Direct en-face, speckle-reduced images using angular-compounded MS-OCT

In this paper, an angular compounding method to achieve speckle contrast reduction in optical coherence tomography (OCT) imaging is explored in detail. The angular compounding approach involves collecting multiple images at different angles of incidence, registering the images to correct for induced distortions, and then incoherently summing the images to reduce speckle. The method was experimentally demonstrated with a spectrometer-based Master-Slave enhanced OCT instrument capable of directly generating en-face images. We have investigated the impact of the angular range and number of averaged frames on the degree of speckle artefact reduction, as well as the effect on image resolution and sharpness. The minimum angular step necessary to secure a sufficiently small speckle pattern correlation between the images has also been determined, and the method has subsequently been validated on a biological sample (potato cells).

All-fibre supercontinuum laser for in vivo multispectral PAM of lipids in the extended NIR

In this paper we demonstrate a compact all-fibre, high pulse energy supercontinuum laser that covers a spectral range from 1440 to 1870 nm with a 7 ns pulse duration and total energy of 18.3 µJ at a repetition rate of 100 kHz. Using the developed high-pulse energy source, we perform multi-spectral photoacoustic microscopy imaging of lipids, both ex vivo on adipose tissue and in vivo to study the development of Xenopus laevis tadpoles, using six different excitation bands over the first overtone transition of C-H vibration bonds (1650-1850 nm).

En-face optical coherence tomography/fluorescence endomicroscopy for minimally invasive imaging using a robotic scanner

We report a compact rigid instrument capable of delivering en-face optical coherence tomography (OCT) images alongside (epi)-fluorescence endomicroscopy (FEM) images by means of a robotic scanning device. Two working imaging channels are included: one for a one-dimensional scanning, forward-viewing OCT probe and another for a fiber bundle used for the FEM system. The robotic scanning system provides the second axis of scanning for the OCT channel while allowing the field of view (FoV) of the FEM channel to be increased by mosaicking. The OCT channel has resolutions of 25  /  60  μm (axial/lateral) and can provide en-face images with an FoV of 1.6  ×  2.7  mm2. The FEM channel has a lateral resolution of better than 8  μm and can generate an FoV of 0.53  ×  3.25  mm2 through mosaicking. The reproducibility of the scanning was determined using phantoms to be better than the lateral resolution of the OCT channel. Combined OCT and FEM imaging were validated with ex-vivo ovine and porcine tissues, with the instrument mounted on an arm to ensure constant contact of the probe with the tissue. The OCT imaging system alone was validated for in-vivo human dermal imaging with the handheld instrument. In both cases, the instrument was capable of resolving fine features such as the sweat glands in human dermal tissue and the alveoli in porcine lung tissue.

Quantitative assessment of rat bone regeneration using complex master-slave optical coherence tomography

The aim of this work is to evaluate the capability of an in-house developed multimodal complex master slave (CMS) enhanced swept source (SS) optical coherence tomography (OCT) imaging instrument to analyze the increase in the quantity and the improvement of the quality of newly-formed bone using low level laser therapy (LLLT). Bone formation is quantitatively assessed in 5 mm cylindrical defects made in the calvaria part of the skull of living rats. Samples are divided in three study groups: A, a negative control group, for which the natural healing process of the defect is investigated; B, a positive control group, for which bovine graft is used to stimulate bone formation, and C, a study group, in which bovine graft is added to the created defects and LLLT is applied throughout the entire healing period. The animals are sacrificed after 14, 21, and 30 days, and the samples are imaged using the multimodal CMS/SS-OCT instrument.

Optical Coherence Tomography Investigations and Modeling of the Sintering of Ceramic Crowns

Dental prostheses are sintered in ovens that sometimes suffer from a loss of calibration. This can lead to variations of the sintering temperature outside the range recommended by the manufacturer. Stress and even fractures in dental ceramics may occur, and this leads to the necessity to rebuild the dental construct. The aim of this work is to monitor the quality of sintering processes using an established biomedical imaging technique—optical coherence tomography (OCT). Conventional current procedures imply the fabrication of supplemental samples that add to the expenses and are only evaluated visually. To our knowledge, we were the first to propose the use of OCT, a non-destructive method that brings objectivity for such assessments, focusing, in a previous study, on metal ceramic dental prostheses. Here, a different material, pressed ceramics, is considered, while we propose a quantitative assessment of the results—using reflectivity profiles of en-face (i.e., constant-depth) OCT images of sintered samples. The results for both the pressed ceramics and metal ceramics prostheses are discussed by obtaining the analytic functions of their reflectivity profiles. A multi-parametric analysis demonstrates the best parameter to characterize the loss of calibration of dental ovens. Rules-of-thumb are extracted; producing dental prostheses with efects can thus be avoided.

Fast spectrally encoded Mueller optical scanning microscopy

Mueller microscopes enable imaging of the optical anisotropic properties of biological or non-biological samples, in phase and amplitude, at sub-micrometre scale. However, the development of Mueller microscopes poses an instrumental challenge: the production of polarimetric parameters must be sufficiently quick to ensure fast imaging, so that the evolution of these parameters can be visualised in real-time, allowing the operator to adjust the microscope while constantly monitoring them. In this report, a full Mueller scanning microscope based on spectral encoding of polarization is presented. The spectrum, collected every 10 μs for each position of the optical beam on the specimen, incorporates all the information needed to produce the full Mueller matrix, which allows simultaneous display of all the polarimetric parameters, at the unequalled rate of 1.5 Hz (for an image of 256 × 256 pixels). The design of the optical blocks allows for the real-time display of linear birefringent images which serve as guidance for the operator. In addition, the instrument has the capability to easily switch its functionality from a Mueller to a Second Harmonic Generation (SHG) microscope, providing a pixel-to-pixel matching of the images produced by the two modalities. The device performance is illustrated by imaging various unstained biological specimens.

Down-conversion en-face optical coherence tomography

We present an optical coherence tomography (OCT) method that can deliver an en-face OCT image from a sample in real-time, irrespective of the tuning speed of the swept source. The method, based on the master slave interferometry technique, implements a coherence gate principle by requiring that the optical path difference (OPD) between the arms of an imaging interferometer is the same with the OPD in an interrogating interferometer. In this way, a real-time en-face OCT image can originate from a depth in the sample placed in the imaging interferometer, selected by actuating on the OPD in the interrogating interferometer, while laterally scanning the incident beam over the sample. The generation of the en-face image resembles time domain OCT, with the difference that here the signal is processed based on spectral domain OCT. The optoelectronic processor operates down-conversion of the chirped radio frequency signal delivered by the photo-detector. The down-conversion factor is equal to the ratio of the maximum frequency of the photo-detected signal due to an OPD value matching the coherence length of the swept source, to the sweeping rate. This factor can exceed 106 for long coherence swept sources.

Complex master-slave for long axial range swept-source optical coherence tomography

Using complex master-slave interferometry, we demonstrate extended axial range optical coherence tomography for two commercially available swept sources, well beyond the limit imposed by their k-clocks. This is achieved without k-domain re-sampling and without engaging any additional Mach-Zehnder interferometer providing a k-clock signal to the digitizer. An axial imaging range exceeding 17 mm with an attenuation of less than 30 dB is reported using two commercially available swept sources operating at 1050 nm and a 100 kHz repetition rate. This procedure has more than trebled the range achievable using the k-clock signal provided by the manufacturers. An analysis is presented on the impact that the digitization has on the axial range and resolution of the system.

Recovering distance information in spectral domain interferometry

This work evaluates the performance of the Complex Master Slave (CMS) method, that processes the spectra at the interferometer output of a spectral domain interferometry device without involving Fourier transforms (FT) after data acquisition. Reliability and performance of CMS are compared side by side with the conventional method based on FT, phase calibration with dispersion compensation (PCDC). We demonstrate that both methods provide similar results in terms of resolution and sensitivity drop-off. The mathematical operations required to produce CMS results are highly parallelizable, allowing real-time, simultaneous delivery of data from several points of different optical path differences in the interferometer, not possible via PCDC.

Group refractive index and group velocity dispersion measurement by complex master slave interferometry

This paper demonstrates that the complex master slave interferometry (CMSI) method used in spectral domain interferometry (SDI) can efficiently be used for accurate refractive index and group velocity dispersion measurements of optically transparent samples. For the first time, we demonstrate the relevance of the phase information delivered by CMSI for dispersion evaluations with no need to linearize data. The technique proposed here has been used to accurately measure the group refractive index and the group velocity dispersion of a strong dispersive sample (SF6 glass), and a weak dispersive one (distilled water). The robustness of the technique is demonstrated through the manipulation of several sets of experimental data.