We present a novel technique that uses hybrid digital holography to observe three-dimensional morphological changes in cardiac organoids and measure heart rates simultaneously. By reconstructing the real-time spatial topology, we capture dynamic morphological changes in three-dimensional cardiac organoids, especially in the central region where traditional optical imaging methods are limited. We analyze the correlation between the phase analysis results of digital holography and the corresponding electronic signal amplitude with heartbeat data. Fluorescent modes can also be used to investigate heart rate, ROS activation, and organic cell activation. In addition, dark field mode allows you to simultaneously evaluate the distribution of nanoparticles about 100 nm in size on the heart surface and their effect on the heart rate. Our work demonstrates the potential of hybrid digital holography as an innovative optical analysis method in heart and tissue analysis, providing valuable insights into the relationship between cardiac tissue dynamics and heart rate
Histopathology is the fundamental microscopic examination tool in many domains including cancer diagnosis and surgical oncology, and various fields. Recently, the UV-PAM is powerful imaging system for label-free histology-like imaging. In histopathology, the nuclei in cancer cells have typical morphological features, such as irregular shapes and large sizes. Alternatively, UV-PAM can obtain unstrained cell nuclei with specific, positive, and high-contrast images. UV-PAM is useful histological images of unstained cell nuclei with specific, positive, and high-image contrast. By using the 266 nm light, high contrast photoacoustic images can be obtained. In the recent UV-PAM, the hardware has been faced a fundamental limitation to laser source especially. There is no available commercial nanosecond (ns) laser that has a high repetition rate operation of hundreds of kHz at 266 nm. Here, we develop a stable high-repetition rate 266 nm source using the basic second harmonic generation (SHG) phenomenon. This stable 266 nm source is based on a simplified architecture using single-pass SHG with BBO crystal.
Retinal images technology such as optical coherence tomography (OCT), OCT-angiography (OCTA) and fluorescein angiography (FA) are an important for diagnosing eye diseases. As retinal imaging technology continues to evolve, standard test methods and optical phantoms for demonstrating image quality and performance also need to be developed and advanced. We used mixture solution of polydimethylsiloxzne (PDMS) and different titanium dioxide (TiO2) to develop a more realistic retinal phantom. The retinal phantom consists of two multilayer membranes, a base plate with retinal curvature, and three microfluidic channels. In addition, we developed retinal phantom for fluorescence imaging using micro-fluorescence beads. We could obtain OCT, OCTA, and fluorescence images.
Observing micro-vessels in conjunctiva could be used not only for diagnosing conjunctival diseases including conjunctivitis and pterygium but also as biomarkers for circulatory diseases. Many research teams have developed compact imaging and auto-analysis systems to simplify the conventional slit lamp as well as enhance the analysis process. The imaging system, previously developed by our research team, corrects eye motion in imaging windows through image registration and template matching. The developed system quantifies blood flow velocity using the sequence of motion-corrected images. This study compares estimated flow velocity and the fluid's actual velocity using the experimental phantom comprising transparent hoses and fluid including beads corresponding to red blood cells. The flow velocities are calculated using the Hagen-Poiseuille equation, and the flow rates generated by the syringe pump. The pump applies three kinds of flow rates to generate flow velocity variations and the estimated velocities are linearly proportional to these variations. In addition, the phantom has a random motion to mimic the fixational eye movements within the range of the healthy subject's angular eye motion. Through these experiments, we verified the previously developed flow velocity measurement system having percent errors under 3% by comparing estimated flow velocities with actual flow velocities. The system's accuracy, especially under conditions without artificial motion, is over 98.5%. These experiments can provide the supporting background for feasibility and accuracy in a further clinical study in conjunctival microcirculation.
3D cell models that are induced to interact and grow in a 3D environment are formed up to several hundred micrometers. Since cells grow in all directions, an imaging system that can show molecular images using fluorescence as well as morphological structures without damaging the structure of the cell model is needed. We have developed a multimodal optical microscopy imaging system that combines extended depth of focus-optical coherence microscopy (EDOF-OCM) and two-photon fluorescence microscopy (TPFM) for more accurate measurement of 3D cells. This imaging system was able to acquire merged images without moving the spheroids, and the efficiency of the imaging systems was compared using optical coherence tomography (OCT), OCM, and confocal fluorescence microscopy(CFM).
KEYWORDS: 3D modeling, Optical coherence tomography, Imaging systems, Tumors, 3D image processing, Bessel beams, 3D metrology, 3D acquisition, Visualization, Tissues
3D cell models indicate stronger similarities in vivo than monolayer cell culture, and thus is raising awareness as an important tool for evaluating biological phenomena, drug action, and mechanism of diseases. This paper compared two imaging systems for accurate measurements of the morphological structure and volume quantification of 3D tumor spheroids. The two imaging systems are OCT using a scan lens and EDOF-OCM, which uses a Bessel beam to expand the depth of field. Human hepatoma cells were used for spheroid formation. To compare the effectiveness of the imaging system, we measured the spheroid structure according to cell number per spheroid and growth per day. Although both systems were able to acquire the morphological structure and quantify volume of 3D tumor spheroids the OCM using the Bessel beam was able to more accurately measure the overall structure of the spheroid without surface reflection.
The retinal imaging system, such as optical coherence tomography (OCT), OCT-angiography (OCTA) and fluorescein angiography (FA), is the important subject for ophthalmic. The use of such equipment continues to increase and retinal phantoms have also been developed to evaluate and modify its performance and image quality. In this presentation, we will show methods and results of a retinal phantom that can evaluate the optical performance of OCT, OCTA and FA. We implemented superficial vascular networks and full retinal layers which has curvature. We could obtain cross-sectional OCT images and en-face OCTA images using lab-made OCT system. In addition, FA image could also be obtained through sodium fluorescein dye injection.
Optical coherence tomography (OCT) is one of imaging technologies to diagnose ophthalmic diseases and to monitor progresses of treatments. Therefore, OCT and OCT-angiography (OCTA) have actively developed in the field of ophthalmology, and retinal phantoms for evaluation of performance of OCT and OCTA also have been studied. In this presentation, we will show methods and results of a retinal phantom that can evaluate the optical performance of OCT and OCTA. The retinal phantom not only mimicked superficial vascular networks more realistically, but also implemented retinal layers with curvature. It consists of two microfluidic channels corresponding to superficial and deep retinal vessels, two multi-layered thin films, and base plate with retinal curvature. Each was attached with oxygen plasma for 1-min and fluorinated ethylene propylene (FEP) tubes was connected to the inlet and outlet holes. We could obtain cross-sectional OCT images and en-face OCTA images using lab-made OCT system.
We developed an eye phantom that mimic retinal layers and blood vessels to evaluation of retinal OCT-angiography and fundus angiography. The phantom was made of polydimethylsiloxane and titanium dioxide using spin coater and microelectromechanical system (MEMS) technology.
We present a dual-wavelength laser using stimulated Raman scattering (SRS) effect which have high pulse repetition rate for fast functional photoacoustic (PA) imaging. This laser has a high pulse repetition rate of 300 kHz and high pulse energy more than 200 nJ. We periodically modulated the electro-optic modulator voltage from 0 to 168 V to switch the polarization of the output light. Two different pulse lights separated by polarization switching were used to generate different SRS peaks using Raman fibers with lengths of 5 and 20 m. The operating wavelength of this laser was switched to 545 nm and 603 nm using SRS effect and polarization switching. Wavelength dependent fast functional PA images of blood vessels and gold nanorods were obtained using a dual-wavelength switchable SRS pulsed-laser.
Optical coherence tomography (OCT) is one of useful diagnostic devices for retinal diseases. Recently, optical coherence tomography based angiography (OCTA) extends the OCT applications from structural images to functional images by enabling blood vessel networks mapping. As the use of OCT and OCTA increases in ophthalmology, it is necessary to develop retinal phantoms for performance evaluation of clinical OCT devices for retinal imaging.
In this study, we have developed a retinal layer-mimicking phantom including microfluidic channels to assess OCT and OCTA image quality and to evaluate software accuracy. The phantom is constructed of thin scattering films based on polydimethylsiloxane (PDMS) and titanium dioxide (TiO2) powder. We adjusted TiO2 particle concentration in PDMS for matching intensity of retinal OCT images. Particles were well dispersed throughout PDMS using a probe tip sonicator. Mixed PDMSs were carried out spin coating on a glass slide to make thin films. Before spin coating, silane was applied to the glass substrate to provide a hydrophobic coating for easily removing of cured PDMS. After spinning, thin filmed PDMSs were cured. Microfluidic channels were also made with PDMSs mixed with TiO2 powder, and were designed with sizes of 50 um, 100 um, and 200 um. thin films of cured PDMS were stacked on microfluidic channels. We used diluted dye liquids containing microbeads to occur optically scattered liquid flow like blood vessel. Dye liquids containing microbeads were flowed into an inlet port of the channel through a syringe pump
Finally, we successfully obtained cross-sectional volumetric OCT and OCTA images of completed retinal phantom using lab-made OCT system and clinical OCT system.
Optical-resolution photoacoustic microscopy (OR-PAM), has been widely used and studied as noninvasive and in-vivo imaging technique, can achieve a high resolution and high contrast image. OR-PAM is combined with optical absorption contrast and detection of acoustic wave generated by thermal expansion. Recently, nanoparticles and dyes have been used as contrast agents of OR-PAM. To obtain functional OR-PAM image such as a distribution image of blood vessels and nanoparticles, a tunable dye laser or optical parametric oscillator (OPO) should be needed at more two wavelength. However, because these lasers have a low pulse repetition rate (10 Hz ~ 10 kHz), a functional OR-PAM image with real-time display has been limited.
In our previous study, we demonstrated high-speed OR-PAM using an Ytterbium fiber laser and a graphics processing unit (GPU) technique at 300 kHz-pulse repetition rates. Although this Ytterbium fiber laser has a high pulse repetition rate, it is not comfortable for functional imaging owing to lasing at only single wavelength. Therefore, in this study, we used a high-speed interlaced illumination method at 532 nm and 1064 nm for real-time display functional OR-PAM. For high-speed interlaced illumination of two wavelength, we applied second harmonic generation effect and a high-speed optical switching using an electro-optic modulator. Therefore, we could obtain maximum amplitude projection (MAP) images about distributions of blood vessels and nanoparticles, simultaneously, with 500 x 500 pixels and a real-time display of approximately 0.5 fps.
Optical coherence tomography (OCT) is a nondestructive imaging technique offering high temporal and spatial resolution, which makes it a natural choice for assessing tissue mechanical properties. We have developed methods to mechanically analyze the compliance of the rabbit trachea in vivo using tissue deformations induced by tidal breathing, offering a unique tool to assess the behavior of the airways during their normal function. Four-hundred images were acquired during tidal breathing with a custom-built endoscopic OCT system. The surface of the tissue was extracted from a set of these images via image processing algorithms, filtered with a bandpass filter set at respiration frequency to remove cardiac and probe motion, and compared to ventilatory pressure to calculate wall compliance. These algorithms were tested on elastic phantoms to establish reliability and reproducibility. The mean tracheal wall compliance (in five animals) was 1.3±0.3×10−5 (mm Pa)−1. Unlike previous work evaluating airway mechanics, this new method is applicable in vivo, noncontact, and loads the trachea in a physiological manner. The technique may have applications in assessing airway mechanics in diseases such as asthma that are characterized by significant airway remodeling.
We demonstrated high-speed spectral domain polarization-sensitive optical coherence tomography (SD-PSOCT)
using a single InGaAs line-scan camera and an optical switch at 1.3-μm region. The polarization-sensitive
low coherence interferometer in the system was based on the original free-space PS-OCT system published by
Hee et al. The horizontal and vertical polarization light rays split by polarization beam splitter were delivered
and detected via an optical switch to a single spectrometer by turns instead of dual spectrometers. The SD-PSOCT
system had an axial resolution of 8.2 μm, a sensitivity of 101.5 dB, and an acquisition speed of 23,496 Alines/s. We obtained the intensity, phase retardation, and fast axis orientation images of a biological tissue. In
addition, we calculated the averaged axial profiles of the phase retardation in human skin.
We propose high-speed spectral domain polarization-sensitive optical coherence tomography (SD-PS-OCT) using a single camera and a 1×2 optical switch at the 1.3-µm region. The PS-low coherence interferometer used in the system is constructed using free-space optics. The reflected horizontal and vertical polarization light rays are delivered via an optical switch to a single spectrometer by turns. Therefore, our system costs less to build than those that use dual spectrometers, and the processes of timing and triggering are simpler from the viewpoints of both hardware and software. Our SD-PS-OCT has a sensitivity of 101.5 dB, an axial resolution of 8.2 µm, and an acquisition speed of 23,496 A-scans per second. We obtain the intensity, phase retardation, and fast axis orientation images of a rat tail tendon ex vivo.
We built a high-speed, real-time spectral domain optical coherence tomography (SD-OCT) system at the 1.3 μm region
using an InGaAs line-scan camera with 1024 pixels and 46.99 klines/s. In SD-OCT, the actual spatial resolution can be
different from the theoretical one due to the large bandwidth of the light source and the finite number of detector pixels,
especially for a long center wavelength. We calculated and compared the axial resolutions obtained from the point spread
function and the physical pixel size of the OCT image. We found that the axial resolution of the SD-OCT system could
be limited in the 1.3 μm region if the depth range becomes large.
We demonstrate that changes in the degree of polarization (DOP) depend on changes in the scattering coefficient, and they can be quantified by using a polarization-sensitive optical coherence tomography (PS-OCT) system. We test our hypothesis using liquid and solid phantoms made from Intralipid suspensions and gelatin, respectively. We also quantify the DOP changes with depth caused by changes in the concentration of scatterers in the liquid and solid phantoms. It is clearly shown that the DOP change has a linear relationship with the scattering change. In our previous study, we showed that the axial slope of the DOP is different between normal and pathologic cervical tissues. Our results demonstrate that the quantification of the axial DOP slope can be used for the systematic diagnosis of certain tissue pathology.
We use polarization-sensitive optical coherence tomography (PS-OCT) to monitor the wound healing process in vitro and in vivo, which are affected by various drugs. Five rabbit subjects are used for in vitro studies and another five are used for in vivo studies. The in vitro studies are conducted to compare the PS-OCT images with histopathology. For each subject, three biopsy lesions are created on each ear: one site is not treated (control); the second site is treated with sphingosylphosphorylcholine, which is expected to promote healing; and the last is administered with tetraacetylphytosphingosine, which negatively affects the healing process. Each site is examined with a PS-OCT system at 1, 4, 7, 10, and 14- days after wound generation. The variations of phase retardation values caused by the collagen morphology changes on wound sites are quantified for all cases. Our results suggest that PS-OCT may be a useful tool for visualization of collagen fiber regeneration and for quantification of various drug effects during the wound healing process.
We use polarization sensitive optical coherence tomography (PS-OCT) to monitor wound healing processes in-vitro and in-vivo, which are affected by various drugs. Five rabbit subjects are used for the in-vitro studies and another five are used for in-vivo studies. The in-vitro studies are conducted to compare the PS-OCT images with histopathology. For each subject, three biopsy lesions are created on each ear: one site is not treated (control), the second site is treated with sphingosyl phosphoryl choline (SPC), which is known to promote healing, and the last is administered with tetra acetyl phytosphingo sine (TAPS), which negatively affects the healing process. Each site is examined with a PS-OCT system and conventional histopathology at 1-, 4-, 7-, 10-, and 14-days after wound generation. The phase retardation values are quantified for all cases and our results suggest that PS-OCT may be a useful tool for visualization of collagen fiber regeneration during the healing process; therefore, various drug effects can be noninvasively monitored.
We measure the skin wrinkle topology by means of low coherence interferometry (LCI), which forms the basis of the optical coherence tomography (OCT). The skin topology obtained using LCI and corresponding 2-D fast Fourier transform allow quantification of skin wrinkles. It took approximately 2 minutes to obtain 2.1 mm x 2.1 mm topological image with 4 um and 16 um resolutions in axial and transverse directions, respectively. Measurement examples show the particular case of skin contour change after-wrinkle cosmeceutical treatments and atopic dermatitis
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