Tethered capsule en face optical coherence tomography for imaging Barrett's oesophagus in unsedated patients.

OBJECTIVE: Barrett's oesophagus (BE) screening outside the endoscopy suite can identify patients for surveillance and reduce mortality. Tethered capsule optical coherence tomography (OCT) can volumetrically image oesophageal mucosa in unsedated patients and detect features of BE. We investigated ultrahigh-speed tethered capsule swept-source OCT (SS-OCT), improved device design, developed procedural techniques and measured capsule contact, longitudinal pullback non-uniformity and patient toleration. DESIGN: OCT was performed in 16 patients prior to endoscopic surveillance/treatment. Unsedated patients swallowed the capsule with sips of water and the tether was pulled back to image the oesophagus. SS-OCT at 1 000 000 A-scans/s enabled imaging 10 cm oesophageal lengths in 10 s with 30 µm transverse and 8 µm axial resolution. Capsule contact, longitudinal image coverage and patient toleration were assessed. RESULTS: Nine patients had non-dysplastic BE, three had ablative treatment-naïve neoplasia and four had prior ablation for dysplasia. Dry swallows facilitated capsule transit through the lower oesophageal sphincter (LES), and waiting 10 s before pullback reduced swallow induced LES relaxation. Slow nasal inhalation facilitated capsule retrieval and minimised gag reflex. The procedure was well tolerated. Ultrahigh-speed SS-OCT generated cross-sectional and subsurface en face images showing BE features, while subsurface en face images were required to assess the gastro-oesophageal junction. Candidate features of dysplasia were also identified which could inform follow-up endoscopy/biopsy. BE features were seen in all patients with histologically confirmed BE. Mean capsule contact over BE was 75%±27% for all patients and better in short segment BE. Mean longitudinal image coverage over BE was 59%±34% and better for long segment BE. CONCLUSIONS: Ultrahigh-speed tethered capsule SS-OCT can image en face and cross-sectional mucosal features over wide areas. Device and procedure optimisation improved performance. BE features could be identified in all patients, but limited capsule contact and longitudinal coverage could cause sampling errors for focal pathologies.

Quantitative Histopathology of Stained Tissues using Color Spatial Light Interference Microscopy (cSLIM).

Tissue biopsy evaluation in the clinic is in need of quantitative disease markers for diagnosis and, most importantly, prognosis. Among the new technologies, quantitative phase imaging (QPI) has demonstrated promise for histopathology because it reveals intrinsic tissue nanoarchitecture through the refractive index. However, a vast majority of past QPI investigations have relied on imaging unstained tissues, which disrupts the established specimen processing. Here we present color spatial light interference microscopy (cSLIM) as a new whole-slide imaging modality that performs interferometric imaging on stained tissue, with a color detector array. As a result, cSLIM yields in a single scan both the intrinsic tissue phase map and the standard color bright-field image, familiar to the pathologist. Our results on 196 breast cancer patients indicate that cSLIM can provide stain-independent prognostic information from the alignment of collagen fibers in the tumor microenvironment. The effects of staining on the tissue phase maps were corrected by a mathematical normalization. These characteristics are likely to reduce barriers to clinical translation for the new cSLIM technology.

Synthesis and Characterization of Heterobimetallic Iridium-Aluminum and Rhodium-Aluminum Complexes.

We demonstrate the synthesis and characterization of a new class of late-transition-metal-aluminum heterobimetallic complexes via a novel synthetic pathway. Complexes of this type are exceedingly rare. Joint experimental and theoretical data sheds light on the electronic effect of ligands containing aluminum moieties on late-transition-metal complexes.

Gradient light interference microscopy for 3D imaging of unlabeled specimens.

Multiple scattering limits the contrast in optical imaging of thick specimens. Here, we present gradient light interference microscopy (GLIM) to extract three-dimensional information from both thin and thick unlabeled specimens. GLIM exploits a special case of low-coherence interferometry to extract phase information from the specimen, which in turn can be used to measure cell mass, volume, surface area, and their evolutions in time. Because it combines multiple intensity images that correspond to controlled phase shifts between two interfering waves, gradient light interference microscopy is capable of suppressing the incoherent background due to multiple scattering. GLIM can potentially become a valuable tool for in vitro fertilization, where contrast agents and fluorophores may impact the viability of the embryo. Since GLIM is implemented as an add-on module to an existing inverted microscope, we anticipate that it will be adopted rapidly by the biological community.Challenges in biological imaging include labeling, photobleaching and phototoxicity, as well as light scattering. Here, Nguyen et al. develop a quantitative phase method that uses low-coherence interferometry for label-free 3D imaging in scattering tissue.

Automatic Gleason grading of prostate cancer using quantitative phase imaging and machine learning.

We present an approach for automatic diagnosis of tissue biopsies. Our methodology consists of a quantitative phase imaging tissue scanner and machine learning algorithms to process these data. We illustrate the performance by automatic Gleason grading of prostate specimens. The imaging system operates on the principle of interferometry and, as a result, reports on the nanoscale architecture of the unlabeled specimen. We use these data to train a random forest classifier to learn textural behaviors of prostate samples and classify each pixel in the image into different classes. Automatic diagnosis results were computed from the segmented regions. By combining morphological features with quantitative information from the glands and stroma, logistic regression was used to discriminate regions with Gleason grade 3 versus grade 4 cancer in prostatectomy tissue. The overall accuracy of this classification derived from a receiver operating curve was 82%, which is in the range of human error when interobserver variability is considered. We anticipate that our approach will provide a clinically objective and quantitative metric for Gleason grading, allowing us to corroborate results across instruments and laboratories and feed the computer algorithms for improved accuracy.

Halo-free Phase Contrast Microscopy.

We present a new approach for retrieving halo-free phase contrast microscopy (hfPC) images by upgrading the conventional PC microscope with an external interferometric module, which generates sufficient data for reversing the halo artifact. Acquiring four independent intensity images, our approach first measures haloed phase maps of the sample. We solve for the halo-free sample transmission function by using a physical model of the image formation under partial spatial coherence. Using this halo-free sample transmission, we can numerically generate artifact-free PC images. Furthermore, this transmission can be further used to obtain quantitative information about the sample, e.g., the thickness with known refractive indices, dry mass of live cells during their cycles. We tested our hfPC method on various control samples, e.g., beads, pillars and validated its potential for biological investigation by imaging live HeLa cells, red blood cells, and neurons.

Quantitative phase imaging of weakly scattering objects using partially coherent illumination.

In this paper, we extend our recent work on partially coherent quantitative phase imaging (pcQPI) from two-dimensional (2D) to three-dimensional (3D) imaging of weakly scattering samples. Due to the mathematical complexity, most theoretical modeling of quantitative phase image formation under partial coherence has focused on thin, well-focused samples. It is unclear how these abberations are affected by defocusing. Also, as 3D QPI techniques continue to develop, a better model needs to be developed to understand and quantify these aberrations when imaging thicker samples. Here, using the first order Born's approximation, we derived a mathematical framework that provides an intuitive model of image formation under varying degrees of coherence. Our description provides a clear connection between the halo effect and phase underestimation, defocusing and the 3D structure of the sample under investigation. Our results agree very well with the experiments and show that the microscope objective defines the sectioning ability of the imaging system while the condenser lens is responsible for the halo effect.

Plane-wave decomposition of spatially random fields.

We investigate the uniqueness of the plane-wave decomposition of temporally deterministic, spatially random fields. Specifically, we consider the decomposition of spatially ergodic and, thus, statistically homogeneous fields. We show that when the spatial power spectrum is injective, the plane waves are the only possible coherent modes. Furthermore, the randomness of such fields originates in the spatial spectral phase, i.e., the phase associated with the coefficients of each plane wave in the expansion. By contrast, the spectral amplitude is deterministic and is specified by the spatial power spectrum. We end with a discussion showing how the results can be translated in full to the time domain.

Deterministic signal associated with a random field.

Stochastic fields do not generally possess a Fourier transform. This makes the second-order statistics calculation very difficult, as it requires solving a fourth-order stochastic wave equation. This problem was alleviated by Wolf who introduced the coherent mode decomposition and, as a result, space-frequency statistics propagation of wide-sense stationary fields. In this paper we show that if, in addition to wide-sense stationarity, the fields are also wide-sense statistically homogeneous, then monochromatic plane waves can be used as an eigenfunction basis for the cross spectral density. Furthermore, the eigenvalue associated with a plane wave, exp[i(k · r-ωt)], is given by the spatiotemporal power spectrum evaluated at the frequency (k, ω). We show that the second-order statistics of these fields is fully described by the spatiotemporal power spectrum, a real, positive function. Thus, the second-order statistics can be efficiently propagated in the wavevector-frequency representation using a new framework of deterministic signals associated with random fields. Analogous to the complex analytic signal representation of a field, the deterministic signal is a mathematical construct meant to simplify calculations. Specifically, the deterministic signal associated with a random field is defined such that it has the identical autocorrelation as the actual random field. Calculations for propagating spatial and temporal correlations are simplified greatly because one only needs to solve a deterministic wave equation of second order. We illustrate the power of the wavevector-frequency representation with calculations of spatial coherence in the far zone of an incoherent source, as well as coherence effects induced by biological tissues.

Spatial Light Interference Microscopy (SLIM) using twisted-nematic liquid-crystal modulation.

We report the use of a twisted nematic liquid-crystal spatial light modulator (TNLC-SLM) for quantitative phase imaging. The experimental setup is a new implementation of the SLIM principle, which is a phase shifting, white light method for quantitative phase imaging. The approach is based on switching between the phase and amplitude modulation modes of the SLM. Our system is able to deliver a 0.99 nm spatial and 1.33 nm temporal pathlength sensitivity while retaining the optical transverse resolution. The system is implemented as an additional module mounted to a conventional microscope, which makes the system very easy to deploy and integrate with other imaging modalities.