Fourier Domain Optical Coherence Microscopy Research Articles

Extended focal depth Fourier domain optical coherence microscopy with a Bessel-beam - LP02 mode - from a higher order mode fiber.

We present a robust fiber-based setup for Bessel-like beam extended depth-of-focus Fourier-domain optical coherence microscopy, where the Bessel-like beam is generated in a higher order mode fiber module. In this module a stable guided LP02 core mode is selectively excited by a long period grating written in the higher order mode fiber. Imaging performance of this system in terms of lateral resolution and depth of focus was analyzed using samples of suspended microbeads and compared to the case where illumination is provided by the fundamental LP01 mode of a single mode fiber. Illumination with the LP02 mode allowed for a lateral resolution down to 2.5 µm as compared to 4.5 µm achieved with the LP01 mode of the single mode fiber. A three-fold enhancement of the depth of focus compared to a Gaussian beam with equally tight focus is achieved with the LP02 mode. Analysis of the theoretical lateral point spread functions for the case of LP01 and LP02 illumination agrees well with the experimental data. As the design space of waveguides and long-period gratings allows for further optimization of the beam parameters of the generated Bessel-like beams in an all-fiber module, this approach offers a robust and yet flexible alternative to free-space optics approaches or the use of conical fiber tips.

Numerically focused full-field swept-source optical coherence microscopy with structured illumination.

This paper presents an experimental investigation of the possibility of transverse resolution improvement combined with effective numerically focused 3D imaging in full-field swept-source optical coherence microscopy (OCM) by using structured illumination and specific numerical post-processing. The possibility of transverse resolution improvement of the OCM coherence signal combined with the possibility of numerical focusing is demonstrated by imaging a resolution test target in the optical focus and defocus regions. The possibility of numerically focused 3D imaging with high transverse resolution is further demonstrated by imaging a 3D phantom and a biological sample. The results obtained demonstrate the feasibility and prospects of the combination of structured illumination and numerical focusing in Fourier domain OCM.

Label-free fast 3D coherent imaging reveals pancreatic islet micro-vascularization and dynamic blood flow.

In diabetes, pancreatic β-cells play a key role. These cells are clustered within structures called islets of Langerhans inside the pancreas and produce insulin, which is directly secreted into the blood stream. The dense vascularization of islets of Langerhans is critical for maintaining a proper regulation of blood glucose homeostasis and is known to be affected from the early stage of diabetes. The deep localization of these islets inside the pancreas in the abdominal cavity renders their in vivo visualization a challenging task. A fast label-free imaging method with high spatial resolution is required to study the vascular network of islets of Langerhans. Based on these requirements, we developed a label-free and three-dimensional imaging method for observing islets of Langerhans using extended-focus Fourier domain Optical Coherence Microscopy (xfOCM). In addition to structural imaging, this system provides three-dimensional vascular network imaging and dynamic blood flow information within islets of Langerhans. We propose our method to deepen the understanding of the interconnection between diabetes and the evolution of the islet vascular network.

Volumetric imaging and quantification of cytoarchitecture and myeloarchitecture with intrinsic scattering contrast

We present volumetric imaging and computational techniques to quantify neuronal and myelin architecture with intrinsic scattering contrast. Using spectral / Fourier domain Optical Coherence Microscopy (OCM) and software focus-tracking we validate imaging of neuronal cytoarchitecture and demonstrate quantification in the rodent cortex in vivo. Additionally, by ex vivo imaging in conjunction with optical clearing techniques, we demonstrate that intrinsic scattering contrast is preserved in the brain, even after sacrifice and fixation. We volumetrically image cytoarchitecture and myeloarchitecture ex vivo across the entire depth of the rodent cortex. Cellular-level imaging up to the working distance of our objective (~3 mm) is demonstrated ex vivo. Architectonic features show the expected laminar characteristics; moreover, changes in contrast after the application of acetic acid suggest that entire neuronal cell bodies are responsible for the "negative contrast" present in the images. Clearing and imaging techniques that preserve tissue architectural integrity have the potential to enable non-invasive studies of the brain during development, disease, and remodeling, even in samples where exogenous labeling is impractical.

Numerical reconstruction of 3D image in Fourier domain confocal optical coherence microscopy

Along with high longitudinal resolution of optical coherence tomography, confocal optical coherence microscopy (OCM) provides high transversal resolution due to relatively high numerical apertures. However, the presence of relatively high numerical apertures leads to limited depth of field, which reduces the speed of OCM and decreases the advantage of Fourier domain detection. In this paper we propose a numerical processing technique for three-dimensional image reconstruction in Fourier domain OCM. It takes into account not only the defocus of different parts of imaged volumetric sample, but also the effects of upper layers’ refractive index on imaging the sample inner structure. Besides providing sharp coherence gated imaging, this technique also allows for determining both the geometrical thickness and refractive index of the sample layers.

Label-free imaging of cerebral β-amyloidosis with extended-focus optical coherence microscopy.

We demonstrate label-free imaging of cerebral β-amyloidosis ex vivo and in a living mouse model of Alzheimer's disease using extended-focus Fourier domain optical coherence microscopy (xfOCM). xfOCM provides 3D, high-resolution images of individual β-amyloid plaques in the brain parenchyma and vasculature and requires no staining of the alzheimeric sample under investigation. xfOCM also opens the possibility to perform minimally invasive studies of β-amyloid pathology in vivo, without the use of labeling methods, which potentially confound experimental findings.

High-resolution phase mapping with parallel Fourier domain optical coherence microscopy for dispersion contrast imaging

High-resolution phase mapping with parallel Fourier domain optical coherence microscopy for dispersion contrast imaging

Fiber-based combined optical coherence and multiphoton endomicroscopy

A fiber-based multimodal imaging system that combines multiphoton microscopy (MPM) with Fourier domain optical coherence microscopy (OCM) is reported. The system uses a fiber-based femtosecond laser, a fiber coupler, and a double-clad fiber (DCF) device. The fiber laser has a central wavelength of 1.04 μm and bandwidth of 29 nm. Longer excitation wavelength is used to increase penetration depth and increase the excitation efficiency for dyes, such as red fluorescent dyes. A single mode fiber coupler is used to replace the free-space beam splitter and one arm of the coupler is fused with a double-clad fiber device. The MPM and OCM share the same excitation light path in the core of a double-clad fiber, while the OCM and MPM signals were collected by the core and clad of the double-clad fiber, respectively. The performance of the introduced double-clad device is analyzed. The device can confine all the excitation light in the core and has a collection efficiency of 20% for the MPM signal. The efficiency can be further increased by fusing more multimode fibers with the DCF. Simultaneous optical coherence microscopic imaging, second harmonic generation imaging, and two-photon excitation fluorescence imaging are demonstrated in biological samples.

Dark-field optical coherence microscopy

Dark-field illumination is known to enhance scattering contrast in optical microscopy. We combined this concept with Fourier domain optical coherence microscopy (OCM). The detection and illumination paths are decoupled, and only the scattered light originating from the sample generates the tomogram signal, whereas any specular reflection is highly suppressed. We analyze and discuss this dark-field OCM concept and present its superior imaging quality on live cell samples.

Extended focus depth for Fourier domain optical coherence microscopy

We report on a new detection scheme for Fourier domain optical coherence microscopy that exhibits high transverse resolution along an axially extended focal range. Nearly constant transverse resolution of approximately 1.5 microm along a focal range of 200 microm is experimentally verified with a maximum sensitivity of 105 dB. A broad-bandwidth Ti:sapphire laser allowed for an axial resolution of 3 microm in air.