arXiv subjects

Kate Grieve

Publications and source records attributed to Kate Grieve.

14 recordsLinked to original sources

arXiv2025

Rolling phase modulation regime for dynamic full field OCT

Dynamic full-field optical coherence tomography (DFFOCT) has recently emerged as an invaluable label-free microscopy technique, owing to its sensitivity to cell activity, as well as speed and sectioning ability. However, the quality of DFFOCT images are often degraded due to phase noise and fringe artifacts. In this work, we present a new implementation named Rolling-Phase (RP) DFFOCT, in which the reference arm is slowly scanned over magnitudes exceeding 2$\pi$. We demonstrate mathematically and experimentally that it shows superior image quality while enabling to extract both static and dynamic contrast simultaneously. We showcase RP DFFOCT on monkey retinal explant, and demonstrate its ability to better resolve subcellular structures, including intranuclear activity.

q-bio.QM↗

arXiv2023

Interface Self-Referenced Dynamic Full-Field Optical Coherence Tomography

Dynamic full-field optical coherence tomography (D-FFOCT) has recently emerged as an invaluable live label-free and non-invasive imaging modality able to image subcellular biological structures and their metabolic activity within complex 3D samples. However, D-FFOCT suffers from fringe artefacts when imaging nearby reflective surfaces and is highly sensitive to vibrations. Here, we present interface Self-Referenced (iSR) D-FFOCT, an alternative configuration to D-FFOCT that takes advantage of the presence of the sample coverslip in between the sample and the objective by using it as a defocused reference arm, thus avoiding the aforementioned artefacts. We demonstrate the ability of iSR D-FFOCT to image 2D fibroblast cell cultures, which are among the flattest mammalian cells.

physics.optics↗

arXiv2023

Dynamic Full-Field Optical Coherence Tomography module adapted to commercial microscopes for longitudinal in vitro cell culture study

Dynamic full-field optical coherence tomography (D-FFOCT) has recently emerged as a label-free imaging tool, capable of resolving cell types and organelles within 3D live samples, whilst monitoring their activity at tens of milliseconds resolution. Here, a D-FFOCT module design is presented which can be coupled to a commercial microscope with a stage top incubator, allowing non-invasive label-free longitudinal imaging over periods of minutes to weeks on the same sample. Long term volumetric imaging on human induced pluripotent stem cell-derived retinal organoids is demonstrated, highlighting tissue and cell organisation as well as cell shape, motility and division. Imaging on retinal explants highlights single 3D cone and rod structures. An optimal workflow for data acquisition, postprocessing and saving is demonstrated, resulting in a time gain factor of 10 compared to prior state of the art. Finally, a method to increase D-FFOCT signal-to-noise ratio is demonstrated, allowing rapid organoid screening.

q-bio.QM↗

arXiv2022

Compact orthogonal view OCT: clinical exploration of human trabecular meshwork and cornea at cell resolution

Glaucoma remains among the leading causes of blindness despite many treatment options available today. Effective treatment requires early diagnosis, which is difficult to achieve with existing imaging technologies that detect the already inflicted irreversible posterior eye damage. One improved approach could be to detect the changes in trabecular meshwork of the anterior eye that precede the intraocular pressure buildup and vision loss. Here we report a device that enables in vivo exploration of trabecular meshwork at cell resolution. The instrument combines two orthogonal views: en face view of time-domain full-field OCT for high-resolution and cross-sectional view of spectral-domain OCT for aligning to the meshwork. The device resolves micron-level corneoscleral and uveal pores, meshwork beams and internal elastic fibers, ciliary body tendons and trabecular cell nuclei. Capable of detecting blockage of trabecular pores, orthogonal view OCT presents a promising solution to the long-standing problem of early glaucoma diagnosis. Future clinical adoption of the device is facilitated by the compact footprint and broad functionality that includes corneal imaging in healthy subjects and patients (with keratoconus, Fuchs endothelial dystrophy) using the same design.

physics.med-ph↗

arXiv2021

Optical phase modulation by natural eye movements: application to time-domain FF-OCT image retrieval

Eye movements are commonly seen as an obstacle to high-resolution ophthalmic imaging. In this context we study the natural axial movements of the in vivo human eye and show that they can be used to modulate the optical phase and retrieve tomographic images via time-domain full-field optical coherence tomography (TD-FF-OCT). This approach opens a path to a simplified ophthalmic TD-FF-OCT device, operating without the usual piezo motor-camera synchronization. The device demonstrates in vivo human corneal images under the different image retrieval schemes (2-phase and 4-phase) and different exposure times (3.5 ms, 10 ms, 20 ms). Data on eye movements, acquired with a 100 kHz spectral-domain OCT with axial eye tracking, are used to study the influence of ocular motion on the probability of capturing high-signal tomographic images without phase washout. The optimal combinations of camera acquisition speed and amplitude of piezo modulation are proposed and discussed.

physics.med-ph↗

arXiv2021

Non invasive live imaging of a novel retinal pigment epithelium stress model with Dynamic Full-Field Optical Coherence Tomography

Retinal degenerative diseases lead to the blindness of millions of people around the world. In case of age-related macular degeneration (AMD), the atrophy of retinal pigment epithelium (RPE) precedes neural dystrophy. But as crucial as understanding both healthy and pathological RPE cell physiology is for those diseases, no current technique allows subcellular in vivo or in vitro live observation of this critical cell layer. To fill this gap, we propose dynamic full-field OCT (D-FFOCT) as a candidate for live observation of in vitro RPE phenotype. In this way, we monitored primary porcine and human stem cell-derived RPE cells in stress model conditions by performing scratch assays. In this study, we quantified wound healing parameters on the stressed RPE, and observed different cell phenotypes, displayed by the D-FFOCT signal. In order to decipher the subcellular contributions to these dynamic profiles, we performed immunohistochemistry to identify which organelles generate the signal and found mitochondria to be the main contributor to D-FFOCT contrast. Altogether, D-FFOCT appears to be an innovative method to follow degenerative disease evolution and could be an appreciated method in the future for live patient diagnostics and to direct treatment choice.

physics.bio-ph↗

arXiv2020

Super-resolution in vivo retinal imaging using structured illumination ophthalmoscopy

Structured illumination microscopy (SIM) is one of the most versatile super-resolution techniques. Yet, its application to live imaging has been so far mainly limited to fluorescent and stationary specimens. Here, we present advancements in SIM to jointly tackle all the challenges of imaging living samples, i.e., obtaining super-resolution over an undistorted wide-field while dealing with sample motion, scattering, sample-induced optical aberrations and low signal-to-noise ratio. By using adaptive optics to compensate for optical aberrations and a reconstruction algorithm tailored for a moving and thick tissue, we successfully applied SIM to in vivo retinal imaging and demonstrated structured illumination ophthalmoscopy for high contrast super-resolution in vivo imaging of the human retina.

physics.med-ph↗

arXiv2020

Adaptive-glasses wavefront sensorless full-field OCT for high-resolution retinal imaging over a wide field-of-view

The highest three-dimensional (3D) resolution possible in in-vivo retinal imaging is achieved by combining optical coherence tomography (OCT) and adaptive optics (AO). However, this combination brings important limitations, such as small field-of-view and complex, cumbersome systems, preventing so far the translation of this technology from the research lab to clinics. In this Letter, we introduce an approach that avoids these limitations by using a multi-actuator adaptive lens just in front of the eye, in a technique we call the adaptive-glasses wavefront sensorless approach. We implemented this approach on our compact full-field OCT (FFOCT) retinal imager without increasing its footprint or optical complexity. The correction of ocular aberrations through the adaptive-glasses approach increased the FFOCT signal-to-noise ratio and enabled us to image different retinal layers with a 3D cellular resolution in a 5{\deg} $\times$ 5{\deg} field-of-view, without apparent anisoplanatism.

physics.med-ph↗

arXiv2020

Optical Incoherence Tomography: a method to generate tomographic retinal cross-sections with non-interferometric imaging systems

Optical tomographic cross-sectional images of biological samples were made possible by interferometric imaging techniques such as Optical Coherence Tomography (OCT). Owing to its unprecedented view of the sample, OCT has become a gold standard, namely for human retinal imaging in the clinical environment. In this Letter, we present Optical Incoherence Tomography (OIT): a completely digital method extending the possibility to generate tomographic retinal cross-sections to non-interferometric imaging systems such as en-face AO-ophthalmoscopes. We demonstrate that OIT can be applied to different imaging modalities using back-scattered and multiply-scattered light including systems without inherent optical sectioning. We show that OIT can be further used to guide focus position when the user is "blind" focusing, allowing precise imaging of translucent retinal structures, the vascular plexuses and the retinal pigment epithelium using respectively split detection, motion contrast, and autofluorescence techniques.

physics.med-ph↗

arXiv2020

Real-time, non-contact, cellular imaging and angiography of human cornea and limbus with common-path Full-field/SDOCT

In today's clinics, a cellular-resolution view of the cornea can be achieved only with an in vivo confocal microscope (IVCM) in contact with the eye. Here, we present a common-path Full-field/Spectral-domain OCT microscope (FF/SD OCT), which, for the first time, enables cell-detail imaging of the entire ocular surface (central and peripheral cornea, limbus, sclera, tear film) without contact and in real time. The device, which has been successfully tested in human subjects, is now ready for direct implementation in clinical research. Real-time performance is achieved through rapid axial eye tracking and simultaneous defocusing correction. Images, extracted from real-time videos, contain cells and nerves, which can be quantified over a millimetric field-of-view, beyond the capability of IVCM and conventional OCT. In the limbus, Palisades of Vogt, vessels and blood flow can be resolved with high contrast without contrast agent injection. The fast imaging speed of 275 frames/s (0.6 billion pixels/s) allowed direct monitoring of blood flow dynamics, enabling creation of high-resolution velocity maps for the first time. Tear flow velocity and evaporation time could be measured without fluorescein administration.

physics.med-ph↗

arXiv2020

Curved-Full-Field OCT for high-resolution imaging of living human retina over a large field-of-view

Allying high-resolution with a large field-of-view (FOV) is of great importance in the fields of biology and medicine, but particularly challenging when imaging non-flat living samples such as the human retina. Indeed, high-resolution is normally achieved with adaptive optics (AO) and scanning methods, which considerably reduce the useful FOV and increase the system complexity. An alternative technique is time-domain Full-Field Optical Coherence Tomography (FF-OCT), which has already shown its potential for in-vivo high-resolution retinal imaging. Here, we introduce curved-field FF-OCT, capable of matching the coherence gate geometry to the sample curvature, thus achieving a larger FOV than previously possible. Using this instrument, we obtained high-resolution images of living human photoreceptors close to the foveal center without AO and with a 1 mm x 1 mm FOV in a single shot. This novel advance enables the extraction of photoreceptor-based biomarkers with ease and spatiotemporal monitoring of individual photoreceptors. We compare our findings with AO-assisted ophthalmoscopes, highlighting the potential of FF-OCT, as a relatively simple, compact and low-cost system, to become a routine clinical imaging technique.

physics.med-ph↗

arXiv2019

Dynamic full-field optical coherence tomography: 3D live-imaging of retinal organoids

Optical coherence tomography offers astounding opportunities to image the complex structure of living tissue, but lacks functional information. We present dynamic full-field optical coherence tomography to image living human induced pluripotent stem cell-derived retinal organoids non-invasively. Colored images with an endogenous contrast linked to organelle motility are generated, with sub-micrometer spatial resolution and millisecond temporal resolution, opening an avenue to identify specific cell types in living tissue via their function.

physics.bio-ph↗

arXiv2018

In vivo high resolution human retinal imaging with wavefront correctionless full-field OCT

As the lateral resolution of FFOCT with spatially incoherent illumination has been shown to be insensitive to aberrations, we demonstrate high resolution en face full-field OCT (FFOCT) retinal imaging without wavefront correction in the human eye in vivo for the first time. A combination of FFOCT with spectral-domain OCT (SDOCT) is applied for real-time matching of the optical path lengths (OPL) of FFOCT. Through the real-time cross-sectional SDOCT images, the OPL of the FFOCT reference arm is matched with different retinal layers in the FFOCT sample arm. Thus, diffraction limited FFOCT images of multiple retinal layers are acquired at both the near periphery and the fovea. The en face FFOCT retinal images reveal information about various structures such as the nerve fiber orientation, the blood vessel distribution, and the photoreceptor mosaic.

physics.med-ph↗

arXiv2013

Imaging of non tumorous and tumorous human brain tissue with full-field optical coherence tomography

A prospective study was performed on neurosurgical samples from 18 patients to evaluate the use of Full-Field Optical Coherence Tomography (FF-OCT) in brain tumor diagnosis. FF-OCT captures en face slices of tissue samples at 1\mum resolution in 3D with a typical 200\mum imaging depth. A 1cm2 specimen is scanned at a single depth and processed in about 5 minutes. This rapid imaging process is non-invasive and 30 requires neither contrast agent injection nor tissue preparation, which makes it particularly well suited to medical imaging applications. Temporal chronic epileptic parenchyma and brain tumors such as meningiomas, low- grade and high-grade gliomas, and choroid plexus papilloma were imaged. A subpopulation of neurons, myelin fibers and CNS vasculature were clearly identified. Cortex could be discriminated from white matter, but individual glial cells as astrocytes (normal or reactive) or oligodendrocytes were not observable. This study reports for the first time on the feasibility of using FF-OCT in a real-time manner as a label-free non-invasive imaging technique in an intra-operative neurosurgical clinical setting to assess tumorous glial and epileptic margins.

physics.med-ph↗