Xuanwen Hua, Wenhao Liu, and Shu Jia, “High-resolution Fourier light-field microscopy for volumetric multi-color live-cell imaging,” Optica 8, 614 (2021)

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.

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

No more need to wait! Finally, DOIT 3D Micro is available! we are manufacturing the first MVP series to be delivered for early adopters. However, we are already accepting purchases on backorder and the units will be delivered in around one month.

We are overwhelmed with the nice feedback from many users. They did not think getting 3D information from their microscopic samples could be that easy and straightforward. Our plenoptic eyepiece is inserted into the ocular port (in the near future also in the trinocular) to replace the observer’s eye and transform that 2D image into a 3D multi perspectives views. This allows the user to get an “Extended Depth of Focus” (EDOF) of the sample immediately without the need for refocusing. Also, it allows seeing orthographic views to resolve angular information.

Doit 3D Micro makes 3D real-time microscopy easy!

Our system is so convenient for many applications. The user does not have to learn difficult processes or do the complex setups of other 3D microscopy techniques. Besides, most of the tools that the user applies habitually can be implemented too while using the DOIT 3D Micro. Another big advantage is sample handling and preparation. Most of the tools that get 3D information from the sample require delicate and long preparations. That increases substantially the risk of failure and therefore a huge waste of time and resources.

Real-time 3D microscope for all

One of the biggest advantages of the Plenoptic Eyepiece is 4D imaging. That means that the user is able to get information in the XYZ axis and also in real-time through the acquisition of videos. This is a huge leap for all those technicians and researchers who have dynamic events happening within their samples at a fast pace. Time-lapse imaging is not enough for them but the lightfield microscope they’ll get by attaching the DOIT 3D Micro will allow seeing the 4 dimensions. Just think about particle tracking velocimetry, neural activities monitoring, live-cell imaging reducing bleaching and phototoxicity, applying forces or chemicals to samples or materials to evaluate their response, etc… every movement within the microscopic sample can be acquired and displayed!

Reach us for more information! We will be very happy to understand which your 3D / 4D imaging needs and evaluate if we can help you by transforming your current microscope into a 3D lightfield microscope implementing the DOIT 3D Micro plenoptic eyepiece.

#LetsDOIT3D !

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

Miniscope3D system overview (From: Miniscope3D: optimized single-shot miniature 3D fluorescence microscopy)

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

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.

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