High-resolution Fourier light-field microscope

Researchers from the Georgia Institute of Technology and Emory University, extend the performances of Fourier lightfield concept by presenting the high-resolution Fourier light-field microscope (HR-FLM), which allows, among other applications, the fast and volumetric live-cell imaging.

Fourier lightfield microscopes (FLM) have the ability of capturing directly, in a single shot, a collection of orthographic perspective images, all with the same point spread function (PSF). Thus, deconvolution procedures are feasible and easily applicable. The authors of this paper take profit from these facts to go a step further in the limits achievable by of FLM.

To do this, the authors use a microscope objective with the highest NA ever used in FLM. Additionally, the microlens array is set in such a way that the aperture stop is fully covered by only three microlenses. These two facts give rise to perspective images with submicron resolution. Finally, an inverse computational process is implemented to retrieve the volume of the object through a wave-optics based Richardson–Lucy deconvolution of the perspective images and the 3D PSF.

Imaging mitochondria in fixed mammalian cells using HR-FLFM.

Authors show experiments that confirm that this FLM scheme allows to reconstruct the 3D image of sparse samples using a single camera frame, recovering a volume of 70µm x 10µm x 4µm, with lateral resolution of 0,5µm and axial resolution of 1,5 µm.

To conclude, authors anticipate HR-FLFM to offer a promising paradigm for interrogation of complex intracellular biomolecules, organelles, and microenvironments that underlie diverse spatiotemporal regulations of cellular processes and functions.

Commented by Dr. Manuel Martínez

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Miniscope3D: miniature fluorescence microscope

K. Yanny, N. Antipa, W. Liberti, S. Dehaeck, K. Monakhova, F. L. Liu, K. Shen, Ren Ng, and L. Waller, “Miniscope3D: optimized single-shot miniature 3D fluorescence microscopy,” Light Sci Appl. 9, 171 (2020). https://doi.org/10.1038/s41377-020-00403-7

Under the leadership of Laura Waller, researchers from the University of California, Berkeley, have reported a lightfield miniscope that is much smaller and lighter than previous ones, and that provides with 3D images with unprecedent resolution over a very large depth of field.

The 3D miniscope is based in the smart combination of three bright ideas: the Fourier lightfield concept, the use of an optimized multifocal phase mask, and the application of a rendering algorithm based on sparsity-constrained inverse methods

Based on those concepts, authors have built a demonstration prototype composed basically by a GRIN-lens objective, a phase mask inserted at the Fourier plane and a CMOS sensor placed at the mean focal length of the phase mask. With this prototype, authors have demonstrated the capability of render 3D images of sparse fluorescent samples with lateral resolution of  across a depth of 2,76µm across a depth of 390µm.

In this paper the Miniscope3D demonstrates its utility providing 3D images of mouse brain tissue and also of freely moving tardigrades. In summary, the miniscope3D provides single-shot 3D imaging for applications where a compact platform matters, such as volumetric neural imaging in freely moving animals and 3D motion studies of dynamic samples in incubators and lab-on-a-chip devices.

Commented by Dr. Manuel Martínez

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Plenoptic eyepiece. Transform any microscope into a 3D microscope

Doitplenotic will attend Focus on Microscopy 2021. Our scientific advisor Genaro Saavedra will present the paper titled “Plenoptic eyepiece. Transform any microscope into a 3D microscope”.

FOM2021 online will be the continuation of a yearly conference series on the latest innovations and developments in mostly optical microscopy and their application in biology, medicine, and the material sciences.

Key subjects for the conference series are the theory and practice of 3D optical imaging, related 3D image processing, and especially developments in resolution and imaging modalities. The conference series covers also the rapidly advancing fluorescence labeling techniques for confocal and multi-photon 3D imaging of -live- biological specimens.

Plenoptic eyepiece

Lightfield or plenoptic cameras are based on integral-photography concept and this can be applied to optical microscopy, but in the recent year a new architecture for lightfield microscopy has been proposed, it is named: Fourier lightfield, this scheme is based on collecting the spatio-angular field at the Fourier plane of the microscope [1].

The plenoptic eyepiece is based on Fourier lightfield, this is a portable, plug-and-play device that, after inserted at the ocular port, converts any conventional optical microscope into a lightfield microscope, with the best performance in terms of resolution and depth of field and allows you transform any microscope into a 3D microscope.

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Lightfield microscopy, a technique to study neural activities

Z. Zhang, L. Cong, L. Bai, and K. Wang, “Light-field microscopy for fast volumetric brain imaging,” Journal of Neuroscience Methods 352, 109083 (2021)

Researchers of the Center for Excellence in Brain Science and Intelligence Technology (Shanghai) have published this interesting paper in which a review of techniques for volumetric brain imaging is made.

Recording neural activities over large populations are critical for a better understanding of the functional mechanisms of animal brains. In this sense, the authors review different inspection techniques starting from those based on 3D scannings, like two-photon microscopy, which has the problem of low process speed and high light density. Another possibility, based on parallelizing the imaging process, is light-sheet microscopy which still has the problem requiring axial scanning.

Light-field-3d-microscopy for fast volumetric-brain-imaging

Confocal LFM (Zhang et al., 2020). MIPs over time of representative planes in reconstructed volumes in larval zebrafish brain (HuC: GCaMP6s). Scale bars, 50 μm.

In author’s opinion, lightfield microscopy (LFM) solves these problems elegantly by recording both the direction and location of light rays and achieving scanning-free and instantaneous volumetric imaging with a single camera exposure. However, this is made at the cost of a poor spatial resolution. But this drawback is overcome with the Fourier lightfield (FLFM) configuration, which shows substantially enhanced performance compared to that of conventional LFMs, including a lack of reconstruction artifacts near the focal plane, improved 3D reconstruction performance, and significantly reduced computational cost.

The paper finishes by collecting the results of LFM and FLFM when applied to brain images of different animals, like drosophila, zebrafish, or mouse. These results confirm the great utility of Fourier lightfield microscopy for brain imaging.

Commented by Dr. Manuel Martínez

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Doitplenoptic sponsors FOM 2021 Online Conference

The FOM conferences constitute an effective meeting point for developers and users working in this rapidly evolving field of microscopy and 3D imaging, playing an important role in the dissemination of information about new developments

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Light field microscopy session at SPAOM 2020

SPAOM 2020 will feature a full session on Light field microscopy. This session will take place, activities such as conferences, presentations, digital exhibitions, and workshops.

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SPAOM 2020 is co-organized and sponsored by DOITPLENOPTIC

SPAOM is the congress of Red Española de Microscopía Óptica Avanzada (REMOA) and Portuguese Platform of BioImagin (PPBI). Therefore, It aims to foster knowledge sharing within the community, covering current trends and new applications about optical imaging, sample preparation, or bioimage analysis.

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The 3D microscope for all is coming!

The first lightfield eyepiece for 3D microscopy

Great news for all microscopists! Our close-to-market prototype is almost ready! It’s been an exciting journey since BeAble Capital discovered the work that full professors Manuel Martínez-Corral and Genaro Saavedra together with our CTO Ángel Tolosa and other members of the 3DDisplayLab where developing.

Like many other breakthroughs, the idea behind it was simple but genius. Instruments with an ocular port are intended to be used by humans. But what if instead of a human eye, the observer is a machine?

A digital insect compound eye

Insect compound eye

See the microscopic world through the eyes of an insect

Similar to an insect compound eye, our plenoptic or lightfield technology uses microlenses to get different perspectives from the scene. As you might already know, 3D vision in our case is based on stereoscopic vision. We get a couple of images with slight parallax and our brain reconstructs the 3D scene.

Our eyepiece obtains multiple perspectives and the computer (the device’s brain) based on software algorithms, reconstructs the 3D microscopic information of the scene. This allows your conventional microscope to get 3D information from one ocular and 2D information from the other.

Convert your microscope into a 3D digital imaging microscope

As you can see in the picture below, the DOIT 3D Micro lightfield eyepiece is a cost-effective, easy-to-use, plug&play, and portable device that can be used in all microscopes provided with a standard ocular port.

Microscope 3D add-on

After the eyepiece is attached, the software process the optical information for 3D real-time image visualization and manipulation. Thanks to its different modules, the 3D image is presented in different ways that enhance diverse characteristics of the scene and its depth, such as views from several observation points, visualization of occlusions, selection of the focus plane along with the depth of field, an image with all planes focused at once, a depth map or topography of the surface of the inspected object, among others.

The system is very useful in many applications and is aimed at microscopy users who need high-quality 3D information from 1 micron (based on the native resolution of the microscope where it is coupled): researchers, technology centers, universities, R&D and inspection/QC departments, forensic work, …

Become a pioneer using the 3D lightfield microscope eyepiece!

Incredible, isn’t it? We know many microscopy users are tired of complex techniques and they just want to focus on their job instead of becoming physics experts. 3D microscopy solutions nowadays require many hours of training, bulky settings, and painful sample preparation, which increases the process times and the risk of undesired outcomes. And on top of that, big amounts of money investment!

We want to help scientists and technicians get rid of all of the above! We are now opening a pre-order period for a limited number of collaborators to do final testing on real application environments. If you have the right pioneer mindset, want to be the first of many, like to collaborate for the progress and easy access to science and, want to save some good money in the meantime… Join us!

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IRB Barcelona & Doitplenoptic collaboration

IRB Barcelona is “a world-class research center devoted to understanding fundamental questions about human health and disease. Among others, the Institute’s missions include conducting multidisciplinary research of excellence at the unique interface between biology, chemistry, and medicine, providing high-level training in the biomedical sciences to staff, students and visitors, driving innovation through active technology transfer to the benefit of society, and actively participating in an open dialogue with the public through a series of engagement and education activities.”

Doitplenoptic came across this incredible institute thanks to REMOA members during the 2019 SPAOM Meeting in Coimbra. Particularly when we met Julien Colombelli (Head of Advanced Digital Microscopy at IRB) one of the most reputed experts worldwide in advanced optical microscopy.

After explaining to him DOIT’s technology and our will of providing a 3D microscope for all, we were effortlessly on a good path for a collaboration agreement. Julien has this kind of fantastic scientific mindset. Always open to explore, improve, collaborate, and grow.

exceptional scientific results deserve to be transferred to society

Besides Julien’s disposition, technology transfer is key for IRB. As they claim “exceptional scientific results deserve to be transferred to society. With this in mind, IRB Barcelona has devised a proactive strategy to ensure that the discoveries made in its labs are developed into products and technologies that serve the scientific and healthcare communities, as well as society at large.”

These ingredients were the perfect receipt for the collaboration to happen: we agreed with IRB to conduct a pilot test project led by Julien Colombelli.

IRB Barcelona

Institut de Recerca Biomedica - Photo by IRB

This collaboration will help us to get closer to our mission of democratizing 3D microscopy. Julien and the rest of his team will detect any flaws, errors, or imperfections as well as suggest to us final improvements and modifications to deliver the best performance per each application where the 3D lightfield microscopy can make a substantial difference.

This sort of co-creation stage is a great leap for the final stages of development. Having the expert feedback of IRB adds great value both for the product itself and the go-to-market strategy before the official launch. We will understand better the needs of microscopists in different fields (life sciences, industry, clinical,…) looking for a 3D solution to perform their jobs.

Doitplenoptic is studying new proposals of collaboration similar to IRB’s pilot test. If you want to participate and help us bring a 3D microscope for all, drop us a line!

Let’s DOIT 3D!

PS: We also want to thank Tiago Oliveira from IRB who has made everything possible!

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