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At least 19 records

REMBI: Recommended Metadata for Biological Images—enabling reuse of microscopy data in biology

Bioimaging data have significant potential for reuse, but unlocking this potential requires systematic archiving of data and metadata in public databases. Here, we propose draft metadata guidelines to begin addressing the needs of diverse communities within light and electron microscopy. We hope this publication and the proposed Recommended Metadata for Biological Images (REMBI) will stimulate discussions about their implementation and future extension.

59 BASIC BIOLOGICAL SCIENCES↗

Biological Imaging Capability in the ABRS Facility on ISS

This slide presentation reviews the Advanced Biological Research System (ABRS) on the International Space Station (ISS) and its biological imaging capability. The ABRS is an environmental control chamber. It has two indpendently controlled Experiment Research Chambers (ERCs) with temperature, relative humidity and carbon dioxide controls. ABRS is a third generation plant growth system. Several experiments are reviewed, with particular interest in the use of Green Fluorescent Protein (GFP) a non-destructive plant stress reporting mechanism, naturally found in jellyfish.

Cox, David R.↗

Broadband coherent anti-Stokes Raman scattering (BCARS) microscopy for rapid, label-free biological imaging

Broadband coherent anti-Stokes Raman scattering (BCARS) microscopy is a label-free imaging approach that provides detailed chemical information at high spatial resolution in a sample through nonlinear, coherent excitation of molecular vibrations and detection of Raman spectra. While its utility for biological imaging has been demonstrated, many aspects of this technique must mature before it can be widely adopted. One of the areas of required improvement is imaging speed—most BCARS implementations involve sample rastering, which limits imaging speed. Beam scanning can provide faster BCARS imaging but presents some unique challenges. Here, we describe a beam-scanning BCARS microscopy system that improves spatial resolution twofold and imaging speed by fivefold over a previous beam-scanning implementation. These enhancements were enabled by an improvement in supercontinuum power and the use of a sCMOS camera for its high data transfer rate and low read noise. Implementation of the sCMOS camera required correction for the significant pixel-to-pixel background and photon response nonuniformity. Here, we report on the method that we implemented for calibrating and correcting the pixel-to-pixel differences in sCMOS camera noise.

Dixon, Jessica Z. [Georgia Institute of Technology↗

Photocaged functional nucleic acids for spatiotemporal imaging in biology

Imaging of species in living organisms with high spatiotemporal resolution is essential for understanding biological processes. While functional nucleic acids (FNAs), such as catalytic nucleic acids and aptamers, have emerged as effective sensors for a wide range of molecules, photocaged control of these FNAs has played a key role in translating them into bioimaging agents with high spatiotemporal control. In this paper, we summarize methods and results of photocaged FNAs based on photolabile modifications, photoisomerization, and photothermal activation. Future directions, including strategies to improve the performance of these photocaged FNAs, are also described.

59 BASIC BIOLOGICAL SCIENCES↗

VLM4Bio: A Benchmark Dataset to Evaluate Pretrained Vision-Language Models for Trait Discovery from Biological Images

Images are increasingly becoming the currency for documenting biodiversity on the planet, providing novel opportunities for accelerating scientific discoveries in the field of organismal biology, especially with the advent of large vision-language models (VLMs). We ask if pre-trained VLMs can aid scientists in answering a range of biologically relevant questions without any additional fine-tuning. In this paper, we evaluate the effectiveness of 12 state-of-the-art (SOTA) VLMs in the field of organismal biology using a novel dataset, VLM4Bio, consisting of 469K question8 answer pairs involving 30K images from three groups of organisms: fishes, birds, and butterflies, covering five biologically relevant tasks. We also explore the effects of applying prompting techniques and tests for reasoning hallucination on the performance of VLMs, shedding new light on the capabilities of current SOTA VLMs in answering biologically relevant questions using images

Maruf, M [Virginia Tech, Blacksburg]↗

Biological imaging spectroscopy

Improved detectors, new electro-optical devices and vastly improved computational power for data analysis have fueled the recent interest in combining biology and spectroscopy.

biophotonics imaging spectroscopy↗

Method for imaging informational biological molecules on a semiconductor substrate

Imaging biological molecules such as DNA at rates several times faster than conventional imaging techniques is carried out using a patterned silicon wafer having nano-machined grooves which hold individual molecular strands and periodically spaced unique bar codes permitting repeatably locating all images. The strands are coaxed into the grooves preferably using gravity and pulsed electric fields which induce electric charge attraction to the molecular strands in the bottom surfaces of the grooves. Differential imaging removes substrate artifacts.

Coles, L. Stephen↗

Spectrum – An Instrument for Multispectral Biological Fluorescence Imaging on the International Space Station

Spectrum is a compact instrument for fluorescent imaging of biological specimens in microgravity and capable of operating within the constraints of an EXPRESS Rack on the International Space Station (ISS). The Spectrum Flight Unit (SFU) was launched to ISSin November 2019 and is available for science investigations as a general instrument. A duplicate Engineering Development Unit (EDU) is available at NASA Kennedy Space Center (KSC). Science Verification Testing (SVT) performed at KSCdemonstrated all required functions including five (5) fluorescence excitation wavelengths, growth lighting, environmental controls and monitoring, and high-resolution imaging using a 71 Megapixel camera with flat field lens. The design features key Orbital Replaceable Unit (ORU) components, which may be adapted or changed by future investigators for tailored science experiments.

Bradley Burns↗

In Liquid Infrared Scattering Scanning Near-Field Optical Microscopy for Chemical and Biological Nanoimaging

Imaging biological systems with simultaneous intrinsic chemical specificity and nanometer spatial resolution in their typical native liquid environment has remained a long-standing challenge. In this paper we demonstrate a general approach of chemical nanoimaging in liquid based on infrared scattering scanning near-field optical microscopy (IR $s$-SNOM). It is enabled by combining AFM operation in a fluid cell with evanescent IR illumination via total internal reflection, which provides spatially confined excitation for minimized IR water absorption, reduced far-field background, and enhanced directional signal emission and sensitivity. We demonstrate in-liquid IR $s$-SNOM vibrational nanoimaging and conformational identification of catalase nano-crystals and spatio-spectral analysis of biomimetic peptoid sheets with monolayer sensitivity and chemical specificity at the few zeptomole level. This work establishes the principles of in-liquid and in-situ IR $s$-SNOM spectroscopic chemical nano-imaging and its general applicability to biomolecular, cellular, catalytic, electrochemical, or other interfaces and nano-systems in liquids or solutions.

77 NANOSCIENCE AND NANOTECHNOLOGY↗

Toward implementing autonomous adaptive data acquisition for scanning hyperspectral imaging of biological systems

Autonomous experimentation is an emerging area of research, primarily related to autonomous vehicles, scientific combinatorial discovery approaches in materials science and drug discovery, and iterative research loops of planning, experimentation, and analysis. However, autonomous approaches developed in these contexts are difficult to apply to high-dimensional mapping technologies, such as scanning hyperspectral imaging of biological systems, due to sample complexity and heterogeneity. We briefly cover the history of adaptive sampling algorithms and surrogate modeling in order to define autonomous adaptive data acquisition as an objective-based, flexible building block for future biological imaging experimentation driven by intelligent infrastructure. We subsequently summarize the recent implementations of autonomous adaptive data acquisition (AADA) for scanning hyperspectral imaging, assess how these address the difficulties of autonomous approaches in hyperspectral imaging, and highlight the AADA design variation from a goal-oriented perspective. Finally, we present a modular AADA architecture that embeds AADA-driven flexible building blocks to address the challenge of time resolution for high-dimensional scanning hyperspectral imaging of nonequilibrium dynamical systems. In our example research-driven experimental design case, we propose an AADA infrastructure for time-resolved, noninvasive, and label-free scanning hyperspectral imaging of living biological systems. This AADA infrastructure can accurately target the correct state of the system for experimental workflows that utilize subsequent expensive, high-information-content analytical techniques.

59 BASIC BIOLOGICAL SCIENCES↗

Three-Dimensional Mass Spectrometric Imaging of Biological Structures Using a Vacuum-Compatible Microfluidic Device

Three-dimensional (3D) molecular imaging of biological structures is important for a wide range of research. In recent decades, secondary ion mass spectrometry (SIMS) has been recognized as a powerful technique for both two-dimensional (2D) and 3D molecular imaging. Sample fixations (e. g., chemical fixation and cryogenic fixation methods) are necessary to adapt biological samples to the vacuum condition in the SIMS chamber, which has been demonstrated to be non-trivial and less controllable, thus limiting the wider application of SIMS on 3D molecular analysis of biological samples. Our group recently developed in situ liquid SIMS that offers great opportunities for the molecular study of various liquids and liquid interfaces. In this work, we demonstrate that a further development of the vacuum-compatible microfluidic device used in in situ liquid SIMS provides a convenient freeze-fixation of biological samples and leads to more controllable and convenient 3D molecular imaging. The special design of this new vacuum-compatible liquid chamber allows an easy determination of sputter rates of ice, which is critical for calibrating the depth scale of frozen biological samples. Sputter yield of a 20 keV Ar 1800 + ion on ice has been determined as 1500 (± 8%) water molecules per Ar 1800 + ion, consistent with our results from molecular dynamics simulations. Moreover, using the information of ice sputter yield, we successfully conduct 3D molecular imaging of frozen homogenized milk and observe network structures of interesting organic and inorganic species. Finally, taken together, our results will significantly benefit various research fields relying on 3D molecular imaging of biological structures.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Biological Visualization, Imaging and Simulation(Bio-VIS) at NASA Ames Research Center: Developing New Software and Technology for Astronaut Training and Biology Research in Space

The Bio- Visualization, Imaging and Simulation (BioVIS) Technology Center at NASA's Ames Research Center is dedicated to developing and applying advanced visualization, computation and simulation technologies to support NASA Space Life Sciences research and the objectives of the Fundamental Biology Program. Research ranges from high resolution 3D cell imaging and structure analysis, virtual environment simulation of fine sensory-motor tasks, computational neuroscience and biophysics to biomedical/clinical applications. Computer simulation research focuses on the development of advanced computational tools for astronaut training and education. Virtual Reality (VR) and Virtual Environment (VE) simulation systems have become important training tools in many fields from flight simulation to, more recently, surgical simulation. The type and quality of training provided by these computer-based tools ranges widely, but the value of real-time VE computer simulation as a method of preparing individuals for real-world tasks is well established. Astronauts routinely use VE systems for various training tasks, including Space Shuttle landings, robot arm manipulations and extravehicular activities (space walks). Currently, there are no VE systems to train astronauts for basic and applied research experiments which are an important part of many missions. The Virtual Glovebox (VGX) is a prototype VE system for real-time physically-based simulation of the Life Sciences Glovebox where astronauts will perform many complex tasks supporting research experiments aboard the International Space Station. The VGX consists of a physical display system utilizing duel LCD projectors and circular polarization to produce a desktop-sized 3D virtual workspace. Physically-based modeling tools (Arachi Inc.) provide real-time collision detection, rigid body dynamics, physical properties and force-based controls for objects. The human-computer interface consists of two magnetic tracking devices (Ascention Inc.) attached to instrumented gloves (Immersion Inc.) which co-locate the user's hands with hand/forearm representations in the virtual workspace. Force-feedback is possible in a work volume defined by a Phantom Desktop device (SensAble inc.). Graphics are written in OpenGL. The system runs on a 2.2 GHz Pentium 4 PC. The prototype VGX provides astronauts and support personnel with a real-time physically-based VE system to simulate basic research tasks both on Earth and in the microgravity of Space. The immersive virtual environment of the VGX also makes it a useful tool for virtual engineering applications including CAD development, procedure design and simulation of human-system systems in a desktop-sized work volume.

Smith, Jeffrey↗

Cryogenic electron tomography by the numbers: Charting underexplored lineages in structural cell biology

Imaging cells and their interactions across the whole biosphere with molecular-scale resolution is key for understanding structure–function relations. Cryogenic electron tomography (cryo-ET) is a powerful method for obtaining this critical information. However, cryo-ET studies are challenging and often limited to a small number of cell types per study. Here, we collate cryo-ET data from hundreds of cells and tissues across the biosphere to i) identify emerging methodological trends, ii) pinpoint strategies to reduce imaging time and costs, iii) quantitatively compare methods for cell freezing and sectioning, and iv) census cryo-ET species coverage across all domains of life. Comparing the fraction of cellular material within a single lamella across all domains of life reveals an order of magnitude difference between eukaryotes (1%) compared to bacteria (9%) and archaea (14%). We calculate the fraction of cellular material which can be imaged using distinct sectioning methods on multicellular communities and tissues—identifying serial lift-out as a powerful approach for obtaining more complete cellular depictions. Finally, we show that the biodiversity of current cryo-ET studies is 2 to 3 orders of magnitude lower than in sequence libraries and 4 to 5 lower than the total predicted on Earth. Our analyses reveal major evolutionary lineages which remain critically understudied and highlight where future cryo-ET research would be most impactful.

HPF↗

Monte Carlo Simulation of Electron Interactions in an MeV-STEM for Thick Frozen Biological Sample Imaging

A variety of volume electron microscopy techniques have been developed to visualize thick biological samples. However, the resolution is limited by the sliced section thickness (>30–60 nm). To preserve biological samples in a hydrated state, cryo-focused ion beam scanning electron microscopy has been developed, providing nm resolutions. However, this method is time-consuming, requiring 15–20 h to image a 10 μm thick sample with an 8 nm slice thickness. There is a pressing need for a method that allows the rapid and efficient study of thick biological samples while maintaining nanoscale resolution. The remarkable ability of mega-electron-volt (MeV) electrons to penetrate thick biological samples, even exceeding 10 μm in thickness, while maintaining nanoscale resolution, positions MeV-STEM as a suitable microscopy tool for such applications. Our research delves into understanding the interactions between MeV electrons and frozen biological specimens through Monte Carlo simulations. Single elastic scattering, plural elastic scattering, single inelastic scattering, and plural inelastic scattering events have been simulated. The electron trajectories, the beam profile, and the intensity change of electrons in each category have been investigated. Additionally, the effects of the detector collection angle and the focal position of the electron beam were investigated. As electrons penetrated deeper into the specimen, single and plural elastic scattered electrons diminished, and plural inelastic scattered electrons became dominant, and the beam profile became wider. Even after 10 μm of the specimen, 42% of the MeV electrons were collected within 10 mrad. This confirms that MeV-STEM can be employed to study thick biological samples.

electron specimen interaction↗