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Living systems are intrinsically nonequilibrium: They use metabolically derived chemical energy to power their emergent dynamics and self-organization. A crucial driver of these dynamics is the cellular cytoskeleton, a defining example of an active material where the energy injected by molecular motors cascades across length scales, allowing the material to break the constraints of thermodynamic equilibrium and display emergent nonequilibrium dynamics only possible due to the constant influx of energy. Notwithstanding recent experimental advances in the use of local probes to quantify entropy production and the breaking of detailed balance, little is known about the energetics of active materials or how energy propagates from the molecular to emergent length scales. Here, we use a recently developed picowatt calorimeter to experimentally measure the energetics of an active microtubule gel that displays emergent large-scale flows. We find that only approximately one-billionth of the system's total energy consumption contributes to these emergent flows. We develop a chemical kinetics model that quantitatively captures how the system's total thermal dissipation varies with ATP and microtubule concentrations but that breaks down at high motor concentration, signaling an interference between motors. Finally, we estimate how energy losses accumulate across scales. Taken together, these results highlight energetic efficiency as a key consideration for the engineering of active materials and are a powerful step toward developing a nonequilibrium thermodynamics of living systems.
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Citoesqueleto , Microtúbulos , Termodinâmica , Entropia , Modelos QuímicosRESUMO
Barocaloric effectsâsolid-state thermal changes induced by the application and removal of hydrostatic pressureâoffer the potential for energy-efficient heating and cooling without relying on volatile refrigerants. Here, we report that dialkylammonium halidesâorganic salts featuring bilayers of alkyl chains templated through hydrogen bonds to halide anionsâdisplay large, reversible, and tunable barocaloric effects near ambient temperature. The conformational flexibility and soft nature of the weakly confined hydrocarbons give rise to order-disorder phase transitions in the solid state that are associated with substantial entropy changes (>200 J kg-1 K-1) and high sensitivity to pressure (>24 K kbar-1), the combination of which drives strong barocaloric effects at relatively low pressures. Through high-pressure calorimetry, X-ray diffraction, and Raman spectroscopy, we investigate the structural factors that influence pressure-induced phase transitions of select dialkylammonium halides and evaluate the magnitude and reversibility of their barocaloric effects. Furthermore, we characterize the cyclability of thin-film samples under aggressive conditions (heating rate of 3500 K s-1 and over 11,000 cycles) using nanocalorimetry. Taken together, these results establish dialkylammonium halides as a promising class of pressure-responsive thermal materials.
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MicroRNAs (miRNAs) are endogenous and noncoding single-stranded RNA molecules with a length of approximately 18-25 nucleotides, which play an undeniable role in early cancer screening. Therefore, it is very important to develop an ultrasensitive and highly specific method for detecting miRNAs. Here, we present a bottom-up assembly approach for modifying glass microtubes with silica nanowires (SiNWs) and develop a label-free sensing platform for miRNA-21 detection. The three-dimensional (3D) networks formed by SiNWs make them abundant and highly accessible sites for binding with peptide nucleic acid (PNA). As a receptor, PNA has no phosphate groups and exhibits an overall electrically neutral state, resulting in a relatively small repulsion between PNA and RNA, which can improve the hybridization efficiency. The SiNWs-filled glass microtube (SiNWs@GMT) sensor enables ultrasensitive, label-free detection of miRNA-21 with a detection limit as low as 1 aM at a detection range of 1 aM-100 nM. Noteworthy, the sensor can still detect miRNA-21 in the range of 102-108 fM in complex solutions containing 1000-fold homologous interference of miRNAs. The high anti-interference performance of the sensor enables it to specifically recognize target miRNA-21 in the presence of other miRNAs and distinguish 1-, 3-mismatch nucleotide sequences. Significantly, the sensor platform is able to detect miRNA-21 in the lysate of breast cancer cell lines (e.g., MCF-7 cells and MDA-MB-231 cells), indicating that it has good potential in the screening of early breast cancers.
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Vidro , MicroRNAs , Nanofios , Ácidos Nucleicos Peptídicos , Dióxido de Silício , MicroRNAs/análise , Ácidos Nucleicos Peptídicos/química , Dióxido de Silício/química , Humanos , Nanofios/química , Vidro/química , Técnicas Biossensoriais/métodos , Limite de DetecçãoRESUMO
In vivo sensing of the dynamics of ions with high selectivity is essential for gaining molecular insights into numerous physiological and pathological processes. In this work, we report an ion-selective micropipette sensor (ISMS) through the integration of functional crown ether-encapsulated metal-organic frameworks (MOFs) synthesized in situ within the micropipette tip. The ISMS features distinctive sodium ion (Na+) conduction and high selectivity toward Na+ sensing. The selectivity is attributed to the synergistic effects of subnanoconfined space and the specific coordination of 18-crown-6 toward potassium ions (K+), which largely increase the steric hindrance and transport resistance for K+ to pass through the ISMS. Furthermore, the ISMS exhibits high stability and sensitivity, facilitating real-time monitoring of Na+ dynamics in the living rat brain during spreading of the depression events process. In light of the diversity of crown ethers and MOFs, we believe this study paves the way for a nanofluidic platform for in vivo sensing and neuromorphic electrochemical sensing.
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Éteres de Coroa , Estruturas Metalorgânicas , Éteres de Coroa/química , Sódio/química , Íons/química , Potássio/químicaRESUMO
Quantitative phase contrast microscopy (QPCM) can realize high-quality imaging of sub-organelles inside live cells without fluorescence labeling, yet it requires at least three phase-shifted intensity images. Herein, we combine a novel convolutional neural network with QPCM to quantitatively obtain the phase distribution of a sample by only using two phase-shifted intensity images. Furthermore, we upgraded the QPCM setup by using a phase-type spatial light modulator (SLM) to record two phase-shifted intensity images in one shot, allowing for real-time quantitative phase imaging of moving samples or dynamic processes. The proposed technique was demonstrated by imaging the fine structures and fast dynamic behaviors of sub-organelles inside live COS7 cells and 3T3 cells, including mitochondria and lipid droplets, with a lateral spatial resolution of 245â nm and an imaging speed of 250 frames per second (FPS). We imagine that the proposed technique can provide an effective way for the high spatiotemporal resolution, high contrast, and label-free dynamic imaging of living cells.
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Aprendizado Profundo , Imageamento Quantitativo de Fase , Animais , Camundongos , Mitocôndrias , Gotículas LipídicasRESUMO
Staphylococcus aureus (S. aureus) pneumonia has become an increasingly important public health problem. Recent evidence suggests that epigenetic modifications are critical in the host immune defence against pathogen infection. In this study, we found that S. aureus infection induces the expression of histone deacetylase 6 (HDAC6) in a dose-dependent manner. Furthermore, by using a S. aureus pneumonia mouse model, we showed that the HDAC6 inhibitor, tubastatin A, demonstrates a protective effect in S. aureus pneumonia, decreasing the mortality and destruction of lung architecture, reducing the bacterial burden in the lungs and inhibiting inflammatory responses. Mechanistic studies in primary bone marrow-derived macrophages demonstrated that the HDAC6 inhibitors, tubastatin A and tubacin, reduced the intracellular bacterial load by promoting bacterial clearance rather than regulating phagocytosis. Finally, N-acetyl-L- cysteine, a widely used reactive oxygen species (ROS) scavenger, antagonized ROS production and significantly inhibited tubastatin A-induced S. aureus clearance. These findings demonstrate that HDAC6 inhibitors promote the bactericidal activity of macrophages by inducing ROS, an important host factor for S. aureus clearance and production. Our study identified HDAC6 as a suitable epigenetic modification target for preventing S. aureus infection, and tubastatin A as a useful compound in treating S. aureus pneumonia.
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Desacetilase 6 de Histona , Inibidores de Histona Desacetilases , Macrófagos , Espécies Reativas de Oxigênio , Staphylococcus aureus , Animais , Desacetilase 6 de Histona/antagonistas & inibidores , Desacetilase 6 de Histona/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Staphylococcus aureus/efeitos dos fármacos , Camundongos , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Macrófagos/microbiologia , Inibidores de Histona Desacetilases/farmacologia , Ácidos Hidroxâmicos/farmacologia , Pneumonia Estafilocócica/tratamento farmacológico , Pneumonia Estafilocócica/microbiologia , Pneumonia Estafilocócica/metabolismo , Indóis/farmacologia , Camundongos Endogâmicos C57BL , Fagocitose/efeitos dos fármacos , Pulmão/efeitos dos fármacos , Pulmão/microbiologia , Pulmão/metabolismo , Pulmão/patologiaRESUMO
The construction of gating system in artificial channels is a cutting-edge research direction in understanding biological process and application sensing. Here, by mimicking the gating system, we report a device that easily synthesized single-glass micropipettes functionalized by three-dimensional (3D) DNA network, which triggers the gating mechanism for the detection of biomolecules. Based on this strategy, the gating mechanism shows that single-glass micropipette assembled 3D DNA network is in the "OFF" state, and after collapsing in the presence of ATP, they are in the "ON" state, at which point they exhibit asymmetric response times. In the "ON" process of the gating mechanism, the ascorbic acid phosphate (AAP) can be encapsulated by a 3D DNA network and released in the presence of adenosine triphosphate (ATP), which initiates a catalyzed cascade reaction under the influence of alkaline phosphatase (ALP). Ultimately, the detection of ALP can be responded to form the fluorescence signal generated by terephthalic acid that has captured hydroxyl radicals, which has a detection range of 0-250 mU/mL and a limit of detection of 50 mU/mL. This work provides a brand-new way and application direction for research of gating mechanism.
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Trifosfato de Adenosina , Fosfatase Alcalina , DNA , Trifosfato de Adenosina/análise , Trifosfato de Adenosina/química , Trifosfato de Adenosina/metabolismo , Fosfatase Alcalina/metabolismo , Fosfatase Alcalina/química , DNA/química , Vidro/química , Técnicas Biossensoriais/métodos , Limite de Detecção , Ácido Ascórbico/química , Ácido Ascórbico/análogos & derivadosRESUMO
By combining DNA nanotechnology and solid-phase nanopore technology, the aggregation behavior of polymer guided by a single-molecular poly(propylene) (PPO) nucleus in a 3D DNA network has been studied. At low temperature, the PPO chain is evenly dispersed in the rigid 3D DNA network; at higher temperature, the PPO chain self-collapses to a single-molecular nucleus; and upon addition of amphiphilic block copolymers below the critical micelle concentration (CMC), the chains tend to aggregate on the isolated hydrophobic nucleus through intermolecular hydrophobic interactions. The process has been characterized by a rheological test and an electrochemical test. This study not only provides a preliminary understanding of the nucleation and growth process of block copolymers but also offers a theoretical basis for the study of protein self-folding and aggregation in the future. On this basis, utilizing this nucleation and growth event, a novel smart nanopore has been developed for hydrophobicity-dependent molecular transport.
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This paper presents a structured illumination microscopy (SIM) reconstruction algorithm that allows the reconstruction of super-resolved images with 2N + 1 raw intensity images, with N being the number of structured illumination directions used. The intensity images are recorded after using a 2D grating for the projection fringe and a spatial light modulator to select two orthogonal fringe orientations and perform phase shifting. Super-resolution images can be reconstructed from the five intensity images, enhancing the imaging speed and reducing the photobleaching by 17%, compared to conventional two-direction and three-step phase-shifting SIM. We believe the proposed technique will be further developed and widely applied in many fields.
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This study presents a dual-modality microscopic imaging approach that combines quantitative phase microscopy and fluorescence microscopy based on structured illumination (SI) to provide structural and functional information for the same sample. As the first imaging modality, structured illumination digital holographic microscopy (SI-DHM) is implemented along the transmission beam path. SI-DHM acts as a label-free, noninvasive approach and provides high-contrast and quantitative phase images utilizing the refractive index contrast of the inner structures of samples against the background. As the second imaging modality, structured illumination (fluorescence) microscopy (SIM) is constructed along the reflection beam path. SIM utilizes fluorescent labeling and provides super-resolution images for specific functional structures of samples. We first experimentally demonstrated phase imaging of SI-DHM on rice leaves and fluorescence (SIM) imaging on mouse kidney sections. Then, we demonstrated dual-modality imaging of biological samples, using DHM to acquire the overall cell morphology and SIM to obtain specific functional structures. These results prove that the proposed technique is of great importance in biomedical studies, such as providing insight into cell physiology by visualizing and quantifying subcellular structures.
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Holografia , Oryza , Animais , Camundongos , Iluminação , Microscopia de Fluorescência , CorantesRESUMO
Imaging speed and spatial resolution are key factors in optical diffraction tomography (ODT), while they are mutually exclusive in 3D refractive index imaging. This paper presents a multi-harmonic structured illumination-based optical diffraction tomography (MHSI-ODT) to acquire 3D refractive index (RI) maps of transparent samples. MHSI-ODT utilizes a digital micromirror device (DMD) to generate structured illumination containing multiple harmonics. For each structured illumination orientation, four spherical spectral crowns are solved from five phase-shifted holograms, meaning that the acquisition of each spectral crown costs 1.25 raw images. Compared to conventional SI-ODT, which retrieves two spectral crowns from three phase-shifted raw images, MHSI-ODT enhances the imaging speed by 16.7% in 3D RI imaging. Meanwhile, MHSI-ODT exploits both the 1st-order and the 2nd-order harmonics; therefore, it has a better intensity utilization of structured illumination. We demonstrated the performance of MHSI-ODT by rendering the 3D RI distributions of 5 µm polystyrene (PS) microspheres and biological samples.
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As a severe public health concern directly endangering life safety, adolescent suicide has been extensively investigated in variable-centered studies. However, gaps remain in the knowledge of heterogeneous suicide risk patterns and their developmental nature. Additionally, little is known about protective factors associated with suicide risk patterns and changes. This study applied person-centered approaches to explore suicide risk profiles and transitions over time in early Chinese adolescents, along with their protective factors. A total of 1518 junior high school students (49.6% girls, Mage = 13.57, SD = 0.75) participated in two surveys within a 12-month interval. Latent Profile Analysis and Latent Transition Analysis were used to model the profiles and transitions of suicide risk. Three risk profiles were identified at both time points: low risk profile (73.9, 78.3%), medium risk-high threat profile (16.2, 10.2%), and high risk profile (9.9, 10.2%). Low risk profile was stable, while medium risk-high threat and high risk profiles showed great transitions over 12 months. Sense of control, meaning in life, and regulatory emotional self-efficacy served as protective factors against suicide risk profiles and transitions. Findings underscore the importance of comprehensively illustrating suicide risk states from multiple aspects, as well as understanding the fluid nature of transitions between different risk states. Prevention and intervention strategies aimed at enhancing resilience, such as increasing sense of control, perceived meaningfulness, and belief in emotional regulation, may contribute to reducing the risk of suicide among adolescents.
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População do Leste Asiático , Fatores de Proteção , Suicídio , Adolescente , Feminino , Humanos , Masculino , Fatores de Risco , Suicídio/psicologia , Resiliência PsicológicaRESUMO
In this paper, we present large-field, five-step lattice structured illumination microscopy (Lattice SIM). This method utilizes a 2D grating for lattice projection and a spatial light modulator (SLM) for phase shifting. Five phase-shifted intensity images are recorded to reconstruct a super-resolution image, enhancing the imaging speed and reducing the photo-bleaching both by 17%, compared to conventional two-direction and three-shift SIM. Furthermore, lattice SIM has a three-fold spatial bandwidth product (SBP) enhancement compared to SLM/DMD-based SIM, of which the fringe number is limited by the SLM/DMD pixel number. We believe that the proposed technique will be further developed and widely applied in many fields.
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Iluminação , Iluminação/métodos , Microscopia de Fluorescência/métodosRESUMO
Panoramic and long-term observation of nanosized organelle dynamics and interactions with high spatiotemporal resolution still hold great challenge for current imaging platforms. In this study, we propose a live-organelle imaging platform, where a flat-fielding quantitative phase contrast microscope (FF-QPCM) visualizes all the membrane-bound subcellular organelles, and an intermittent fluorescence channel assists in specific organelle identification. FF-QPCM features a high spatiotemporal resolution of 245â nm and 250â Hz and strong immunity against external disturbance. Thus, we could investigate several important dynamic processes of intracellular organelles from direct perspectives, including chromosome duplication in mitosis, mitochondrial fusion and fission, filaments, and vesicles' morphologies in apoptosis. Of note, we have captured, for the first time, a new type of mitochondrial fission (entitled mitochondrial disintegration), the generation and fusion process of vesicle-like organelles, as well as the mitochondrial vacuolization during necrosis. All these results bring us new insights into spatiotemporal dynamics and interactions among organelles, and hence aid us in understanding the real behaviors and functional implications of the organelles in cellular activities.
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Mitocôndrias , Organelas , Microscopia , Microscopia de Contraste de FaseRESUMO
Structured illumination microscopy (SIM) has been widely used in biological research due to its merits of fast imaging speed, minimal invasiveness, super-resolution, and optical sectioning imaging capability. However, the conventional SIM that uses a spatial light modulator (SLM) for fringe projection often has a limited imaging field of view. Herein, we report a large-field SIM technique that combines a 2D grating for fringe pattern projection and an SLM for selecting fringe orientation and performing phase shifting digitally. The proposed SIM technique breaks the bottleneck of fringe number limited by the digital projection devices, while maintaining the advantage of high-speed (digital) phase shifting of conventional SIM. The method avoids the pixilation and dispersion effects of the SLMs. Finally, a 1.8-fold resolution enhancement in a large field of 690 × 517 µm2 under a 20×/NA0.75 objective is experimentally demonstrated. The proposed technique can be widely applied to biology, chemistry, and industry.
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Iluminação , Iluminação/métodos , Microscopia de Fluorescência/métodosRESUMO
The I4-II-IV2-VI7 metal chalcogenide system has become an attractive research system because of its excellent physical and chemical properties. Here, we report the discovery of a new SnII-based quaternary chalcogenide in the I4-II-IV2-VI7 system, Ag4SnGe2S7, with a non-diamond-like structure and crystallizing in the Cc space group. The compound is characterized by isolated pyramid-like [SnS3] units and one-dimensional ∞[SnGe2S8]6- infinite chains with two orientations formed by the corner-sharing connected [SnGe2S8]6- units. It has a band gap of 2.40 eV and is insensitive to air and moisture.
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Quantitative phase microscopy (QPM) is a label-free microscopic technique that exploits the phase of a wave passing through a sample; hence, it has been applied to many fields, including biomedical research and industrial inspection. However, the high spatiotemporal resolution imaging of reflective samples still challenges conventional transmission QPM. In this paper, we propose reflectional quantitative phase-contrast microscopy based on annular epi-illumination of light-emitting diodes. The unscattered wave from the sample is successively phase-retarded by 0, π/2, π, and 3π/2 through a spatial light modulator, and high-resolution phase-contrast images are obtained, revealing the finer structure or three-dimensional tomography of reflective samples. With this system, we have quantitatively obtained the contour of tissue slices and silicon semiconductor wafers. We believe that the proposed system will be very helpful for the high-resolution imaging of industrial devices and biomedical dynamics.
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Iluminação , Silício , Microscopia de Contraste de Fase/métodos , Microscopia/métodos , SemicondutoresRESUMO
It is essential to quantify the physical properties and the dynamics of flowing particles in many fields, especially in microfluidic-related applications. We propose phase image correlation spectroscopy (PICS) as a versatile tool to quantify the concentration, hydro-diameter, and flow velocity of unlabeled particles by correlating the pixels of the phase images taken on flowing particles in a microfluidic device. Compared with conventional image correlation spectroscopy, PICS is minimally invasive, relatively simple, and more efficient, since it utilizes the intrinsic phase of the particles to provide a contrast instead of fluorescent labeling. We demonstrate the feasibility of PICS by measuring flowing polymethylmethacrylate (PMMA) microspheres and yeast in a microfluidic device. We can envisage that PICS will become an essential inspection tool in biomedicine and industry.
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Técnicas Analíticas Microfluídicas , Microfluídica , Polimetil Metacrilato , Dispositivos Lab-On-A-Chip , Microesferas , Análise EspectralRESUMO
This study presents a partially coherent illumination based (PCI-based) SIM apparatus for dual-modality (phase and fluorescent) microscopic imaging. The partially coherent illumination (PCI) is generated by placing a rotating diffuser on a monochromatic laser beam, which suppresses speckle noise in the dual-modality images and endows the apparatus with sound sectioning capability. With this system, label-free quantitative phase and super-resolved/sectioned fluorescent images can be obtained for the same sample. We have demonstrated the superiority of the system in phase imaging of transparent cells with high endogenous contrast and in a quantitative manner. In the meantime, we have also demonstrated fluorescent imaging of fluorescent beads, rat tail crosscut, wheat anther, and hibiscus pollen with super-resolution and optical sectioning. We envisage that the proposed method can be applied to many fields, including but not limited to biomedical, industrial, chemistry fields.
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Function expansion of fiber sensor is highly desired for ultrasensitive optical detection and analysis. Here, we present an approach of multifunctional fiber sensor based on Fano resonances and localized surface plasmon resonance (LSPR) excited via cylindrical vector beam with ability of refractive index (RI) sensing, nano-distance detection, and surface enhanced Raman spectroscopy (SERS). Silver (Ag)-nanocube modified microfiber is theoretically proved to enable to detect RI of the nearby solids and gases based on Fano resonances with a sensitivity of 128.63 nm/refractive index unit (RIU) and 148.21 nm/RIU for solids and gases, respectively. The scattering spectrum of the Ag nanocube has the red-shift response to the varies of the nano-distance between the nanocube and the nearby solid, providing a detection sensitivity up to 1.48 nm (wavelength)/nm (distance). Moreover, this configuration is theoretically verified to have ability to significantly enhance electric field intensity. Radially polarized beam is proved to enhance the electric field intensity as large as 5 times in the side-face configuration compared with linear polarization beam. This fiber-based sensing method is helpful in fields of remote detection, multiple species detection, and cylindrical vector beam-based detection.