0000000000055995
AUTHOR
Gabriele Scrofani
Continuous Refocusing for Integral Microscopy with Fourier Plane Recording
Integral or light field imaging is an attractive approach in microscopy, as it allows to capture 3D samples in just one shot and explore them later through changing the focus on particular depth planes of interest. However, it requires a compromise between spatial and angular resolution on the 2D sensor recording the microscopic images. A particular setting called Fourier Integral Microscope (FIMic) allows maximizing the spatial resolution for the cost of reducing the angular one. In this work, we propose a technique, which aims at reconstructing the continuous light field from sparse FIMic measurements, thus providing the functionality of continuous refocus on any arbitrary depth plane. Ou…
Area-Based Depth Estimation for Monochromatic Feature-Sparse Orthographic Capture
With the rapid development of light field technology, depth estimation has been highlighted as one of the critical problems in the field, and a number of approaches have been proposed to extract the depth of the scene. However, depth estimation by stereo matching becomes difficult and unreliable when the captured images lack both color and feature information. In this paper, we propose a scheme that extracts robust depth from monochromatic, feature-sparse scenes recorded in orthographic sub-aperture images. Unlike approaches which rely on the rich color and texture information across the sub-aperture views, our approach is based on depth from focus techniques. First, we superimpose shifted …
Large Depth-of-Field Integral Microscopy by Use of a Liquid Lens
Integral microscopy is a 3D imaging technique that permits the recording of spatial and angular information of microscopic samples. From this information it is possible to calculate a collection of orthographic views with full parallax and to refocus computationally, at will, through the 3D specimen. An important drawback of integral microscopy, especially when dealing with thick samples, is the limited depth of field (DOF) of the perspective views. This imposes a significant limitation on the depth range of computationally refocused images. To overcome this problem, we propose here a new method that is based on the insertion, at the pupil plane of the microscope objective, of an electrical…
Fourier-domain lightfield microscopy: a new paradigm in 3D microscopy
Recently, integral (also known as lightfield or plenoptic) imaging concept has been applied successfully to microscopy. The main advantage of lightfield microscopy when compared with conventional 3D imaging techniques is that it offers the possibility of capturing the 3D information of the sample after a single shot. However, integral microscopy is now facing many challenges, like improving the resolution and depth of field of the reconstructed specimens or the development and optimization of specially-adapted reconstruction algorithms. This contribution is devoted to review a new paradigm in lightfield microscopy, namely, the Fourier-domain integral microscope (FiMic), that improves the ca…
Integral imaging with Fourier-plane recording
Integral Imaging is well known for its capability of recording both the spatial and the angular information of threedimensional (3D) scenes. Based on such an idea, the plenoptic concept has been developed in the past two decades, and therefore a new camera has been designed with the capacity of capturing the spatial-angular information with a single sensor and after a single shot. However, the classical plenoptic design presents two drawbacks, one is the oblique recording made by external microlenses. Other is loss of information due to diffraction effects. In this contribution report a change in the paradigm and propose the combination of telecentric architecture and Fourier-plane recordin…
Lightfield microscopy, an emerging tool for real-time 3D imaging
Integral, or lightfield, microscopy offers the possibility of capturing and processing in real time multiple views of 3D fluorescent samples captured with a single shot. In this contribution we review the recent advances in lightfield microscopy and enunciate the forthcoming challenges.
The Lightfield Microscope Eyepiece
Lightfield microscopy has raised growing interest in the last few years. Its ability to get three-dimensional information about the sample in a single shot makes it suitable for many applications in which time resolution is fundamental. In this paper we present a novel device, which is capable of converting any conventional microscope into a lightfield microscope. Based on the Fourier integral microscope concept, we designed the lightfield microscope eyepiece. This is coupled to the eyepiece port, to let the user exploit all the host microscope’s components (objective turret, illumination systems, translation stage, etc.) and get a 3D reconstruction of the sample. After the optical design, …
What about computational super-resolution in fluorescence Fourier light field microscopy?
Recently, Fourier light field microscopy was proposed to overcome the limitations in conventional light field microscopy by placing a micro-lens array at the aperture stop of the microscope objective instead of the image plane. In this way, a collection of orthographic views from different perspectives are directly captured. When inspecting fluorescent samples, the sensitivity and noise of the sensors are a major concern and large sensor pixels are required to cope with low-light conditions, which implies under-sampling issues. In this context, we analyze the sampling patterns in Fourier light field microscopy to understand to what extent computational super-resolution can be triggered duri…
Three-dimensional real-time darkfield imaging through Fourier lightfield microscopy
We report a protocol that takes advantage of the Fourier lightfield microscopy concept for providing 3D darkfield images of volumetric samples in a single-shot. This microscope takes advantage of the Fourier lightfield configuration, in which a lens array is placed at the Fourier plane of the microscope objective, providing a direct multiplexing of the spatio-angular information of the sample. Using the proper illumination beam, the system collects the light scattered by the sample while the background light is blocked out. This produces a set of orthographic perspective images with shifted spatial-frequency components that can be recombined to produce a 3D darkfield image. Applying the ade…
Robust Depth Estimation for Light Field Microscopy
Light field technologies have seen a rise in recent years and microscopy is a field where such technology has had a deep impact. The possibility to provide spatial and angular information at the same time and in a single shot brings several advantages and allows for new applications. A common goal in these applications is the calculation of a depth map to reconstruct the three-dimensional geometry of the scene. Many approaches are applicable, but most of them cannot achieve high accuracy because of the nature of such images: biological samples are usually poor in features and do not exhibit sharp colors like natural scene. Due to such conditions, standard approaches result in noisy depth ma…
FIMic: design for ultimate 3D-integral microscopy of in-vivo biological samples
In this work, Fourier integral microscope (FIMic), an ultimate design of 3D-integral microscopy, is presented. By placing a multiplexing microlens array at the aperture stop of the microscope objective of the host microscope, FIMic shows extended depth of field and enhanced lateral resolution in comparison with regular integral microscopy. As FIMic directly produces a set of orthographic views of the 3D-micrometer-sized sample, it is suitable for real-time imaging. Following regular integral-imaging reconstruction algorithms, a 2.75-fold enhanced depth of field and [Formula: see text]-time better spatial resolution in comparison with conventional integral microscopy is reported. Our claims …
3D Integral Microscopy based in far-field detection
Lately, Integral-Imaging systems have shown very promising capabilities of capturing the 3D structure of micro- scopic and macroscopic scenes. The aim of this work is to provide an optimal design for 3D-integral microscopy with extended depth of field and enhanced lateral resolution. By placing an array of microlenses at the aperture stop of the objective, this setup provides a set of orthographic views of the 3D sample. Adopting well known integral imaging reconstruction algorithms it can be shown that the depth of field as well as spatial resolution are improved with respect to conventional integral microscopy imaging. Our claims are supported on theoretical basis and experimental images …