Combined biomass network design: A new integrated approach based on ArcGIS.

In light of contemporary energy and environmental objectives, a pivotal transformation of the energy system, encompassing biomass energy, is imperative. A notable challenge in biomass energy facility layout planning is the trade-off between high-efficiency production and the associated investment costs. To harmonize energy efficiency with economy viability, a hybrid layout with the simultaneous construction of centralized and decentralized biomass energy facilities has emerged as a crucial strategic solution. However, the delineation methods for these two layouts lack explicit data support. This study established a population density threshold (PDT) suitable for selecting the distributed layout and employed population density as the criterion for delineating the two layouts. Taking Fuxin City as an example, hybrid layout planning schemes were generated under different PDTs, and a cost and energy benefit analysis framework was developed for these schemes. The results indicated that the scheme with a PDT of 145 person/km2 exhibited the highest energy and economic comprehensive benefits. Compared to a single layout, the planning strategy proposed in this study could achieve nearly the same energy surplus level while saving an investment cost ranging from 2403.9 million CNY to 25,000.23 million CNY. The findings are applicable to other regions with similar conditions, and the analysis framework proposed in this study can be utilized in formulating biomass development strategies for other countries and regions.

On-demand expansion fluorescence and photoacoustic microscopy (ExFLPAM).

Expansion microscopy (ExM) is a promising technology that enables nanoscale imaging on conventional optical microscopes by physically magnifying the specimens. Here, we report the development of a strategy that enables i) on-demand labeling of subcellular organelles in live cells for ExM through transfection of fluorescent proteins that are well-retained during the expansion procedure; and ii) non-fluorescent chromogenic color-development towards efficient bright-field and photoacoustic imaging in both planar and volumetric formats, which is applicable to both cultured cells and biological tissues. Compared to the conventional ExM methods, our strategy provides an expanded toolkit, which we term as expansion fluorescence and photoacoustic microscopy (ExFLPAM), by allowing on-demand fluorescent protein labeling of cultured cells, as well as non-fluorescent absorption contrast-imaging of biological samples.

Advanced 3D imaging and organoid bioprinting for biomedical research and therapeutic applications.

Organoid cultures offer a valuable platform for studying organ-level biology, allowing for a closer mimicry of human physiology compared to traditional two-dimensional cell culture systems or non-primate animal models. While many organoid cultures use cell aggregates or decellularized extracellular matrices as scaffolds, they often lack precise biochemical and biophysical microenvironments. In contrast, three-dimensional (3D) bioprinting allows precise placement of organoids or spheroids, providing enhanced spatial control and facilitating the direct fusion for the formation of large-scale functional tissues in vitro. In addition, 3D bioprinting enables fine tuning of biochemical and biophysical cues to support organoid development and maturation. With advances in the organoid technology and its potential applications across diverse research fields such as cell biology, developmental biology, disease pathology, precision medicine, drug toxicology, and tissue engineering, organoid imaging has become a crucial aspect of physiological and pathological studies. This review highlights the recent advancements in imaging technologies that have significantly contributed to organoid research. Additionally, we discuss various bioprinting techniques, emphasizing their applications in organoid bioprinting. Integrating 3D imaging tools into a bioprinting platform allows real-time visualization while facilitating quality control, optimization, and comprehensive bioprinting assessment. Similarly, combining imaging technologies with organoid bioprinting can provide valuable insights into tissue formation, maturation, functions, and therapeutic responses. This approach not only improves the reproducibility of physiologically relevant tissues but also enhances understanding of complex biological processes. Thus, careful selection of bioprinting modalities, coupled with appropriate imaging techniques, holds the potential to create a versatile platform capable of addressing existing challenges and harnessing opportunities in these rapidly evolving fields.

On the Importance of Low-Frequency Signals in Functional and Molecular Photoacoustic Computed Tomography.

In photoacoustic computed tomography (PACT) with short-pulsed laser excitation, wideband acoustic signals are generated in biological tissues with frequencies related to the effective shapes and sizes of the optically absorbing targets. Low-frequency photoacoustic signal components correspond to slowly varying spatial features and are often omitted during imaging due to the limited detection bandwidth of the ultrasound transducer, or during image reconstruction as undesired background that degrades image contrast. Here we demonstrate that low-frequency photoacoustic signals, in fact, contain functional and molecular information, and can be used to enhance structural visibility, improve quantitative accuracy, and reduce spare-sampling artifacts. We provide an in-depth theoretical analysis of low-frequency signals in PACT, and experimentally evaluate their impact on several representative PACT applications, such as mapping temperature in photothermal treatment, measuring blood oxygenation in a hypoxia challenge, and detecting photoswitchable molecular probes in deep organs. Our results strongly suggest that low-frequency signals are important for functional and molecular PACT.

A method for determining the optimal number and location of biomass energy facilities.

The site selection of biomass energy facilities has always been a key part of energy spatial planning. The site suitability evaluation criteria of the existing studies are not comprehensive. On the other hand, most of the existing studies are to determine the only site, while less research on the multiple-facility planning. The aim of this paper is to identify the most effective number and location for biomass energy facilities. To achieve this objective, the Geographic Information System (GIS) is utilized to perform the following tasks: Generate a site suitability map for potential biomass energy facilities and identify suitable site candidates. The standardization of site suitability evaluation indicators is based on fuzzy logic, and indicator weights are determined based on the Analytic Hierarchy Process (AHP) evaluation of experts' opinions. 2. Develop planning schemes for biomass energy facilities for various number of proposed facilities, and subsequently determine the optimal scheme using multi-objective fuzzy comprehensive evaluation. The weight of each indicator is again determined using the AHP method. Following the analysis, it was found that in the case study of Fuxin City, the plans of 1 and 40 biomass energy facilities can achieve the lowest cost and the highest energy self-sufficiency level. However, both options have potential drawbacks that must be considered. The plan of 30 energy facilities has the highest comprehensive benefits, corresponding to the 30,919.75 yuan of transport cost (3748 yuan lower than the average transport cost) and 75.49% of energy self-sufficiency (67.21% of the average value). This work maximizes the comprehensive positive impacts in economic, environmental and social aspects.

Non-invasive Deep-Brain Imaging with 3D Integrated Photoacoustic Tomography and Ultrasound Localization Microscopy (3D-PAULM).

Photoacoustic computed tomography (PACT) is a proven technology for imaging hemodynamics in deep brain of small animal models. PACT is inherently compatible with ultrasound (US) imaging, providing complementary contrast mechanisms. While PACT can quantify the brain's oxygen saturation of hemoglobin (sO2), US imaging can probe the blood flow based on the Doppler effect. Further, by tracking gas-filled microbubbles, ultrasound localization microscopy (ULM) can map the blood flow velocity with sub-diffraction spatial resolution. In this work, we present a 3D deep-brain imaging system that seamlessly integrates PACT and ULM into a single device, 3D-PAULM. Using a low ultrasound frequency of 4 MHz, 3D-PAULM is capable of imaging the whole-brain hemodynamic functions with intact scalp and skull in a totally non-invasive manner. Using 3D-PAULM, we studied the mouse brain functions with ischemic stroke. Multi-spectral PACT, US B-mode imaging, microbubble-enhanced power Doppler (PD), and ULM were performed on the same mouse brain with intrinsic image co-registration. From the multi-modality measurements, we future quantified blood perfusion, sO2, vessel density, and flow velocity of the mouse brain, showing stroke-induced ischemia, hypoxia, and reduced blood flow. We expect that 3D-PAULM can find broad applications in studying deep brain functions on small animal models.

On the importance of low-frequency signals in functional and molecular photoacoustic computed tomography.

In photoacoustic computed tomography (PACT) with short-pulsed laser excitation, wideband acoustic signals are generated in biological tissues with frequencies related to the effective shapes and sizes of the optically absorbing targets. Low-frequency photoacoustic signal components correspond to slowly varying spatial features and are often omitted during imaging due to the limited detection bandwidth of the ultrasound transducer, or during image reconstruction as undesired background that degrades image contrast. Here we demonstrate that low-frequency photoacoustic signals, in fact, contain functional and molecular information, and can be used to enhance structural visibility, improve quantitative accuracy, and reduce spare-sampling artifacts. We provide an in-depth theoretical analysis of low-frequency signals in PACT, and experimentally evaluate their impact on several representative PACT applications, such as mapping temperature in photothermal treatment, measuring blood oxygenation in a hypoxia challenge, and detecting photoswitchable molecular probes in deep organs. Our results strongly suggest that low-frequency signals are important for functional and molecular PACT.

On-Demand Expansion Fluorescence and Photoacoustic Microscopy (ExFLPAM).

Expansion microscopy (ExM) is a promising technology that enables nanoscale imaging on conventional optical microscopes by physically magnifying the specimens. Here, we report the development of a strategy that enables i) on-demand labeling of subcellular organelles in live cells for ExM through transfection of fluorescent proteins that are well-retained during the expansion procedure; and ii) non-fluorescent chromogenic color-development towards efficient bright-field and photoacoustic imaging in both planar and volumetric formats, which is applicable to both cultured cells and biological tissues. Compared to the conventional ExM methods, our strategy provides an expanded toolkit, which we term as expansion fluorescence and photoacoustic microscopy (ExFLPAM), by allowing on-demand fluorescent protein labeling of cultured cells, as well as non-fluorescent absorption contrast-imaging of biological samples.

Integrated Photoacoustic, Ultrasound, and Angiographic Tomography (PAUSAT) for NonInvasive Whole-Brain Imaging of Ischemic Stroke.

Presented here is an experimental ischemic stroke study using our newly developed noninvasive imaging system that integrates three acoustic-based imaging technologies: photoacoustic, ultrasound, and angiographic tomography (PAUSAT). Combining these three modalities helps acquire multi-spectral photoacoustic tomography (PAT) of the brain blood oxygenation, high-frequency ultrasound imaging of the brain tissue, and acoustic angiography of the cerebral blood perfusion. The multi-modal imaging platform allows the study of cerebral perfusion and oxygenation changes in the whole mouse brain after stroke. Two commonly used ischemic stroke models were evaluated: the permanent middle cerebral artery occlusion (pMCAO) model and the photothrombotic (PT) model. PAUSAT was used to image the same mouse brains before and after a stroke and quantitatively analyze both stroke models. This imaging system was able to clearly show the brain vascular changes after ischemic stroke, including significantly reduced blood perfusion and oxygenation in the stroke infarct region (ipsilateral) compared to the uninjured tissue (contralateral). The results were confirmed by both laser speckle contrast imaging and triphenyltetrazolium chloride (TTC) staining. Furthermore, stroke infarct volume in both stroke models was measured and validated by TTC staining as the ground truth. Through this study, we have demonstrated that PAUSAT can be a powerful tool in noninvasive and longitudinal preclinical studies of ischemic stroke.

Plasmonic nanorod probes' journey inside plant cells for in vivo SERS sensing and multimodal imaging.

Nanoparticle-based platforms are gaining strong interest in plant biology and bioenergy research to monitor and control biological processes in whole plants. However, in vivo monitoring of biomolecules using nanoparticles inside plant cells remains challenging due to the impenetrability of the plant cell wall to nanoparticles beyond the exclusion limits (5-20 nm). To overcome this physical barrier, we have designed unique bimetallic silver-coated gold nanorods (AuNR@Ag) capable of entering plant cells, while conserving key plasmonic properties in the near-infrared (NIR). To demonstrate cellular internalization and tracking of the nanorods inside plant tissue, we used a comprehensive multimodal imaging approach that included transmission electron microscopy (TEM), confocal fluorescence microscopy, two-photon luminescence (TPL), X-ray fluorescence microscopy (XRF), and photoacoustics imaging (PAI). We successfully acquired SERS signals of nanorods in vivo inside plant cells of tobacco leaves. On the same leaf samples, we applied orthogonal imaging methods, TPL and PAI techniques for in vivo imaging of the nanorods. This study first demonstrates the intracellular internalization of AuNR@Ag inside whole plant systems for in vivo SERS analysis in tobacco cells. This work demonstrates the potential of this nanoplatform as a new nanotool for intracellular in vivo biosensing for plant biology.

Multiscale photoacoustic tomography using reversibly switchable thermochromics.

Significance: Based on acoustic detection of optical absorption, photoacoustic tomography (PAT) allows functional and molecular imaging beyond the optical diffusion limit with high spatial resolution. However, multispectral functional and molecular PAT is often limited by decreased spectroscopic accuracy and reduced detection sensitivity in deep tissues, mainly due to wavelength-dependent optical attenuation and inaccurate acoustic inversion. Aim: Previous work has demonstrated that reversible color-shifting can drastically improve the detection sensitivity of PAT by suppressing nonswitching background signals. We aim to develop a new color switching-based PAT method using reversibly switchable thermochromics (ReST). Approach: We developed a family of ReST with excellent water dispersion, biostability, and temperature-controlled color changes by surface modification of commercial thermochromic microcapsules with the hydrophilic polysaccharide alginate. Results: The optical absorbance of the ReST was switched on and off repeatedly by modulating the surrounding temperature, allowing differential photoacoustic detection that effectively suppressed the nonswitching background signal and substantially improved image contrast and detection sensitivity. We demonstrate reversible thermal-switching imaging of ReST in vitro and in vivo using three PAT modes at different length scales. Conclusions: ReST-enabled PAT is a promising technology for high-sensitivity deep tissue imaging of molecular activity in temperature-related biomedical applications, such as cancer thermotherapy.

Three-dimensional non-invasive brain imaging of ischemic stroke by integrated photoacoustic, ultrasound and angiographic tomography (PAUSAT).

We present an ischemic stroke study using our newly-developed PAUSAT system that integrates photoacoustic computed tomography (PACT), high-frequency ultrasound imaging, and acoustic angiographic tomography. PAUSAT is capable of three-dimensional (3D) imaging of the brain morphology, blood perfusion, and blood oxygenation. Using PAUSAT, we studied the hemodynamic changes in the whole mouse brain induced by two common ischemic stroke models: the permanent middle cerebral artery occlusion (pMCAO) model and the photothrombotic (PT) model. We imaged the same mouse brains before and after stroke, and quantitatively compared the two stroke models. We observed clear hemodynamic changes after ischemic stroke, including reduced blood perfusion and oxygenation. Such changes were spatially heterogenous. We also quantified the tissue infarct volume in both stroke models. The PAUSAT measurements were validated by laser speckle imaging and histology. Our results have collectively demonstrated that PAUSAT can be a valuable tool for non-invasive longitudinal studies of neurological diseases at the whole-brain scale.

High-fidelity deep functional photoacoustic tomography enhanced by virtual point sources.

Photoacoustic tomography (PAT), a hybrid imaging modality that acoustically detects the optical absorption contrast, is a promising technology for imaging hemodynamic functions in deep tissues far beyond the traditional optical microscopy. However, the most clinically compatible PAT often suffers from the poor image fidelity, mostly due to the limited detection view of the linear ultrasound transducer array. PAT can be improved by employing highly-absorbing contrast agents such as droplets and nanoparticles, which, however, have low clinical translation potential due to safety concerns and regulatory hurdles imposed by these agents. In this work, we have developed a new methodology that can fundamentally improve PAT's image fidelity without hampering any of its functional capability or clinical translation potential. By using clinically-approved microbubbles as virtual point sources that strongly and isotropically scatter the local pressure waves generated by surrounding hemoglobin, we can overcome the limited-detection-view problem and achieve high-fidelity functional PAT in deep tissues, a technology referred to as virtual-point-source PAT (VPS-PAT). We have thoroughly investigated the working principle of VPS-PAT by numerical simulations and in vitro phantom experiments, clearly showing the signal origin of VPSs and the resultant superior image fidelity over traditional PAT. We have also demonstrated in vivo applications of VPT-PAT for functional small-animal studies with physiological challenges. We expect that VPS-PAT can find broad applications in biomedical research and accelerated translation to clinical impact.

Deep-tissue SWIR imaging using rationally designed small red-shifted near-infrared fluorescent protein.

Applying rational design, we developed 17 kDa cyanobacteriochrome-based near-infrared (NIR-I) fluorescent protein, miRFP718nano. miRFP718nano efficiently binds endogenous biliverdin chromophore and brightly fluoresces in mammalian cells and tissues. miRFP718nano has maximal emission at 718 nm and an emission tail in the short-wave infrared (SWIR) region, allowing deep-penetrating off-peak fluorescence imaging in vivo. The miRFP718nano structure reveals the molecular basis of its red shift. We demonstrate superiority of miRFP718nano-enabled SWIR imaging over NIR-I imaging of microbes in the mouse digestive tract, mammalian cells injected into the mouse mammary gland and NF-kB activity in a mouse model of liver inflammation.

Glassfrogs conceal blood in their liver to maintain transparency.

Transparency in animals is a complex form of camouflage involving mechanisms that reduce light scattering and absorption throughout the organism. In vertebrates, attaining transparency is difficult because their circulatory system is full of red blood cells (RBCs) that strongly attenuate light. Here, we document how glassfrogs overcome this challenge by concealing these cells from view. Using photoacoustic imaging to track RBCs in vivo, we show that resting glassfrogs increase transparency two- to threefold by removing ~89% of their RBCs from circulation and packing them within their liver. Vertebrate transparency thus requires both see-through tissues and active mechanisms that "clear" respiratory pigments from these tissues. Furthermore, glassfrogs' ability to regulate the location, density, and packing of RBCs without clotting offers insight in metabolic, hemodynamic, and blood-clot research.

Intraperitoneal administration for sustained photoacoustic contrast agent imaging.

Photoacoustic (PA) imaging at 1064 nm in the second near-infrared (NIR-II) has attracted recent attention. We recently reported a surfactant-based formulation of a NIR-II dye (BIBDAH) for NIR-II PA contrast. Here, we investigated BIBDAH as a NIR-II PA contrast agent for longitudinal preclinical PA imaging. When administered to mice by the conventional intravenous (I.V.) route, BIBDAH was rapidly cleared from circulation, as indicated by a decrease in NIR-II absorption in sampled plasma. Conversely, when mice were injected with BIBDAH by the intraperitoneal (I.P.) route, peak NIR-II absorption levels in plasma were lower initially, but there was a sustained dye presence that resulted in a more consistent concentration of dye in plasma over 2 days. Increasing the I.P. injection dose and volume resulted in increased NIR-II area under the curve (AUC) in serum. Bimodal PA and ultrasound imaging reflected these results, showing a rapid decrease in PA signal in blood with I.V. administration, but permitting sustained imaging with I.P. administration. These results show that I.P. administration can be considered as an administration route in preclinical animal studies for improved longitudinal observation with more consistent contrast signal intensity.

The effect of biomass raw material collection distance on energy surplus factor.

The collection radius of biomass raw materials is an important factor affecting the volume of raw materials for energy utilization. At present, it is usually studied based on a single biomass combined heat and power (CHP) plant. However, as the heat transfer threshold of biomass CHP plant is limited, it is necessary to consider the optimal collection radius and biomass raw material allocation under the distribution mode of multiple power plants to improve the overall utilization rate of raw materials. Biomass raw material collection distance threshold (BCDT) refers to the maximum road length between the resource point (that allows the transportation of raw materials to the biomass CHP plant) and the biomass CHP plant. Under the mode of multi-power plant planning, the greater the BCDT is, the more destinations there will be for raw materials to be transported to from the same resource point, and the more flexible the transportation plans and allocation of transportation volumes will be. This also means more raw materials can be ultimately used for energy utilization, which leads to higher transportation cost. Therefore, determining the appropriate BCDT plays a key role in the unified planning of biomass raw materials. Based on the limitation of heat transfer threshold, this paper carries out multi-power plant planning with Fuxin City as the research object. Based on such planning, ArcGIS is used to generate biomass raw material planning schemes with different BCDTs. Then the transportation cost and energy surplus factor (ratio of renewable resource potential to energy demand) of each scheme are calculated and compared. The results show that there is a positive correlation between BCDT and the energy surplus factor. With the increase of BCDT, the growth rate of the energy surplus factor gradually becomes slower. The study also allows to set the utilization threshold of biomass energy utilization capacity and obtain the corresponding BCDT. In order to achieve a higher energy surplus factor, it is recommended that 40 km be used as the BCDT when carrying out uniform planning for biomass raw materials. At this time, the utilization of biomass energy utilization capacity is 75%, which can achieve a high degree of energy self-sufficiency and ensure its economic competitiveness.

Optogenetic manipulation and photoacoustic imaging using a near-infrared transgenic mouse model.

Optogenetic manipulation and optical imaging in the near-infrared range allow non-invasive light-control and readout of cellular and organismal processes in deep tissues in vivo. Here, we exploit the advantages of Rhodopseudomonas palustris BphP1 bacterial phytochrome, which incorporates biliverdin chromophore and reversibly photoswitches between the ground (740-800 nm) and activated (620-680 nm) states, to generate a loxP-BphP1 transgenic mouse model. The mouse enables Cre-dependent temporal and spatial targeting of BphP1 expression in vivo. We validate the optogenetic performance of endogenous BphP1, which in the activated state binds its engineered protein partner QPAS1, to trigger gene transcription in primary cells and living mice. We demonstrate photoacoustic tomography of BphP1 expression in different organs, developing embryos, virus-infected tissues and regenerating livers, with the centimeter penetration depth. The transgenic mouse model provides opportunities for both near-infrared optogenetics and photoacoustic imaging in vivo and serves as a source of primary cells and tissues with genomically encoded BphP1.

Three-Dimensional Deep-Tissue Functional and Molecular Imaging by Integrated Photoacoustic, Ultrasound, and Angiographic Tomography (PAUSAT).

Non-invasive small-animal imaging technologies, such as optical imaging, magnetic resonance imaging and x -ray computed tomography, have enabled researchers to study normal biological phenomena or disease progression in their native conditions. However, existing small-animal imaging technologies often lack either the penetration capability for interrogating deep tissues (e.g., optical microscopy), or the functional and molecular sensitivity for tracking specific activities (e.g., magnetic resonance imaging). To achieve functional and molecular imaging in deep tissues, we have developed an integrated photoacoustic, ultrasound and acoustic angiographic tomography (PAUSAT) system by seamlessly combining light and ultrasound. PAUSAT can perform three imaging modes simultaneously with complementary contrast: high-frequency B-mode ultrasound imaging of tissue morphology, microbubble-enabled acoustic angiography of tissue vasculature, and multi-spectral photoacoustic imaging of molecular probes. PAUSAT can provide three-dimensional (3D) multi-contrast images that are co-registered, with high spatial resolutions at large depths. Using PAUSAT, we performed proof-of-concept in vivo experiments on various small animal models: monitoring longitudinal development of placenta and embryo during mouse pregnancy, tracking biodistribution and metabolism of near-infrared organic dye on the whole-body scale, and detecting breast tumor expressing genetically-encoded photoswitchable phytochromes. These results have collectively demonstrated that PAUSAT has broad applicability in biomedical research, providing comprehensive structural, functional, and molecular imaging of small animal models.