相似文献
15.
Jiheun Ryu Ungyo Kang Joon Woo Song Junyoung Kim Jin Won Kim Hongki Yoo Bomi Gweon 《Biomedical optics express》2021,12(9):5452
Optical microscopy has been widely used in biomedical research as it provides photophysical and photochemical information of the target in subcellular spatial resolution without requiring physical contact with the specimen. To obtain a deeper understanding of biological phenomena, several efforts have been expended to combine such optical imaging modalities into a single microscope system. However, the use of multiple light sources and detectors through separated beam paths renders previous systems extremely complicated or slow for in vivo imaging. Herein, we propose a novel high-speed multimodal optical microscope system that simultaneously visualizes five different microscopic contrasts, i.e., two-photon excitation, second-harmonic generation, backscattered light, near-infrared fluorescence, and fluorescence lifetime, using a single femtosecond pulsed laser. Our proposed system can visualize five modal images with a frame rate of 3.7 fps in real-time, thereby providing complementary optical information that enhances both structural and functional contrasts. This highly photon-efficient multimodal microscope system enables various properties of biological tissues to be assessed. 相似文献
16.
We demonstrate feasibility of endoscopic imaging of Cerenkov light originated when charged nuclear particles, emitted from radionuclides, travel through a biological tissue of living subjects at superluminal velocity. The endoscopy imaging system consists of conventional optical fiber bundle/ clinical endoscopes, an optical imaging lens system, and a sensitive low-noise charge coupled device (CCD) camera. Our systematic studies using phantom samples show that Cerenkov light from as low as 1 μCi of radioactivity emitted from (18)F-Fluorodeoxyglucose (FDG) can be coupled and transmitted through conventional optical fibers and endoscopes. In vivo imaging experiments with tumor bearing mice, intravenously administered with (18)F-FDG, further demonstrated that Cerenkov luminescence endoscopy is a promising new tool in the field of endoscopic molecular imaging. 相似文献
17.
Phillip Bedggood Alexis Ceecee Britten-Jones Lauren N. Ayton Andrew Metha 《Biomedical optics express》2022,13(10):5311
The optical density of visual pigment can be measured by imaging the dark-adapted eye while bleaching with visible light. This measurement can be made for individual photoreceptor cells using adaptive optics; however, activation of the phototransduction cascade imparts rapid changes in phase that modulate the signal via optical interference. This limits utility because data must be averaged over many experimental runs. Here we used a “flood” illuminated adaptive optics system at 4000 fps, bright light to achieve rapid bleaching, and broad illumination bandwidth to mitigate interference effects. Data were super-resolved using the natural motion of the eye to overcome the reduced pixel resolution of the ultrafast camera. This approach was applied to classify the trichromatic cone photoreceptor mosaic at a single fixation locus within the foveal region of 3 healthy subjects. Subjects were dark adapted for 6 minutes to replenish cone photopigment. This was followed either directly by imaging at 555 ± 50 nm, or by first pre-adapting the retina to 700 nm light to preferentially deplete “L” cone pigment. A total of 3,252 cones were classified as either “S”, “M”, or “L” type based on clustering of the intensity data observed under these two conditions. Mean classification probability ranged from 99.3 to 99.8%, with individual cell probabilities exceeding 95% in 97.0 to 99.2% of cones. Accuracy of cone classification peaked when using the first 10-30 ms of data, with significant reductions in accuracy noted with the inclusion of data from later times. Our results show that rapid bleaching and data acquisition significantly improve the robustness of cell-resolved densitometry. 相似文献
18.
Monolithic Vernier tuned super-structure grating distributed Bragg reflector (SSG-DBR) lasers are expected to become one of the most promising sources for swept source optical coherence tomography (SS-OCT) with a long coherence length, reduced sensitivity roll-off, and potential capability for a very fast A-scan rate. However, previous implementations of the lasers suffer from four main problems: 1) frequencies deviate from the targeted values when scanned, 2) large amounts of noise appear associated with abrupt changes in injection currents, 3) optically aliased noise appears due to a long coherence length, and 4) the narrow wavelength coverage of a single chip limits resolution. We have developed a method of dynamical frequency tuning, a method of selective data sampling to eliminate current switching noise, an interferometer to reduce aliased noise, and an excess-noise-free connection of two serially scanned lasers to enhance resolution to solve these problems. An optical frequency comb SS-OCT system was achieved with a sensitivity of 124 dB and a dynamic range of 55-72 dB that depended on the depth at an A-scan rate of 3.1 kHz with a resolution of 15 μm by discretely scanning two SSG-DBR lasers, i.e., L-band (1.560-1.599 μm) and UL-band (1.598-1.640 μm). A few OCT images with excellent image penetration depth were obtained.OCIS codes: (170.4500) Optical coherence tomography, (170.3880) Medical and biological imaging, (170.4470) Ophthalmology 相似文献
19.
Hassan Dibaji Md Nasful Huda Prince Yating Yi Hu Zhao Tonmoy Chakraborty 《Biomedical optics express》2022,13(9):4990
Axially swept light sheet microscopy (ASLM) is an emerging technique that enables isotropic, subcellular resolution imaging with high optical sectioning capability over a large field-of-view (FOV). Due to its versatility across a broad range of immersion media, it has been utilized to image specimens that may range from live cells to intact chemically cleared organs. However, because of its design, the performance of ASLM-based microscopes is impeded by a low detection signal and the maximum achievable frame-rate for full FOV imaging. Here we present a new optical concept that pushes the limits of ASLM further by scanning two staggered light sheets and simultaneously synchronizing the rolling shutter of a scientific camera. For a particular peak-illumination-intensity, this idea can make ASLMs image twice as fast without compromising the detection signal. Alternately, for a particular frame rate our method doubles the detection signal without requiring to double the peak-illumination-power, thereby offering a gentler illumination scheme compared to tradition single-focus ASLM. We demonstrate the performance of our instrument by imaging fluorescent beads and a PEGASOS cleared-tissue mouse brain. 相似文献
20.
We describe a high-speed camera system for frequency domain imaging suitable for applications such as in vivo diffuse optical imaging and fluorescence lifetime imaging. 14-bit images are acquired at 2 gigapixels per second and analyzed with real-time pipeline processing using field programmable gate arrays (FPGAs). Performance of the camera system has been tested both for RF-modulated laser imaging in combination with a gain-modulated image intensifier and a simpler system based upon an LED light source. System amplitude and phase noise are measured and compared against theoretical expressions in the shot noise limit presented for different frequency domain configurations. We show the camera itself is capable of shot noise limited performance for amplitude and phase in as little as 3 ms, and when used in combination with the intensifier the noise levels are nearly shot noise limited. The best phase noise in a single pixel is 0.04 degrees for a 1 s integration time. 相似文献