Automatic diagnosis and classification of breast surgical samples with dynamic full-field OCT and machine learning.

Purpose: The adoption of emerging imaging technologies in the medical community is often hampered when they provide a new unfamiliar contrast that requires experience to be interpreted. Dynamic full-field optical coherence tomography (D-FF-OCT) microscopy is such an emerging technique. It provides fast, high-resolution images of excised tissues with a contrast comparable to H&E histology but without any tissue preparation and alteration. Approach: We designed and compared two machine learning approaches to support interpretation of D-FF-OCT images of breast surgical specimens and thus provide tools to facilitate medical adoption. We conducted a pilot study on 51 breast lumpectomy and mastectomy surgical specimens and more than 1000 individual 1.3×1.3 mm2 images and compared with standard H&E histology diagnosis. Results: Using our automatic diagnosis algorithms, we obtained an accuracy above 88% at the image level (1.3×1.3 mm2) and above 96% at the specimen level (above cm2). Conclusions: Altogether, these results demonstrate the high potential of D-FF-OCT coupled to machine learning to provide a rapid, automatic, and accurate histopathology diagnosis with minimal sample alteration.

Full-field optical coherence tomography: novel imaging technique for extemporaneous high-resolution analysis of mucosal architecture in human gut biopsies.

Full-field optical coherence tomography (FFOCT) is an imaging technique of biological tissue based on tissue light reflectance analysis. We evaluated the feasibility of imaging fresh digestive mucosal biopsies after a quick mounting procedure (5 min) using two distinct modalities of FFOCT. In static FFOCT mode, we gained high-resolution images of general gut tissue-specific architecture, such as oesophageal papillae, gastric pits, duodenal villi and colonic crypts. In dynamic FFOCT mode, we imaged individual epithelial cells of the mucosal lining with a cellular or subcellular resolution and identified cellular components of the lamina propria. FFOCT represents a promising dye-free imaging tool for on-site analysis of gut tissue remodelling.

High-resolution handheld rigid endomicroscope based on full-field optical coherence tomography.

Full-field optical coherence tomography (FF-OCT) is a powerful tool for nondestructive assessment of biological tissue, i.e., for the structural examination of tissue in depth at a cellular resolution. Mostly known as a microscopy device for ex vivo analysis, FF-OCT has also been adapted to endoscopy setups since it shows good potential for in situ cancer diagnosis and biopsy guidance. Nevertheless, all the attempts to perform endoscopic FF-OCT imaging did not go beyond lab setups. We describe here, to the best of our knowledge, the first handheld FF-OCT endoscope based on a tandem interferometry assembly using incoherent illumination. A common-path passive imaging interferometer at the tip of an optical probe makes it robust and insensitive to environmental perturbations, and a low finesse Fabry-Perot processing interferometer guarantees a compact system. A good resolution (2.7 µm transverse and 6 µm axial) is maintained through the long distance, small diameter relay optics of the probe, and a good signal-to-noise ratio is achieved in a limited 100 ms acquisition time. High-resolution images and a movie of a rat brain slice have been recorded by moving the contact endoscope over the surface of the sample, allowing for tissue microscopic exploration at 20 m under the surface. These promising ex vivo results open new perspectives for in vivo imaging of biological tissue, in particular, in the field of cancer and surgical margin assessment.

Multi-modal acousto-optic/ultrasound imaging of ex vivo liver tumors at 790 nm using a Sn2 P2 S6 wavefront adaptive holographic setup.

Biological tissues are very strong light-scattering media. As a consequence, current medical imaging devices do not allow deep optical imaging unless invasive techniques are used. Acousto-optic imaging is a light-ultrasound coupling technique that takes advantage of the ballistic propagation of ultrasound in biological tissues to access optical contrast with a millimeter resolution. We have developed a photorefractive-crystal-based system that performs self-adaptive wavefront holography and works within the optical therapeutic window. As it works at an appropriate wavelength range for biological tissues imaging, it was tested on ex vivo liver samples containing tumors as a pre-clinical study. Optical contrast was obtained even if acoustical one was not significant. Ultrasound image (left) and acousto-optic image (right) of a liver biopsy with tumors. Acousto-optic imaging exhibits tumors that are not detected through ultrasound.

Flotillin microdomains stabilize cadherins at cell-cell junctions.

Cadherins are essential in many fundamental processes and assemble at regions of cell-cell contact in large macromolecular complexes named adherens junctions. We have identified flotillin 1 and 2 as new partners of the cadherin complexes. We show that flotillins are localised at cell-cell junctions (CCJs) in a cadherin-dependent manner. Flotillins and cadherins are constitutively associated at the plasma membrane and their colocalisation at CCJ increases with CCJ maturation. Using three-dimensional structured illumination super-resolution microscopy, we found that cadherin and flotillin complexes are associated with F-actin bundles at CCJs. The knockdown of flotillins dramatically affected N- and E-cadherin recruitment at CCJs in mesenchymal and epithelial cell types and perturbed CCJ integrity and functionality. Moreover, we determined that flotillins are required for cadherin association with GM1-containing plasma membrane microdomains. This allows p120 catenin binding to the cadherin complex and its stabilization at CCJs. Altogether, these data demonstrate that flotillin microdomains are required for cadherin stabilization at CCJs and for the formation of functional CCJs.

Dynamic ultrasound modulated optical tomography by self-referenced photorefractive holography.

Photorefractive Bi(12)SiO(20) single crystal is used for acousto-optic imaging in thick scattering media in the green part of the spectrum, in an adaptive speckle correlation configuration. Light fields at the output of the scattering sample exhibit typical speckle grains of 1 µm size within the volume of the nonlinear crystal. This heterogeneous illumination induces a complex refractive index structure without applying a reference beam on the crystal, leading to a self-referenced diffraction correlation scheme. We demonstrate that this simple and robust configuration is able to perform axially resolved ultrasound modulated optical tomography of thick scattering media with an improved optical etendue.

Acousto-optical coherence tomography with a digital holographic detection scheme.

Acousto-optical coherence tomography (AOCT) consists in using random phase jumps on ultrasound and light to achieve a millimeter resolution when imaging thick scattering media. We combined this technique with heterodyne off-axis digital holography. Two-dimensional images of absorbing objects embedded in scattering phantoms are obtained with a good signal-to-noise ratio. We study the impact of the phase modulation characteristics on the amplitude of the acousto-optic signal and on the contrast and apparent size of the absorbing inclusion.