Portal de Periódicos da CAPES

ImageJ for Microscopy

2007; Future Science Ltd; Volume: 43; Issue: sup1 Linguagem: Inglês

10.2144/000112517

1940-9818

Tony Collins,

Radiomics and Machine Learning in Medical Imaging

BioTechniquesVol. 43, No. 1S Imaging FrontiersOpen AccessImageJ for microscopyTony J. CollinsTony J. CollinsMcMaster Biophotonics Facility, McMaster University, Hamilton, ON, CanadaPublished Online:16 May 2018https://doi.org/10.2144/000112517AboutSectionsPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack Citations ShareShare onFacebookTwitterLinkedInRedditEmail IntroductionImageJ will celebrate its tenth anniversary in September of this year. These past 10 years have seen the Java-based open-source software mature into an invaluable laboratory tool. In addition to its impressive functionality, this cutting-edge image-processing tool has an indispensable support community of enthusiasts on the ImageJ mailing list.Wayne Rasband is the core author of ImageJ; after developing the Macintosh-based National Institutes of Health (NIH) Image for 10 years, he made the brave decision of starting afresh with ImageJ using the Java programming language. By shifting to Java, Rasband liberated the software from an individual operating system. To run ImageJ, a given system needs only the operating system-specific Java runtime environment. Java runtime environments (JRE) are freely available, either from Sun or bundled with platform-specific installations of ImageJ (rsb.info.nih.gov/ij). With JRE available for most operating systems, ImageJ is platform-independent, running on Macintosh, Windows, Linux, and even a PDA operating system. The new 64-bit operating systems and their JRE have happily broken the long-held 1.7 Gb memory limit for Java applications. One of the downsides of the Java heritage is an interface that may feel a little unfamiliar. However, a few steps into ImageJ, and this minor inconvenience is forgotten.While Rasband is the author of the core program, an extensive group of additional developers has written and made available a growing arsenal of short add-on programs to provide additional functionality to the core program. These additional files are either written in Java (the plugins) or in ImageJ's macro programming language (macros). Once saved to the ImageJ plugins folder, these functions are loaded on start-up and can be accessed via menu commands like any other core function.400+ PluginsFreely available for individual download, the 400+ plugins contribute to the success of ImageJ, but can also be overwhelming by their sheer magnitude. Where to start? One at a time?The long list of plugins reflects ImageJ's usage throughout a range of fields in science and engineering; it is used in medical imaging, microscopy, the material sciences, not to mention biological light microscopy. A review of the breadth of ImageJ's role in image processing and analysis was published in July 2004 in BioTechniques. This range of applications is reflected in the plugins available. As such, not all are suited for use in microscopy, and some need to be finessed. Needless to say, collecting and maintaining the add-on files that could benefit a given research program would be prohibitively time-consuming and arduous.MBF ImageJThe ImageJ for Microscopy bundle and accompanying manual was developed to manage this wide-ranging array of plugins. Initially collated from the ImageJ home page to help the Laboratory of Molecular Signaling in Babraham Institute (UK), I developed it further at the Wright Cell Imaging Facility (TWRI, Canada); here it was released as WCIF ImageJ. When I recently joined the McMaster Biophotonics Facility (MBF; www.macbiophotonics.ca) at McMaster University, Hamilton, Canada, I was encouraged to maintain this ImageJ for Microscopy bundle.Here the package was resurrected as MBF ImageJ, containing all the plugins that I have found useful. The plugins are organized in submenus, and the bundle is described in an extensively illustrated online manual (which evolved from the original lab instructions). Users of the bundle are encouraged to cite the original authors of the plugins, who have been kind enough to make the results of their work freely available. The online manual provides links to original plugins and authors' pages. In the following discussion, I describe plugins included in the MBF ImageJ bundle (these are freely available on an individual basis elsewhere).The bundle comes in two forms. The first is a one-stop solution for Windows users. This includes a setup file that installs all of the required files. The second version is for non-Windows users. Here, the appropriate version of ImageJ from the ImageJ homepage must be installed followed by a download of the plugins only MBF_ImageJ.zip file (which must be unzipped to a user's ImageJ folder). Each of these approaches will match the installed ImageJ to the version described in the online MBF_ImageJ manual.File FormatsImageJ supports a wide number of standard image file formats, including the recent implementation 48-bit color composite imageJ support. The ability of ImageJ to open a wide variety of proprietary image formats has long been an important feature. Not only is the image data imported, the extra metadata is typically imported as well. This may include useful information, such as exposure settings and laser powers, but also essential settings such as pixel size, acquisition rate, and z-step—all required for proper interpretation of the data.Recently the LOCI group from the University of Wisconsin has developed a bundled suite of plugins that will open over 65 image file formats from the biosciences (see a short list in Table 1 and their web site, www.loci.wisc.edu, for a complete list). This list is continually under development and is continually under development and is frequently updated; the ImageJ community has been providing sample images. An example of the development process reflects the collaborative nature of ImageJ development: when we took possession of our Leica TCS SP5 confocal, it came with yet another proprietary image format. The Leica LIF format is a database file format in which an individual file may contain multiple images (and image series) from a single experiment. Each time the acquisition button is pressed, a new image or image series is saved to the one file. This posed a new challenge for the LOCI group, where a straightforward import plugin would not have been appropriate. A creative solution here was to implement a front end for the LIF file format, prompting the user to select which image to open. A further tweak provides users with the series' name, as well as a helpful thumbnail image of the series. As with previous individual file import plugins, the spatial calibrations are automatically imported, and the full metadata is optionally displayed. This process was greatly facilitated by Leica providing detailed specifications of their new file format—no surprise that the user community widely applauded this action. Manufacturers of acquisition systems are not always so forthcoming with assistance in helping the ImageJ community to generate import plugins for proprietary file formats. The rationale for this is unclear (a restricted image file format is hardly a persuasive reason to choose an acquisition system). It is more easily imagined that, with two equally matched systems, an open image format would be considered a significant advantage.Table 1. Abridged List of Microscopy-Related File Formats Supported by ImageJ Through the LOCI Group's Loci_Tools Plugin SuiteSometimes images need to be imported as TIFF series; ImageJ has a number of sequence import filters and stack manipulation routines to ensure the stack is imported and ordered appropriately for subsequent processing and analysis.A single post-acquisition software solution for a core facility is a huge benefit. Instead of providing routine post-acquisition training for each individual software package, a user trained to process their Bio-Rad PIC images in ImageJ will also be equipped to process their Zeiss AxioVision-acquired images without having to go via viewer software and the dreaded export as tiff step—or the strip metadata step, as I think of it. Bypassing this tedious step when retrieving the original data not only provides immediate access to the image and metadata (facilitating the addition of scale bars for example), it also halves the data storage requirements.Intensity Processing and AnalysisImageJ incorporates a number of useful tools for image processing. These include histogram manipulations and standard image filters (mean, median, etc.)—an excellent background subtraction routine that copes particularly well with uneven background and other user-written plugins for more sophisticated filtering (e.g., Kalman filtering, anisitropic diffusion). Another strength is the large number of automated image segmentation algorithms, again allowing the user to choose the most appropriate. These include Otsu thresholding, mixture modeling, maximum entropy, color-based thresholding, and K-means clustering (this last is particularly good at segmenting color histology images) (see Figure 1).Figure 1. Masson's trichrome-stained heart section.(A) The original image was segmented with three clusters (representing white background, blue collagen, and magenta noncollagen regions). (B) The segmented image is shown with each segment colored to the cluster's centrid value for easy visualization. Scale bar, 100 µm.ImageJ will also perform iterative deconvolution. Bob Dougherty (OptiNav, Bellevue, WA, USA) has written two plugins: one of these generates a theoretical widefield point-spread function (PSF) based on user-provided parameters, and the other plugin uses this PSF to deconvolve a z-series image. It is worth nothing that this plugin requires considerable memory. We have to date only managed to use it with 64-bit operating systems, in which extra memory can be allocated (Figure 2). So, while this plugin is useful for a one-off image, we have not found this to be suitable for routine use.Figure 2. Yeast expressing endoplasmic reticulum (ER)-targeted green fluorescent protein (GFP; green) and stained with Mito Tracker® Orange (red; Invitrogen, Carlsbad, CA, USA).Maximum intensity z-projections of (Ai) the raw data, (Aii) after iterative deconvolution with ImageJ (2 h processing time), and (Aiii) after iterative deconvolution in Volocity (5 min processing time). Scale bar, 2.5 µm. (B) The volume of the mitochondria in the red channel of the Volocity deconvolved data was quantified in ImageJ using the TransfomJ and Object Counter three-dimensional (3-D) plugins. The volume of the mitochondria was calculated as 12.0 µm3 (the volume calculated in Volocity with the same threshold was 12.1 µm3). (Bi) Median section (slice 90) of deconvolved stack overlaid with the object boundaries identified by the ImageJ plugin. (Bii) The axial section along the y-y′ line from BI showing object boundaries in cyan. (Biii) Summed z-projection of the stack of identified object boundaries.The accessibility of the source code has also made ImageJ a favorite for development and implementation of fluorescence resonance energy transfer (FRET) image analysis. There are various plugins available that perform sensitized emission and acceptor photo-bleaching analysis of FRET image sequences. We are also using ImageJ to analyze lifetime images that have been acquired, processed, and exported from our Becker and Hickl TCSPC system.Co-Localization AnalysisAnother key advantage of ImageJ is its abilities in co-localization analysis. Given that no single co-localization quantification technique is appropriate for all circumstances, ImageJ's large suite of co-localization plugins provides options for this additional functionality. Since users are not restricted to a specific approach, the most appropriate co-localization technique from the toolbox can be chosen. Among others, plugins are available to perform qualitative overlays [and convert them from Red-Green-Blue (RGB) to the color-blind friendly Magenta-Green-Blue], generate Pearson's coefficients, generate Mander's coefficients, and perform various randomization co-localization tests [e.g., Fay (1), Costes (2), and van Steensel (3)]. However, as with all analysis tools, they can be misused, and researchers are encouraged to understand these analyses before selecting and using one. The open-source nature of ImageJ also allows the development of novel co-localization routines. Intensity correlation analysis by Li and colleagues was refined only after implementation in ImageJ enabled an improved rate of analysis (4,5).Z-FunctionsIn addition to the standard z-projections and axial sectioning, ImageJ also has some sophisticated three-dimensional (3-D) reconstruction routines. VolumeJ from Michael Abramoff generates ray-traced surface rendering of z-series (Figure 3). This plugin employs a user-defined threshold to generate a surface-rendered image. It can be used to with RGB multichannel or depth-coded images and to generate rotational movies.Figure 3. Three-dimensional (3-D) surface render of depth coded z-series.Confocal z-series of 40 µM thick 4′,6-diamidino-2-phenylindole (DAPI)-stained mouse aorta showing the nuclei in the different smooth muscle layers. Sample provided by M. Kahn (McMaster University). Scale bars, 30 µm.T-FunctionsOne of the quirks of ImageJ is the presumption that a stack's third dimension will be the z-axis. An intensity versus time plot can be performed by the menu command Plot z-axis profile (Figure 4). ImageJ supports a number of time course processing and analysis routines. This includes bleach correction for visualization, normalization to initial intensity, ratio-imaging, and delta-fluorescence routines. The core region of interest (ROI) manager allows multiple regions of interest to be analyzed (and the ROIs saved), greatly facilitating intensity versus time studies, such as calcium imaging. With Rasband's help, we have recently integrated a ratiometric analysis plugin with the ROI manager, again, facilitating analysis, and also providing a plugin template for multi-region processing for other types of analysis.Figure 4. Intensity versus time (intensity vs. T) analysis of a fluo3-loaded HeLa cell treated with 2.5 µM histamine to elicit calcium puffs prior to a global cytoplasmic calcium signal.(A) Median intensity projection of the time-course to illustrate a puffsite (circle) and the axis of the pseudo-linescan plot (x-x′). Scale bar 10, µm. (B) Intensity versus time plot of the puffsite region of interests (ROI; circle in panel A) generated by the Plot z-axis profile menu command. ImageJ assumes the third axis of a stack is z. The raw data was processed for F÷F0 prior to analysis. (C) Intensity versus xt pseudo-linescan image of the line (x-x′) drawn in panel A. Contrast was adjusted to improve visualization of the calcium puffs that precede the global signal. (D) Surface plot of image C using the Interactive three-dimensional (3-D) Surface Plot plugin. (E) Intensity versus xyt. Frames taken from a surface plot movie of the F÷F0 processed image stack generated using the Surface Plot menu command.Particle AnalysisThe integral Particle Analyzer is a powerful multi-region detection and analysis routine. Along with particle separation based on watershed or maxima parameters, ImageJ's core program provides a range of options for users. Plugins add to this functionality with 3-D particle measurements and various particle tracking routines.Macros and PluginsThe internal macro programming language of ImageJ is a simple text-based scripting language that can be used just to automate multiple processing and analysis steps and batch processing, but can also evolve into very sophisticated routines. First attempts at writing macros are easily undertaken using the macro recorder function to record the menu commands a user wants to automate. This initial macro can then be further developed using the macro programming language, which is well documented on the ImageJ web site.The ImageJ web site also hosts many sample macros that can be modified to suit specific needs. There are several generic batch-processing macros that will automate a series of commands on all images in a folder or even all images in subfolders.While macros can become quite sophisticated, the real power of ImageJ is the plugins. As with macros, a user unfamiliar with Java can start by editing the source code of existing plugins.ImageJ ResourcesOnce installed, the core ImageJ program (the IJ.JAR file) can be easily upgraded by simply downloading it from the NIH web site. Keeping an eye on the News page on the ImageJ web site provides a quick summary of what's new with core ImageJ functionality and user-written plugins and macros.The MacBiophotonics ImageJ for Microscopy online manual is a useful resource for new users of ImageJ, covering the typical image processing and analysis steps for microscopists. More complex issues can be addressed via the ImageJ mailing list, an excellent source of information and assistance. The mailing list archives are searchable from the ImageJ web site and usually throw up the answer—many questions have already been addressed. Failing this, a detailed description of the problem to the user group usually results in a quick response with the solution. A recent example of this came when I tried to integrate the ROI manager with a ratio-analysis plugin I had already written. My efforts resulted in a functional, but very slow plugin—taking approximately. I min to analyze a 600-frame stack. A detailed note to the mailing list was met with a quick fix that reduced the analysis to a far more respectable 5 s. This example also addresses one of the perceptions that Java is intrinsically slow. Since Java is highly accessible, functional rather than efficient code can be written by users such as myself with programming-enthusiasm rather than formal training. However, whether a function is executed in 1 or 2 s is probably not relevant except in extreme batch processing of thousands of images.There is an increasing interest in ImageJ at international meetings and workshops. The first ImageJ user and developer conference was held in 2006, and Microscopy and Microanalysis in 2007 will have a session (Symposium A16) dedicated to ImageJ. A number of ImageJ workshops are also cropping up. An ImageJ lab will form an integral part of our Practical Biophotonics introductory workshop that we are hosting in mid-July 2007 and is planned to be a significant part of one of the advanced workshops in the series (www.macbiophotonics.ca/workshop).SummaryImageJ is an essential tool for us that fulfills most of our routine image processing and analysis requirements. The near-comprehensive range of import filters that allow easy access to image and meta-data, a broad suite processing and analysis routine, and enthusiastic support from a friendly mailing list are invaluable for all microscopy labs and facilities—not just those on a budget.AcknowledgmentsI am grateful to Wayne Rasband and the plugin authors who have made their work freely available to the scientific community. MacBiophotonics is supported by the Canadian Foundation for Innovation and the Ontario Innovation Trust.References1. Fay, F.S., K.L. Taneja, S. Shenoy, L. Lifshitz, and R.H. Singer. 1997. Quantitative digital analysis of diffuse and concentrated nuclear distributions of nascent transcripts, SC35 and poly(A). Exp. Cell Res. 231:27–37.Crossref, Medline, CAS, Google Scholar2. Costes, S.V., D. Daelemans, E.H. Cho, Z. Dobbin, G. Pavlakis, and S. Lockett. 2004. Automatic and quantitative measurement of protein-protein colocalization in live cells. Biophys. J. 86:3993–4003.Crossref, Medline, CAS, Google Scholar3. van Steensel, B., E.P. van Binnendijk, C.D. Hornsby, H.T. van der Voort, Z.S. Krozowski, E. R.de Kloet, and R. van Driel. 1996. Partial colocalization of glucocorticoid and mineralocorticoid receptors in discrete compartments in nuclei of rat hippocampus neurons. J. Cell Sci. 109:787–792.Crossref, Medline, CAS, Google Scholar4. Khanna, R., Q. Li, L. Sun, T.J. Collins, and E.F. Stanley. 2006. N type Ca2+ channels and RIM scaffold protein covary at the presynaptic transmitter release face but are components of independent protein complexes. Neuroscience 140:1201–1208.Crossref, Medline, CAS, Google Scholar5. Li, Q., A. Lau, T.J. Morris, L. Guo, C.B. Fordyce, and E.F. Stanley. 2004. A syntaxin 1, Galpha(o), and N-type calcium channel complex at a presynaptic nerve terminal: analysis by quantitative immunocolocalization. J. Neurosci. 24:4070–4081.Crossref, Medline, CAS, Google Scholar6. Dean, P., L. Mascio, D. Ow, D. Sudar, and J. Mullikin. 1990. Proposed standard for image cytometry data files. Cytometry 11:561–569.Crossref, Medline, CAS, Google ScholarFiguresReferencesRelatedDetailsCited ByMachine learning in scanning transmission electron microscopy3 March 2022 | Nature Reviews Methods Primers, Vol. 2, No. 1Numerical learning of deep features from drug-exposed cell images to calculate IC50 without staining22 April 2022 | Scientific Reports, Vol. 12, No. 1Activation of the chemokine receptor 3 pathway leads to a better response to immune checkpoint inhibitors in patients with metastatic urothelial carcinoma13 May 2022 | Cancer Cell International, Vol. 22, No. 1The miR528-D3 Module Regulates Plant Height in Rice by Modulating the Gibberellin and Abscisic Acid Metabolisms20 May 2022 | Rice, Vol. 15, No. 1Application of bacteriophage to develop indicator for Escherichia coli detection and modulation of its biochemical reaction to improve detection ability: A proof-of-concept studyFood Control, Vol. 139Influence of sand transport rate on rolling wear and damage behaviors of wheel/rail in Gobi and desert windblown sand environmentsTribology International, Vol. 172One step nanoencapsulation of corrosion inhibitors for gradual release applicationMaterials Today Chemistry, Vol. 24Reversible and biocompatible AuNP-decorated [Zn2+]:[Insulin] condensed assembly for potential therapeutic applicationsEuropean Journal of Pharmaceutical Sciences, Vol. 173Architectural Terracruda Sculptures of the Silk Roads: New Conservation Insights Through a Diagnostic Approach Based on Non-Destructive X-ray Micro-Computed Tomography17 January 2021 | Studies in Conservation, Vol. 67, No. 4A Magnetosome-Based Platform for Flow Biocatalysis4 May 2022 | ACS Applied Materials & Interfaces, Vol. 14, No. 19AgNP-AC Composite Fibers and its Adsorption and Antibacterial Properties13 May 2022 | Frontiers in Materials, Vol. 9Epithelial Cell-Like Elasticity Modulates Actin-Dependent E-Cadherin Adhesion Organization12 May 2022 | ACS Biomaterials Science & Engineering, Vol. 294Chain Assembly Kinetics from Magnetic Colloidal Spheres29 April 2022 | Langmuir, Vol. 38, No. 18Does Vitamin D Insufficiency Influence Prebiotic Effect on Calcium Absorption and Bone Retention?3 May 2022 | Calcified Tissue International, Vol. 32ATL8, a RING E3 ligase, modulates root growth and phosphate homeostasis in ArabidopsisPlant Physiology and Biochemistry, Vol. 179Antifouling Mortars for Underwater Restoration28 April 2022 | Nanomaterials, Vol. 12, No. 9A deep learning-based method for segmentation and quantitative characterization of microstructures in weathering steel from sequential scanning electron microscope images21 April 2022 | Journal of Iron and Steel Research International, Vol. 29, No. 5RBM20 phosphorylation and its role in nucleocytoplasmic transport and cardiac pathogenesis8 April 2022 | The FASEB Journal, Vol. 36, No. 5Nanoarchitectonics for Ultrathin Gold Films Deposited on Collagen Fabric by High-Power Impulse Magnetron Sputtering10 May 2022 | Nanomaterials, Vol. 12, No. 10DeepRayburst for Automatic Shape Analysis of Tree-Like Structures in Biomedical ImagesIEEE Journal of Biomedical and Health Informatics, Vol. 26, No. 5Removal of Heavy Metals from Industrial Wastewater by Chemical Precipitation: Mechanisms and Sludge Characterization25 March 2021 | Arabian Journal for Science and Engineering, Vol. 47, No. 5Improved Image Analysis for Measuring Gastric Ulcer Index in Animal Models and Clinical Diagnostic Data14 May 2022 | Diagnostics, Vol. 12, No. 5Cholesterol-induced suppression of Kir2 channels is mediated by decoupling at the inter-subunit interfacesiScience, Vol. 25, No. 5Protein Ligand Nanopattern Size Selects for Cellular Adhesion via Hemidesmosomes over Focal Adhesions22 April 2022 | Small Methods, Vol. 62What Is Motion? Recent Advances in the Study of Molecular Movement Patterns of the Peptidoglycan Synthesis MachinesJournal of Bacteriology, Vol. 204, No. 4Association of prostate size with capsule thickness and glandular epithelial cell density: The possible clinical implications on prostate cancer development18 April 2022 | Journal of Clinical Urology, Vol. 12Use of ImageJ Software for Assessment of Mechanical Damage to Starch Granules24 March 2022 | Processes, Vol. 10, No. 4A Portable Nanoprobe for Rapid and Sensitive Detection of SARS-CoV-2 S1 Protein11 April 2022 | Biosensors, Vol. 12, No. 4Diosmetin attenuates oxidative stress-induced damage to lens epithelial cells via the mitogen-activated protein kinase (MAPK) pathway28 April 2022 | Bioengineered, Vol. 13, No. 4RBM20S639G mutation is a high genetic risk factor for premature death through RNA-protein condensatesJournal of Molecular and Cellular Cardiology, Vol. 165Flexible Optogenetic Transducer Device for Remote Neuron Modulation Using Highly Upconversion‐Efficient Dendrite‐Like Gold Inverse Opaline Structure31 March 2022 | Advanced Healthcare Materials, Vol. 55A Novel Lineage of Cile-Like Viruses Discloses the Phylogenetic Continuum Across the Family Kitaviridae28 March 2022 | Frontiers in Microbiology, Vol. 13Powder Metallurgy Route for the Synthesis of Multiprincipal Element Alloys Sputtering Targets27 March 2022 | Advanced Engineering Materials, Vol. 56Petiole anatomy: a contribution to the taxonomy of Acer L. (Sapindaceae) in Iran24 March 2022 | Nordic Journal of Botany, Vol. 14Nanodiamond Integration into Niosomes as an Emerging and Efficient Gene Therapy Nanoplatform for Central Nervous System Diseases15 March 2022 | ACS Applied Materials & Interfaces, Vol. 14, No. 11Improved Preparation of Cu/Zn‐Based Catalysts by Well‐Defined Conditions of Co‐Precipitation and Aging 4 February 2022 | Chemie Ingenieur Technik, Vol. 94, No. 3In search of best automated model: Explaining nanoparticle TEM image segmentationUltramicroscopy, Vol. 233Representativity of morphological measurements and 2-d shape descriptors on mineral admixturesResults in Engineering, Vol. 13Immunohistochemistry as a detection tool for ion channels involved in dental pain signalingThe Saudi Dental Journal, Vol. 34, No. 3The power of weak ion-exchange resins assisted by amelogenin for natural remineralization of dental enamel: an in vitro study11 February 2022 | Odontology, Vol. 28Gender-Associated Oral and Periodontal Health Based on Retrospective Panoramic Radiographic Analysis of Alveolar Bone Loss9 February 2022A food-ingested sublethal concentration of thiamethoxam has harmful effects on the stingless bee Melipona scutellarisChemosphere, Vol. 288Study Effect of nAg Particle Size on the Properties and Antibacterial Characteristics of Polysulfone Membranes25 January 2022 | Nanomaterials, Vol. 12, No. 3Mechanism of Action of the Tungsten Dopant in LiNiO 2 Positive Electrode Materials31 December 2021 | Advanced Energy Materials, Vol. 12, No. 6Comparative genomic analysis of dwarf Vibrio myoviruses defines a conserved gene cluster for successful phage infection8 January 2022 | Archives of Virology, Vol. 167, No. 2A comprehensive landscape of 60S ribosome biogenesis factorsCell Reports, Vol. 38, No. 6Chemogenomics identifies acetyl-coenzyme A synthetase as a target for malaria treatment and preventionCell Chemical Biology, Vol. 29, No. 2Impacts of The Wetland Sedge Carex aquatilis on Microbial Community and Methane Metabolisms2 December 2021 | Plant and Soil, Vol. 471, No. 1-2A conserved arginine in NS5 binds genomic 3′ stem–loop RNA for primer-independent initiation of flavivirus RNA replication10 November 2021 | RNA, Vol. 28, No. 2Density‐dependent patterns of multivariate selection on sperm motility and morphology in a broadcast spawning mussel9 February 2022 | Ecology and Evolution, Vol. 12, No. 2Optical Multilayer Interference Tomography Compatible with Tape-Based Serial SEM for Mesoscale Neuroanatomy15 November 2021 | ACS Photonics, Vol. 9, No. 1Anakinra restores cellular proteostasis by coupling mitochondrial redox balance to autophagy18 January 2022 | Journal of Clinical Investigation, Vol. 132, No. 2Leaf and tuber treatments with PAMPs trigger organ-specific responses in potato22 September 2021 | Canadian Journal of Plant Pathology, Vol. 44, No. 1Effect of introducing manganese as additive on microstructure, hydrogen storage properties and rate limiting step of Ti–Cr alloyInternational Journal of Hydrogen Energy, Vol. 47, No. 1Evaluation of advanced methods and materials for construction of scintillation detector light guidesApplied Radiation and Isotopes, Vol. 179Effect of Multiple Repair Welding on Crack Susceptibility and Mechanical Properties of Inconel 718 Alloy Casting30 August 2021 | Journal of Materials Engineering and Performance, Vol. 31, No. 1Liposome delivery to the brain with rapid short-pulses of focused ultrasound and microbubblesJournal of Controlled Release, Vol. 341Development of a new protocol for a sustainable foliar nutrition in almond orchardsActa Horticulturae, No. 1333Real-time measurement of droplet size and its distribution of an air-induced air-assisted electrostatic nozzleJournal of Electrostatics, Vol. 115Platinum-Nanoparticle-Modified Single-Walled Carbon Nanotube-Laden Paper Electrodes for Electrocatalytic Oxidation of Methanol15 December 2021 | ACS Applied Nano Materials, Vol. 4, No.