Multimodality endoscopic optical coherence tomography and fluorescence imaging technology for visualization of layered architecture and subsurface microvasculature.

Endoscopic imaging technologies, such as endoscopic optical coherence tomography (OCT) and near-infrared fluorescence, have been used to investigate vascular and morphological changes as hallmarks of early cancer in the gastrointestinal tract. Here we developed a high-speed multimodality endoscopic OCT and fluorescence imaging system. Using this system, the architectural morphology and vasculature of the rectum wall were obtained simultaneously from a Sprague Dawley rat in vivo. This multimodality imaging strategy in a single imaging system permits the use of a single imaging probe, thereby improving prognosis by early detection and reducing costs.

Coaxial excitation longitudinal shear wave measurement for quantitative elasticity assessment using phase-resolved optical coherence elastography.

Optical coherence elastography (OCE) is an emerging imaging modality for the assessment of mechanical properties in soft tissues. Transverse shear wave measurements using OCE can quantify the elastic moduli perpendicular to the force direction, however, missing the elastic information along the force direction. In this study, we developed coaxial excitation longitudinal shear wave measurements for quantification of elastic moduli along the force direction using M-scans. Incorporating Rayleigh wave measurements using non-coaxial lateral scans into longitudinal shear wave measurements, directionally dependent elastic properties can be quantified along the force direction and perpendicular to the force direction. Therefore, the reported system has the capability to image elasticity of anisotropic biological tissues.

Linear-in-wavenumber swept laser with an acousto-optic deflector for optical coherence tomography.

We report a novel linear-in-wavenumber (k-linear) swept laser source based on an acousto-optic deflector (AOD). The AOD-based optical filter includes an acousto-optic deflector and a reflection grating. The laser may tune k linearly in wavenumber over time due to its appropriate configuration and is favorable for fast imaging because it avoids data resampling and recalibration, as are required in conventional swept source optical coherence tomography (SS-OCT). We achieved k-linearity with Pearson's r correlation coefficients of 0.99995 without and 0.99997 with optimization. The laser has a tuning range of 50 nm, a 3 dB swept range of 42 nm (FWHM), output power of 2.56 mW, 6 dB sensitivity roll-off depth of 0.941 mm, and central wavelength of 1064 nm at a scanning rate of ∼20 kHz. Scanning rate as high as ∼400 kHz is also achieved for this laser with the tuning range 49 nm, swept linearity of 0.99990, output power of 2.30 mW, and a 6 dB sensitivity roll-off depth 0.550 mm. SS-OCT imaging with linear-in-wavenumber swept laser is also demonstrated.

Spectral-domain optical coherence tomography with a Fresnel spectrometer.

We propose a novel spectral-domain optical coherence tomography (SD-OCT) equipped with a Fresnel spectrometer, which utilizes a Fresnel zone plate (FZP) as both dispersion and focusing optics and thus spreads the spectral interferogram evenly in wavenumber domain because of the proportional relation between the focal length of the FZP and the wavenumber. With no need of the conversion calculation from wavelength to wavenumber in conventional SD-OCT, this new design is favorable for fast imaging with high resolution. As only a FZP and CCD are used, the Fresnel spectrometer is simple and compact. It is experimentally shown that its performance is as good as that of numerical interpolation in conventional SD-OCT. Imaging of bio-tissue by Fresnel SD-OCT is also demonstrated.

Compressed sensing with linear-in-wavenumber sampling in spectral-domain optical coherence tomography.

We propose a novel method called compressed sensing with linear-in-wavenumber sampling (k-linear CS) to retrieve an image for spectral-domain optical coherence tomography (SD-OCT). An array of points that is evenly spaced in wavenumber domain is sampled from an original interferogram by a preset k-linear mask. Then the compressed sensing based on l1 norm minimization is applied on these points to reconstruct an A-scan data. To get an OCT image, this method uses less than 20% of the total data as required in the typical process and gets rid of the spectral calibration with numerical interpolation in traditional CS-OCT. Therefore k-linear CS is favorable for high speed imaging. It is demonstrated that the k-linear CS has the same axial resolution performance with ~30 dB higher signal-to-noise ratio (SNR) as compared with the numerical interpolation. Imaging of bio-tissue by SD-OCT with k-linear CS is also demonstrated.

Integrated pulse scope for tunable generation and intrinsic characterization of structured femtosecond laser.

Numerous techniques have been demonstrated for effective generation of orbital angular momentum-carrying radiation, but intracavity generation of continuously tunable pulses in the femtosecond regime remains challenging. Even if such a creation was realized, the generated pulses-like all pulses in reality-are complex and transitory objects that can only be comprehensively characterized via multidimensional spaces. An integrated lasing system that generates pulses while simultaneously quantifies them can achieve adaptive pulse tailoring. Here, we report a femtosecond pulse scope that unifies vector vortex mode-locked lasing and vectorial quantification. With intracavity-controlled Pancharatnam-Berry phase modulation, continuous and ergodic generation of spirally polarized states along a broadband higher-order Poincaré sphere was realized. By intrinsically coupling a two-dimensional polarization-sensitive time-scanning interferometer to the laser, multidimensional spatiotemporal features of the pulse were further visualized. The proposed methodology paves the way for design optimization of ultrafast optics by integrating complex femtosecond pulse generation and structural customization, facilitating its applications in optical physics research and laser-based manufacturing.

PMN-PT/Epoxy 1-3 composite based ultrasonic transducer for dual-modality photoacoustic and ultrasound endoscopy.

Endoscopic dual-modality photoacoustic (PA) and ultrasound (US) imaging has the capability of providing morphology and molecular information simultaneously. An ultrasonic transducer was applied for detecting PA signals and performing US imaging which determines the sensitivity and performance of a dual-modality PA/US system. In our study, a miniature single element 32-MHz lead magnesium niobate-lead titanate (PMN-PT) epoxy 1-3 composite based ultrasonic transducer was developed. A miniature endoscopic probe based on this transducer has been fabricated. Using the dual modality PA/US system with a PMN-PT/epoxy 1-3 composite based ultrasonic transducer, phantom and in vivo animal studies have been conducted to evaluate the performance. The preliminary results show enhanced bandwidths of the new ultrasonic transducer and improved signal-to-noise ratio of PA and US images of rat colorectal wall compared with PMN-PT and lead zirconate titanate (PZT) composite based ultrasonic transducers.

Deep imaging in highly scattering media by combining reflection matrix measurement with Bessel-like beam based optical coherence tomography.

Multiple scattering in biomedical tissue limits the imaging depth within a range of 1-2 mm for conventional optical imaging techniques. To extend the imaging depth into the scattering medium, a computational method based on the reflection matrix measurement has been developed to retrieve the singly back-scattered signal light from the dominant detrimental multiple-scattered background. After applying singular value decomposition on the measured matrix in the post-process, the target image underneath the turbid media is clearly recovered. To increase the depth of focus of the incident light by elongating the focal spot along the optical axis, a digital grating pattern is specially designed and displayed on a phase-only spatial light modulator to generate the Bessel-like beam for lateral point scanning. According to the results, the depth of focus is increased up to 2.4 mm which is much longer than the value of ∼50 µm obtained by using the conventional focused Gaussian beam, leading to a deeper penetration depth due to the self-healing feature of the Bessel-like beam. In addition, generation of the Bessel-like beam simplifies the axial scanning process by getting rid of the need to mechanically translate the focal zone along the optical axis of an objective with a high numerical aperture. By combining this method with an optical coherence tomography system with a low coherence light source, a depth-resolved optical image is obtained underneath a highly turbid medium.

Quantitative angle-insensitive flow measurement using relative standard deviation OCT.

Incorporating different data processing methods, optical coherence tomography (OCT) has the ability for high-resolution angiography and quantitative flow velocity measurements. However, OCT angiography cannot provide quantitative information of flow velocities, and the velocity measurement based on Doppler OCT requires the determination of Doppler angles, which is a challenge in a complex vascular network. In this study, we report on a relative standard deviation OCT (RSD-OCT) method which provides both vascular network mapping and quantitative information for flow velocities within a wide range of Doppler angles. The RSD values are angle-insensitive within a wide range of angles, and a nearly linear relationship was found between the RSD values and the flow velocities. The RSD-OCT measurement in a rat cortex shows that it can quantify the blood flow velocities as well as map the vascular network in vivo.

Optical computing for optical coherence tomography.

We propose an all-optical Fourier transformation system for real-time massive data processing in high speed optical coherence tomography (OCT). In the so-called optical computing OCT, fast Fourier transformation (FFT) of A-scan signal is optically processed in real time before being detected by photoelectric detector. Therefore, the processing time for interpolation and FFT in traditional Fourier domain OCT can be dramatically eliminated. A processing rate of 10 mega-A-scans/second was experimentally achieved, which is, to our knowledge, the highest speed for OCT imaging. Due to its fiber based all-optical configuration, this optical computing OCT system is ideal for ultrahigh speed volumetric OCT imaging in clinical application.

Ultrahigh-speed optical coherence tomography utilizing all-optical 40 MHz swept-source.

We present an ultrahigh-speed optical coherence tomography (OCT) based on an all-optical swept-source with an A-scan rate of 40 MHz. The inertia-free swept-source, which has its output power of 41.2 mW and tuning range of 40 nm and high scan linearity in wavenumber with Pearson's correlation coefficients r of 0.9996, consists of a supercontinuum laser, an optical band-pass filter, a linearly chirped fiber Bragg grating, an erbium-doped fiber amplifier, and two buffer stages. With sensitivity of 87 dB, high-speed OCT imaging of biological tissue in vivo is also demonstrated.

Tiny endoscopic optical coherence tomography probe driven by a miniaturized hollow ultrasonic motor.

We present an endoscopic probe for optical coherence tomography (OCT) equipped with a miniaturized hollow ultrasonic motor that rotates the objective lens and provides an internal channel for the fiber to pass through, enabling 360 deg unobstructed circumferential scanning. This probe has an outer diameter of 1.5 mm, which is ultra-small for motorized probes with an unobstructed view in distal scanning endoscopic OCT. Instead of a mirror or prism, a customized aspheric right-angle lens is utilized, leading to an enlargement of the numerical aperture and thus high transverse resolution. Spectral-domain OCT imaging of bio-tissue and a phantom are demonstrated with resolution of 7.5 µm(axial)×6.6 µm(lateral) and sensitivity of 96 dB.

Understanding three-dimensional spatial relationship between the mouse second polar body and first cleavage plane with full-field optical coherence tomography.

The morphogenetic relationship between early patterning and polarity formation is of fundamental interest and remains a controversial issue in preimplantation embryonic development. We use a label-free three-dimensional (3-D) imaging technique of full-field optical coherence tomography (FF-OCT) successfully for the first time to study the dynamics of developmental processes in mouse preimplantation lives. Label-free 3-D subcellular time-lapse images are demonstrated to investigate 3-D spatial relationship between the second polar body (2PB) and the first cleavage plane. By using FF-OCT together with quantitative study, we show that only 25% of the predicted first cleavage planes, defined by the apposing plane of two pronuclei, pass through the 2PB. Also only 27% of the real cleavage planes pass through the 2PB. These results suggest that the 2PB is not a convincing spatial cue for the event of the first cleavage. Our studies demonstrate the feasibility of FF-OCT in providing new insights and potential breakthroughs to the controversial issues of early patterning and polarity in mammalian developmental biology.

Noninvasive three-dimensional live imaging methodology for the spindles at meiosis and mitosis.

The spindle plays a crucial role in normal chromosome alignment and segregation during meiosis and mitosis. Studying spindles in living cells noninvasively is of great value in assisted reproduction technology (ART). Here, we present a novel spindle imaging methodology, full-field optical coherence tomography (FF-OCT). Without any dye labeling and fixation, we demonstrate the first successful application of FF-OCT to noninvasive three-dimensional (3-D) live imaging of the meiotic spindles within the mouse living oocytes at metaphase II as well as the mitotic spindles in the living zygotes at metaphase and telophase. By post-processing of the 3-D dataset obtained with FF-OCT, the important morphological and spatial parameters of the spindles, such as short and long axes, spatial localization, and the angle of meiotic spindle deviation from the first polar body in the oocyte were precisely measured with the spatial resolution of 0.7 µm. Our results reveal the potential of FF-OCT as an imaging tool capable of noninvasive 3-D live morphological analysis for spindles, which might be useful to ART related procedures and many other spindle related studies.

Label-free subcellular 3D live imaging of preimplantation mouse embryos with full-field optical coherence tomography.

Early patterning and polarity is of fundamental interest in preimplantation embryonic development. Label-free subcellular 3D live imaging is very helpful to its related studies. We have developed a novel system of full-field optical coherence tomography (FF-OCT) for noninvasive 3D subcellular live imaging of preimplantation mouse embryos with no need of dye labeling. 3D digitized embryos can be obtained by image processing. Label-free 3D live imaging is demonstrated for the mouse embryos at various typical preimplantation stages with a spatial resolution of 0.7 [micro sign]m and imaging rate of 24 fps. Factors that relate to early patterning and polarity, such as pronuclei in zygote, shapes of zona pellucida, location of second polar body, cleavage planes, and the blastocyst axis, can be quantitatively measured. The angle between the two second cleavage planes is accurately measured to be 87 deg. It is shown that FF-OCT provides a potential breakthrough for early patterning, polarity formation, and many other preimplantation-related studies in mammalian developmental biology.