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Abstract BackgroundIn the past few years, there has been an explosion in single-cell transcriptomics datasets, yet in vivo confirmation of these datasets is hampered in plants due to lack of robust validation methods. Likewise, modeling of plant development is hampered by paucity of spatial gene expression data. RNA fluorescence in situ hybridization (FISH) enables investigation of gene expression in the context of tissue type. Despite development of FISH methods for plants, easy and reliable whole mount FISH protocols have not yet been reported. ResultsWe adapt a 3-day whole mount RNA-FISH method for plant species based on a combination of prior protocols that employs hybridization chain reaction (HCR), which amplifies the probe signal in an antibody-free manner. Our whole mount HCR RNA-FISH method shows expected spatial signals with low background for gene transcripts with known spatial expression patterns in Arabidopsis inflorescences and monocot roots. It allows simultaneous detection of three transcripts in 3D. We also show that HCR RNA-FISH can be combined with endogenous fluorescent protein detection and with our improved immunohistochemistry (IHC) protocol. ConclusionsThe whole mount HCR RNA-FISH and IHC methods allow easy investigation of 3D spatial gene expression patterns in entire plant tissues. 

Abstract The scale of ecological research is getting larger and larger. At such scales, collaboration is indispensable, but there is little consensus on what factors enable collaboration. In the present article, we investigated the temporal and spatial pattern of institutional collaboration within the US Long Term Ecological Research (LTER) Network on the basis of the bibliographic database. Social network analysis and the Monte Carlo method were applied to identify the characteristics of papers published by LTER researchers within a baseline of papers from 158 leading ecological journals. Long-term and long-distance collaboration were more frequent in the LTER Network, and we investigate and discuss the underlying mechanisms. We suggest that the maturing infrastructure and environment for collaboration within the LTER Network could encourage scientists to make large-scale hypotheses and to ask big questions in ecology. 

Abstract This paper presents the design and deployment of the Hydrogen Epoch of Reionization Array (HERA) phase II system. HERA is designed as a staged experiment targeting 21 cm emission measurements of the Epoch of Reionization. First results from the phase I array are published as of early 2022, and deployment of the phase II system is nearing completion. We describe the design of the phase II system and discuss progress on commissioning and future upgrades. As HERA is a designated Square Kilometre Array pathfinder instrument, we also show a number of “case studies” that investigate systematics seen while commissioning the phase II system, which may be of use in the design and operation of future arrays. Common pathologies are likely to manifest in similar ways across instruments, and many of these sources of contamination can be mitigated once the source is identified. 

Abstract We present a framework for identifying and flagging malfunctioning antennas in large radio interferometers. We outline two distinct categories of metrics designed to detect outliers along known failure modes of large arrays: cross‐correlation metrics, based on all antenna pairs, and auto‐correlation metrics, based solely on individual antennas. We define and motivate the statistical framework for all metrics used, and present tailored visualizations that aid us in clearly identifying new and existing systematics. We implement these techniques using data from 105 antennas in the Hydrogen Epoch of Reionization Array (HERA) as a case study. Finally, we provide a detailed algorithm for implementing these metrics as flagging tools on real data sets.