Ultrahigh-resolution full-field optical coherence tomography

We have developed a white-light interference microscope for ultrahigh-resolution full-field optical coherence tomography of biological media. The experimental setup is based on a Linnik-type interferometer illuminated by a tungsten halogen lamp. En face tomographic images are calculated by a combination of interferometric images recorded by a high-speed CCD camera. Spatial resolution of 1.8 microm x 0.9 microm (transverse x axial) is achieved owing to the extremely short coherence length of the source, the compensation of dispersion mismatch in the interferometer arms, and the use of relatively high-numerical-aperture microscope objectives. A shot-noise-limited detection sensitivity of 90 dB is obtained in an acquisition time per image of 4 s. Subcellular-level images of plant, animal, and human tissues are presented.     

Aperture effect correction in spectroscopic full-field optical coherence tomography

Spatially resolved spectroscopic optical coherence tomography (OCT) has been demonstrated to be a convenient tool for spectral analysis in turbid media. For a full-field OCT configuration using a Mirau objective in the visible range, we found that the effective numerical aperture varies over the field of view, leading to field-dependent spectral shifts in the reconstructed spectra. Interferograms recorded with quasi-monochromatic lights are theoretically fitted with a general Mirau interference formula, and we propose a numerical correction method for white-light spectroscopy. The method is then tested successfully for the measure of the reflectivity of a plane gold sample.     

Inverse scattering for frequency-scanned full-field optical coherence tomography

Full-field optical coherence tomography (OCT) is able to image an entire en face plane of scatterers simultaneously, but typically the focus is scanned through the volume to acquire three-dimensional structure. By solving the inverse scattering problem for full-field OCT, we show it is possible to computationally reconstruct a three-dimensional volume while the focus is fixed at one plane inside the sample. While a low-numerical-aperture (NA) OCT system can tolerate defocus because the depth of field is large, for high NA it is critical to correct for defocus. By deriving a solution to the inverse scattering problem for full-field OCT, we propose and simulate an algorithm that recovers object structure both inside and outside the depth of field, so that even for high NA the focus can be fixed at a particular plane within the sample without compromising resolution away from the focal plane.     

Sub-4-micron full-field optical coherence tomography on a budget

We evaluate the construction and performance of an ultra-low-cost full-field optical coherence tomography (FF-OCT) instrument, also known as an optical coherence microscope. Although the cost of construction of the instrument from off-the-shelf parts is at least ten times lower than those of commercial products, sub-4-micron axial and lateral resolutions are obtained, albeit at the cost of higher acquisition times. Standard test samples are imaged and the performance of the instrument is evaluated. The device is found to be useful in measuring length, dispersion and group refractive index as well. Suggestions for bettering performance are discussed.     

Signal-to-noise ratio study of full-field fourier-domain optical coherence tomography

We report a new approach in optical coherence tomography (OCT) called full-field Fourier-domain OCT (3F-OCT). A three-dimensional image of a sample is obtained by digital reconstruction of a three-dimensional data cube, acquired with a Fourier holography recording system, illuminated with a swept source. We present a theoretical and experimental study of the signal-to-noise ratio of the 3F-OCT approach versus serial image acquisition (flying-spot OCT) approach.     

High-resolution full-field optical coherence tomography with a Linnik microscope

We describe an original microscope for high-resolution optical coherence tomography applications. Our system is based on a Linnik interference microscope with high-numerical-aperture objectives. Lock-in detection of the interference signal is achieved in parallel on a CCD by use of a photoelastic birefringence modulator and full-field stroboscopic illumination with an infrared LED. Transverse cross-section (en-face, or XY) images can be obtained in real time with better than 1-microm axial (Z) resolution and 0.5-microm transverse (XY) resolution. A sensitivity of approximately 80 dB is reached at a 1-image/s acquisition rate, which allows tomography in scattering media such as biological tissues.     

Imaging of oocyte development using ultrahigh-resolution full-field optical coherence tomography

In this paper, a white-light full-field optical coherence tomography is developed to provide three-dimensional imaging of the development of a mouse embryo with ultrahigh-resolution. Spatial resolution of 1.8 μm×1.12 μm (transverse×axial) is achieved owing to the extremely short coherence length of the light source and optimized compensation of dispersion mismatch. A shot-noise limited detection sensitivity of 80 dB is obtained at an acquisition time of 5 seconds per image. To enable in vivo imaging of the mouse embryo development, a homemade incubator is applied to provide appropriate CO2 concentration, temperature, and humidity. An electronic light shutter is used to control the light source in order to avoid unnecessary exposure to the embryo development when the sample is not being scanned. To demonstrate our method, in vivo time series two-dimensional images of the in vitro fertilization process of mouse oocytes at the germinal vesicles stage are presented.     

Cartilage thickness measurements from optical coherence tomography

We describe a new semiautomatic image processing method for detecting the cartilage boundaries in optical coherence tomography (OCT). In particular, we focus on rabbit cartilage since this is an important animal model for testing both chondroprotective agents and cartilage repair techniques. The novel boundary-detection system presented here consists of (1) an adaptive filtering technique for image enhancement and speckle reduction, (2) edge detection, and (3) edge linking by graph searching. The procedure requires several steps and can be automated. The quantitative measurements of cartilage thickness on OCT images correlated well with measurements from histology.     

Image restoration method based on Hilbert transform for full-field optical coherence tomography

A full-field optical coherence tomography (FF-OCT) system utilizing a simple but novel image restoration method suitable for a high-speed system is demonstrated. An en-face image is retrieved from only two phase-shifted interference fringe images through using the mathematical Hilbert transform. With a thermal light source, a high-resolution FF-OCT system having axial and transverse resolutions of 1 and 2.2 microm, respectively, was implemented. The feasibility of the proposed scheme is confirmed by presenting the obtained en-face images of biological samples such as a piece of garlic and a gold beetle. The proposed method is robust to the error in the amount of the phase shift and does not leave residual fringes. The use of just two interference images and the strong immunity to phase errors provide great advantages in the imaging speed and the system design flexibility of a high-speed high-resolution FF-OCT system.     

Image enhancement for multilayer information retrieval by using full-field optical coherence tomography

When a full-field optical coherence tomography (OCT) system is used to extract tomographic images from a multilayer information carrier, the resulting images may suffer from interlayer modulations and parasitic patterns derived from interference fringes. We describe and analyze these negative influences that degrade the quality of extracted tomographic images and propose practical algorithms and methods to minimize them. The emphasis of the discussion will be the removal of the parasitic fringes produced by the imperfection of a CCD camera. The simulative and experimental results of image enhancement for multilayer tomography extraction using full-field OCT are provided.     

Speckle statistics in optical coherence tomography

The two previously reported calculations of the amplitude distribution of speckles in optical coherence tomography, each based on a different mathematical formulation, yield different results. We show that a modification of an initial assumption in one of the formulations leads to equivalent results.     

Interferometer for optical coherence tomography

We describe a new interferometer setup for optical coherence tomography (OCT). The interferometer is based on a fiber arrangement similar to Young's two-pinhole interference experiment with spatial coherent and temporal incoherent light. Depth gating is achieved detection of the interference signal on a linear CCD array. Therefore no reference optical delay scanning is needed. The interference signal, the modulation of the signal, the axial resolution, and the depth range are derived theoretically and compared with experiments. The dynamic range of the setup is compared with OCT sensors in the time domain. To our knowledge, the first images of porcine brain and heart tissue and human skin are presented.     

Parallel optical coherence tomography system

We present the design and procedures for implementing a parallel optical coherence tomography (POCT) imaging system that can be adapted to an endoscopic format. The POCT system consists of a single mode fiber (SMF) array with multiple reduced diameter (15 microm) SMFs in the sample arm with 15 microm center spacing between fibers. The size of the array determines the size of the transverse imaging field. Electronic scanning eliminates the need for mechanically scanning in the lateral direction. Experimental image data obtained with this system show the capability for parallel axial scan acquisition with lateral resolution comparable to mechanically scanned optical coherence tomography systems.     

Characterisation of scaffold architecture and tendons using optical coherence tomography and polarisation-sensitive optical coherence tomography

Scaffolds play an important role in the generation of functional tissues using tissue-engineering techniques. To generate highly organised tissue, scaffolds must have specific internal and external architectures. Here, optical coherence tomography (OCT) is exploited to characterise the architectures of various scaffolds, in particular scaffolds which have been fabricated to support the formation of uniaxially orientated collagen bundle for use in tendon tissue engineering. In parallel, a polarisation-sensitive OCT (PSOCT) has been built to assess the collagen fibre organisation in human tendon and monitor the growth of engineering tendon constructs online and non-destructively. The impact of mechanical stimuli on the modulation of tendon tissue formation and organisation was also assessed. It is shown that conventional OCT is capable of characterising scaffold architecture and the pore size, porosity or microchannel dimension can be determined quantitatively and qualitatively. PSOCT generated birefringence images of human tendon and demonstrated that low birefringence images, associated with fewer microstructural variations, correlated to the presence of scar tissue or degenerated tissue; whereas the tissue-engineered tendon exhibited lower degree of birefringence.     

High-sensitivity determination of birefringence in turbid media with enhanced polarization-sensitive optical coherence tomography

Polarization-sensitive optical coherence tomography provides high-resolution cross-sectional characterization of birefringence in turbid media. Weakly birefringent biological tissues such as the retinal nerve fiber layer (RNFL) require advanced speckle noise reduction for high-sensitivity measurement of form birefringence. We present a novel method for high-sensitivity birefringence quantification by using enhanced polarization-sensitive optical coherence tomography (EPS-OCT) and introduce the polarimetric signal-to-noise ratio, a mathematical tool for analyzing speckle noise in polarimetry. Multiple incident polarization states and non-linear fitting of normalized Stokes vectors allow determination of retardation with +/-1 degrees uncertainty with invariance to unknown unitary polarization transformations. Results from a weakly birefringent turbid film and in vivo primate RNFL are presented. In addition, we discuss the potential of EPS-OCT for noninvasive quantification of intracellular filamentous nanostructures, such as neurotubules in the RNFL that are lost during the progression of glaucoma.     

Temporal coherence and time-frequency distributions in spectroscopic optical coherence tomography

Traditional analysis of spectroscopic optical coherence tomography (SOCT) signals is limited by an uncertainty relationship between time (depth) and frequency (wavelength). The use of a bilinear time-frequency distribution for analysis, such as those that compose Cohen's class of functions, may provide a way to avoid this limitation. Here we present the relationship between traditional SOCT analysis and the relevant Cohen class functions: the Wigner and Choi-Williams distributions. While cross terms that arise in these bilinear time-frequency distributions have been viewed as an artifact, here we identify these terms with temporal coherence, which contains significant information about the signal through phase relationships. The utility of time-frequency distributions is illustrated through analysis of calculated signals.     

High-resolution frequency-domain second-harmonic optical coherence tomography

We used continuum generated in an 8.5 cm long fiber by a femtosecond Yb fiber laser to improve threefold the axial resolution of frequency domain second-harmonic optical coherence tomography (SH-OCT) to 12 microm. The acquisition time was shortened by more than 2 orders of magnitude compared to the time-domain SH-OCT. The system was applied to image biological tissue of fish scales, pig leg tendon, and rabbit eye sclera. Highly organized collagen fibrils can be visualized in the recorded images. Polarization dependence on the SH has been used to obtain polarization resolved images.     

Inverse scattering for optical coherence tomography

Inverse scattering theory for optical coherence tomography (OCT) is developed. The results are used to produce algorithms to resolve three-dimensional object structure, taking into account the finite beam width, diffraction, and defocusing effects. The resolution normally achieved only in the focal plane of the OCT system is shown to be available for all illuminated depths in the object without moving the focal plane. Spatially invariant resolution is verified with numerical simulations and indicates an improvement of the high-resolution cross-sectional imaging capabilities of OCT.     

线聚焦光学相干层析术的研究

提出了一种高速光学相干层析(OCT)成像技术方案。利用柱面镜的成像特性将传统OCT的点聚焦成像模式改变为线聚焦成像模式,从而降低二维OCT图像的扫描维数,达到提高成像速度的目的。利用ZEMAX光学软件对系统进行光线追迹获得光束经过柱面镜后的聚焦情况。随后采用635nm的激光光源和柱面镜构建了实验系统,实验结果很好地验证了光线追迹仿真结果。