Light-sheet microscopy: a tutorial

This paper is intended to give a comprehensive review of light-sheet (LS) microscopy from an optics perspective. As such, emphasis is placed on the advantages that LS microscope configurations present, given the degree of freedom gained by uncoupling the excitation and detection arms. The new imaging properties are first highlighted in terms of optical parameters and how these have enabled several biomedical applications. Then, the basics are presented for understanding how a LS microscope works. This is followed by a presentation of a tutorial for LS microscope designs, each working at different resolutions and for different applications. Then, based on a numerical Fourier analysis and given the multiple possibilities for generating the LS in the microscope (using Gaussian, Bessel, and Airy beams in the linear and nonlinear regimes), a systematic comparison of their optical performance is presented. Finally, based on advances in optics and photonics, the novel optical implementations possible in a LS microscope are highlighted.Peer ReviewedPostprint (published version

SKED: Sketch-guided Text-based 3D Editing

Text-to-image diffusion models are gradually introduced into computer graphics, recently enabling the development of Text-to-3D pipelines in an open domain. However, for interactive editing purposes, local manipulations of content through a simplistic textual interface can be arduous. Incorporating user guided sketches with Text-to-image pipelines offers users more intuitive control. Still, as state-of-the-art Text-to-3D pipelines rely on optimizing Neural Radiance Fields (NeRF) through gradients from arbitrary rendering views, conditioning on sketches is not straightforward. In this paper, we present SKED, a technique for editing 3D shapes represented by NeRFs. Our technique utilizes as few as two guiding sketches from different views to alter an existing neural field. The edited region respects the prompt semantics through a pre-trained diffusion model. To ensure the generated output adheres to the provided sketches, we propose novel loss functions to generate the desired edits while preserving the density and radiance of the base instance. We demonstrate the effectiveness of our proposed method through several qualitative and quantitative experiments

A model for adapting 3D graphics based on scalable coding, real-time simplification and remote rendering

Most current multiplayer 3D games can only be played on dedicated platforms, requiring specifically designed content and communication over a predefined network. To overcome these limitations, the OLGA (On-Line GAming) consortium has devised a framework to develop distributive, multiplayer 3D games. Scalability at the level of content, platforms and networks is exploited to achieve the best trade-offs between complexity and quality

Studies of the effects of gravitational and inertial forces on cardiovascular and respiratory dynamics

The current status and application are described of the biplane video roentgen densitometry, videometry and video digitization systems. These techniques were developed, and continue to be developed for studies of the effects of gravitational and inertial forces on cardiovascular and respiratory dynamics in intact animals and man. Progress is reported in the field of lung dynamics and three-dimensional reconstruction of the dynamic thoracic contents from roentgen video images. It is anticipated that these data will provide added insight into the role of shape and internal spatial relationships (which is altered particularly by acceleration and position of the body) of these organs as an indication of their functional status

Ex vivo live cell tracking in kidney organoids using light sheet fluorescence microscopy

Screening cells for their differentiation potential requires a combination of tissue culture models and imaging methods that allow for long-term tracking of the location and function of cells. Embryonic kidney re-aggregation in vitro assays have been established which allow for the monitoring of organotypic cell behaviour in re-aggregated and chimeric renal organoids. However, evaluation of cell integration is hampered by the high photonic load of standard fluorescence microscopy which poses challenges for imaging three-dimensional systems in real-time over a time course. Therefore, we employed light sheet microscopy, a technique that vastly reduces photobleaching and phototoxic effects. We have also developed a new method for culturing the re-aggregates which involves immersed culture, generating organoids which more closely reflect development in vivo. To facilitate imaging from various angles, we embedded the organoids in a freely rotatable hydrogel cylinder. Endpoint fixing and staining were performed to provide additional biomolecular information. We succeeded in imaging labelled cells within re-aggregated kidney organoids over 15 hours and tracking their fate while simultaneously monitoring the development of organotypic morphological structures. Our results show that Wt1-expressing embryonic kidney cells obtained from transgenic mice could integrate into re-aggregated chimeric kidney organoids and contribute to developing nephrons. Furthermore, the nascent proximal tubules that formed in the re-aggregated tissues using the new culture method displayed secretory function, as evidenced by their ability to secrete an organic anion mimic into the tubular lumen

3D time series analysis of cell shape using Laplacian approaches

Background: Fundamental cellular processes such as cell movement, division or food uptake critically depend on cells being able to change shape. Fast acquisition of three-dimensional image time series has now become possible, but we lack efficient tools for analysing shape deformations in order to understand the real three-dimensional nature of shape changes. Results: We present a framework for 3D+time cell shape analysis. The main contribution is three-fold: First, we develop a fast, automatic random walker method for cell segmentation. Second, a novel topology fixing method is proposed to fix segmented binary volumes without spherical topology. Third, we show that algorithms used for each individual step of the analysis pipeline (cell segmentation, topology fixing, spherical parameterization, and shape representation) are closely related to the Laplacian operator. The framework is applied to the shape analysis of neutrophil cells. Conclusions: The method we propose for cell segmentation is faster than the traditional random walker method or the level set method, and performs better on 3D time-series of neutrophil cells, which are comparatively noisy as stacks have to be acquired fast enough to account for cell motion. Our method for topology fixing outperforms the tools provided by SPHARM-MAT and SPHARM-PDM in terms of their successful fixing rates. The different tasks in the presented pipeline for 3D+time shape analysis of cells can be solved using Laplacian approaches, opening the possibility of eventually combining individual steps in order to speed up computations

MistForm: adaptive shape changing fog screens

We present MistForm, a shape changing fog display that can support one or two users interacting with either 2D or 3D content. Mistform combines affordances from both shape changing interfaces and mid-air displays. For example, a concave display can maintain content in comfortable reach for a single user, while a convex shape can support several users engaged on individual tasks. MistForm also enables unique interaction possibilities by exploiting the synergies between shape changing interfaces and mid-air fog displays. For instance, moving the screen will affect the brightness and blurriness of the screen at specific locations around the display, creating spaces with similar (collaboration) or different visibility (personalized content). We describe the design of MistForm and analyse its inherent challenges, such as image distortion and uneven brightness on dynamic curved surfaces. We provide a machine learning approach to characterize the shape of the screen and a rendering algorithm to remove aberrations. We finally explore novel interactive possibilities and reflect on their potential and limitations

Recent advances in optical tomography in low scattering media

Low scattering media is the best scenario for optical imaging in thick samples and deep tissue, as it allows to obtain high resolution images without suffering the limitations that the diffusion phenomenon imposes. The high contribution of ballistic light in this regime enabled the development of light sheet microscopy and optical projection tomography, two of the most common techniques nowadays in research laboratories. Their revolutionary approach and wide spectrum of applications and possibilities has lead to a frenetic rhythm of new works and techniques arising every year. The large amount of information available often overwhelms scientists and researchers trying to keep up to date with the last cutting edge advances of the field. This paper aims to give a brief review of the origins and fundamental aspects of these two techniques to focus on the most recent and yet non reviewed works. Apart from novel methods, this document also covers combined multimodal approaches and systems. To conclude, we put a spotlight on the important role that open-source microscopy systems play in the field, as they improve the accessibility to these techniques and promote collaborative networks across the optical imaging community