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Optical clearing is a versatile approach to improve imaging quality and depth of optical microscopy by reducing scattered light. However, conventional optical clearing methods are restricted in the efficiency-first applications due to unsatisfied time consumption, irreversible tissue deformation, and fluorescence quenching. Here, we developed an ultrafast optical clearing method (FOCM) with simple protocols and common reagents to overcome these limitations. The results show that FOCM can rapidly clarify 300-μm-thick brain slices within 2 min. Besides, the tissue linear expansion can be well controlled by only a 2.12% increase, meanwhile the fluorescence signals of GFP can be preserved up to 86% even after 11 d. By using FOCM, we successfully built the detailed 3D nerve cells model and showed the connection between neuron, astrocyte, and blood vessel. When applied to 3D imaging analysis, we found that the foot shock and morphine stimulation induced distinct c-fos pattern in the paraventricular nucleus of the hypothalamus (PVH). Therefore, FOCM has the potential to be a widely used sample mounting media for biological optical imaging.

Tens of thousands of species are threatened with extinction as a result of human activities. Here we explore how the extinction risks of terrestrial mammals and birds might change in the next 50 years. Future population growth and economic development are forecasted to impose unprecedented levels of extinction risk on many more species worldwide, especially the large mammals of tropical Africa, Asia and South America. Yet these threats are not inevitable. Proactive international efforts to increase crop yields, minimize land clearing and habitat fragmentation, and protect natural lands could increase food security in developing nations and preserve much of Earth's remaining biodiversity.

With high spatio‐temporal resolution geostationary hyperspectral infrared sounder (GeoHIS) observations, monitoring, and predicting rapidly changing weather events are expected to be improved with continuous information of 3D weather cube. However, due to the nature of radiation, clouds prevent an adequate retrieval of atmospheric thermodynamic information under clear‐sky conditions due to large uncertainties in the current radiative transfer model. Removing cloud effects from GeoHIS sub‐footprint is an alternative approach for enhancing clear radiance generation. Such cloud removal can be achieved through the optimal cloud‐clearing (OCC) method, which usually relies on the assumption of atmospheric spatial homogeneity. With high temporal resolution observations from GeoHIS, cloud removal can also be achieved through OCC but using the assumption of temporal homogeneity. This concept was demonstrated using 15‐min Geostationary Interferometric Infrared Sounder (GIIRS) observations. The longwave GIIRS clear radiances under partially cloud cover can be effectively produced with observation errors less than 0.02 ± 0.86 K and 0.04 ± 0.77 K for 11 and 12 μm, respectively. Plain Language Summary With the upcoming high spatio‐temporal resolution geostationary (GEO) hyperspectral infrared (IR) sounder (GeoHIS) observations, the monitoring and prediction of rapidly changing weather events are expected to be improved with continuous information of three‐dimensional weather cube. However, due to the nature of radiation, clouds obscure quantitative applications because of significant uncertainties in the current radiative transfer model. Therefore, enhancing the generation of clear radiance data under partially cloudy conditions is an important alternative approach to improving the quantitative applications of hyperspectral IR data when dealing with clouds. This enhancement of clear radiances has been achieved for hyperspectral IR sounders using the optimal cloud clearing method, which assumes atmospheric spatial homogeneity. Given the unique advantage of high temporal resolution provided by GeoHIS, it is also possible to execute longwave IR clear radiance enhancement through temporal homogeneity assumption for atmospheric temperature. This concept was demonstrated with 15‐min Geostationary Interferometric Infrared Sounder observations in this study and could be used to enhance the quantitative applications of GeoHIS measurements under cloudy sky conditions. Key Points GEO hyperspectral IR sounder longwave clear radiances can be generated under partially cloudy conditions using high temporal information Placing advanced imager and sounder onboard the same GEO platform could enhance the generation of clear radiance data for geostationary hyperspectral infrared sounder (GeoHIS) The concept can be applied to GeoHIS observations if sub‐footprint reference clear data are available from an imager or another source

Organ homeostasis, cellular differentiation, signal relay, and in situ function all depend on the spatial organization of cells in complex tissues. For this reason, comprehensive, high-resolution mapping of cell positioning, phenotypic identity, and functional state in the context of macroscale tissue structure is critical to a deeper understanding of diverse biological processes. Here we report an easy to use method, clearing-enhanced 3D (Cₑ3D), which generates excellent tissue transparency for most organs, preserves cellular morphology and protein fluorescence, and is robustly compatible with antibody-based immunolabeling. This enhanced signal quality and capacity for extensive probe multiplexing permits quantitative analysis of distinct, highly intermixed cell populations in intact Cₑ3D-treated tissues via 3D histo-cytometry. We use this technology to demonstrate large-volume, high-resolution microscopy of diverse cell types in lymphoid and nonlymphoid organs, as well as to perform quantitative analysis of the composition and tissue distribution of multiple cell populations in lymphoid tissues. Combined with histo-cytometry, Cₑ3D provides a comprehensive strategy for volumetric quantitative imaging and analysis that bridges the gap between conventional section imaging and disassociation-based techniques.

Orchids are diverse, occur in a wide range of habitats and dominate threatened species lists, but which orchids are threatened, where and by what? Using the International Union for Conservation of Nature Red List, we assessed the range and diversity of threats to orchids globally including identifying four threat syndromes: (1) terrestrial orchids in forests that are endemic to a country and threatened by illegal collecting; (2) orchids threatened by climate change, pollution, transportation and disturbance/development for tourism, and recreation activities, often in East Asia; (3) epiphytic orchids in Sub-Saharan Africa including Madagascar with diverse threats; and (4) South and Southeast Asia orchids threatened by land clearing for shifting agriculture. Despite limitations in the Red List data, the results highlight how conservation efforts can focus on clusters of co-occurring threats in regions while remaining aware of the trifecta of broad threats from plant collecting, land clearing and climate change.

Histological analysis of biological tissues by mechanical sectioning is significantly time‐consuming and error‐prone due to loss of important information during sample slicing. In the recent years, the development of tissue clearing methods overcame several of these limitations and allowed exploring intact biological specimens by rendering tissues transparent and subsequently imaging them by laser scanning fluorescence microscopy. In this review, we provide a guide for scientists who would like to perform a clearing protocol from scratch without any prior knowledge, with an emphasis on DISCO clearing protocols, which have been widely used not only due to their robustness, but also owing to their relatively straightforward application. We discuss diverse tissue‐clearing options and propose solutions for several possible pitfalls. Moreover, after surveying more than 30 researchers that employ tissue clearing techniques in their laboratories, we compiled the most frequently encountered issues and propose solutions. Overall, this review offers an informative and detailed guide through the growing literature of tissue clearing and can help with finding the easiest way for hands‐on implementation. Graphical Abstract Tissue‐clearing methods have allowed imaging intact biological specimens and understanding biology at a whole‐organism and systems‐level. This Review provides a guide for scientists who would like to setup a clearing protocol, with an emphasis on DISCO methods.

Three-dimensional visualization of tissue structures using optical microscopy facilitates the understanding of biological functions. However, optical microscopy is limited in tissue penetration due to severe light scattering. Recently, a series of tissue-clearing techniques have emerged to allow significant depth-extension for fluorescence imaging. Inspired by these advances, we develop a volumetric chemical imaging technique that couples Raman-tailored tissue-clearing with stimulated Raman scattering (SRS) microscopy. Compared with the standard SRS, the clearing-enhanced SRS achieves greater than 10-times depth increase. Based on the extracted spatial distribution of proteins and lipids, our method reveals intricate 3D organizations of tumor spheroids, mouse brain tissues, and tumor xenografts. We further develop volumetric phasor analysis of multispectral SRS images for chemically specific clustering and segmentation in 3D. Moreover, going beyond the conventional label-free paradigm, we demonstrate metabolic volumetric chemical imaging, which allows us to simultaneously map out metabolic activities of protein and lipid synthesis in glioblastoma. Together, these results support volumetric chemical imaging as a valuable tool for elucidating comprehensive 3D structures, compositions, and functions in diverse biological contexts, complementing the prevailing volumetric fluorescence microscopy.

The clinical use of optical methods for skin imaging is limited by skin strong scattering properties, which reduce image contrast and probing depth. The efficiency of optical methods can be improved by optical clearing (OC). However, for the use of OC agents (OCAs) in a clinical setting, compliance with acceptable non-toxic concentrations is required. OC of human skin, combined with physical and chemical methods to enhance skin permeability to OCAs, was performed to determine the clearing-effectiveness of biocompatible OCAs using line-field confocal optical coherence tomography (LC-OCT) imaging. Nine types of OCAs mixtures were used in association with dermabrasion and sonophoresis for OC protocol on three volunteers hand skin. From 3D images obtained every 5 min for 40 min, the intensity and contrast parameters were extracted to assess their changes during the clearing process and evaluate each OCAs mixture's clearing efficacy. The LC-OCT images average intensity and contrast increased over the entire skin depth with all OCAs. The best image contrast and intensity improvement was observed using the polyethylene glycol, oleic acid, and propylene glycol mixture. Complex OCAs featuring reduced component concentrations that meet drug regulation-established biocompatibility requirements were developed and proved to induce significant skin tissues clearing. By allowing deeper observations and higher contrast, such OCAs in combination with physical and chemical permeation enhancers may improve LC-OCT diagnostic efficacy.