Explore the vast range of titles available.

Acoustic microscopes and acoustic tweezers have great value in the application of microparticle manipulation, biomedical research and non-destructive testing. Ultrahigh frequency (UHF) ultrasonic transducers act as the key component in acoustic microscopes, and acoustic tweezers and acoustic lenses are essential parts of UHF ultrasonic transducers. Therefore, the preparation of acoustic lenses is crucial. Silicon is a suitable material for preparing acoustic lenses because of its high acoustic velocity, low acoustic attenuation and excellent machinability. In previous research, silicon lenses were mainly prepared by etching. However, etching has some drawbacks. The etching of large sizes is complex, time-consuming and expensive. Furthermore, vertical etching is preferred to spherical etching. Thus, a new method of ultra-precision machining was introduced to prepare silicon lenses. In this paper, silicon lenses with an aperture of 892 μm and a depth of 252 μm were prepared. Then, UHF ultrasonic transducers with a center frequency of 157 MHz and a −6-dB bandwidth of 52% were successfully prepared based on silicon lenses. The focal distance of the transducers was 736 μm and the F-number was about 0.82. The transducers had a lateral resolution of 11 μm and could distinguish the 13 μm slots on silicon wafers clearly.

To address the challenge of visualizing internal defects within castings, ultrasonic nondestructive testing technology has been introduced for the detection and characterization of internal defects in castings. Ultrasonic testing is widely utilized for detecting and characterizing internal defects in materials, thanks to its strong penetration ability, wide testing area, and fast scanning speed. However, traditional ultrasonic testing primarily relies on one-dimensional waveforms or two-dimensional images to analyze internal defects in billets, which hinders intuitive characterization of defect quantity, size, spatial distribution, and other relevant information. Consequently, a three-dimensional (3D) layered characterization method of billets internal quality based on scanning acoustic microscope (SAM) is proposed. The method starts with a layered focus scanning of the billet using SAM and pre-processing the obtained sequence of ultrasonic images. Next, the ray casting is employed to reconstruct 3D shape of defects in billets, allowing for characterization of their quality by obtaining characteristic information on defect spatial distributions, quantity, and sizes. To validate the effectiveness of the proposed method, specimens of 42CrMo billets are prepared using five different processes, and the method is employed to evaluate their internal quality. Finally, a comparison between the ultrasonic image and the metallographic image reveals a difference in dimensional accuracy of only 2.94%. The results indicate that the new method enables visualization of internal defect information in billets, serving as a valuable complement to the traditional method of characterizing their internal quality.

Bubbles are ubiquitous in many research applications ranging from ultrasound imaging and drug delivery to the understanding of volcanic eruptions and water circulation in vascular plants. From an acoustic perspective, bubbles are resonant scatterers with remarkable properties, including a large scattering cross-section and strongly sub-wavelength dimensions. While it is known that the resonance properties of bubbles depend on their local environment, it remains challenging to probe this interaction at the single-bubble level due to the difficulty of manipulating a single resonating bubble in a liquid. Here, we confine a cubic bubble inside a cage using 3D printing technology, and we use this bubble as a local probe to perform scanning near-field acoustic microscopy—an acoustic analog of scanning near-field optical microscopy. By probing the acoustic interaction between a single resonating bubble and its local environment, we demonstrate near-field imaging of complex structures with a resolution that is two orders of magnitudes smaller than the wavelength of the acoustic field. As a potential application, our approach paves the way for the development of low-cost acoustic microscopes based on caged bubbles. Scanning a single resonating bubble over structured samples breaks the acoustic diffraction limit. This is made possible by using a bubble enclosed in a 3D-printed cage, that can be robustly manipulated in the near-field of the sample.

The mitochondrial outer membrane contains so-called β-barrel proteins, which allow communication between the cytosol and the mitochondrial interior 1 – 3 . Insertion of β-barrel proteins into the outer membrane is mediated by the multisubunit mitochondrial sorting and assembly machinery (SAM, also known as TOB) 4 – 6 . Here we use cryo-electron microscopy to determine the structures of two different forms of the yeast SAM complex at a resolution of 2.8–3.2 Å. The dimeric complex contains two copies of the β-barrel channel protein Sam50—Sam50a and Sam50b—with partially open lateral gates. The peripheral membrane proteins Sam35 and Sam37 cap the Sam50 channels from the cytosolic side, and are crucial for the structural and functional integrity of the dimeric complex. In the second complex, Sam50b is replaced by the β-barrel protein Mdm10. In cooperation with Sam50a, Sam37 recruits and traps Mdm10 by penetrating the interior of its laterally closed β-barrel from the cytosolic side. The substrate-loaded SAM complex contains one each of Sam50, Sam35 and Sam37, but neither Mdm10 nor a second Sam50, suggesting that Mdm10 and Sam50b function as placeholders for a β-barrel substrate released from Sam50a. Our proposed mechanism for dynamic switching of β-barrel subunits and substrate explains how entire precursor proteins can fold in association with the mitochondrial machinery for β-barrel assembly. Proteins are inserted into the outer mitochondrial membrane by the mitochondrial sorting and assembly machinery, two structural forms of which are presented here, suggesting the mechanism involved.

Cystathionine beta-synthase (CBS) is an essential metabolic enzyme across all domains of life for the production of glutathione, cysteine, and hydrogen sulfide. Appended to the conserved catalytic domain of human CBS is a regulatory domain that modulates activity by S -adenosyl- L -methionine (SAM) and promotes oligomerisation. Here we show using cryo-electron microscopy that full-length human CBS in the basal and SAM-bound activated states polymerises as filaments mediated by a conserved regulatory domain loop. In the basal state, CBS regulatory domains sterically block the catalytic domain active site, resulting in a low-activity filament with three CBS dimers per turn. This steric block is removed when in the activated state, one SAM molecule binds to the regulatory domain, forming a high-activity filament with two CBS dimers per turn. These large conformational changes result in a central filament of SAM-stabilised regulatory domains at the core, decorated with highly flexible catalytic domains. Polymerisation stabilises CBS and reduces thermal denaturation. In PC-3 cells, we observed nutrient-responsive CBS filamentation that disassembles when methionine is depleted and reversed in the presence of SAM. Together our findings extend our understanding of CBS enzyme regulation, and open new avenues for investigating the pathogenic mechanism and therapeutic opportunities for CBS-associated disorders. Cystathionine beta-synthase is a conserved essential enzyme of one-carbon metabolism. Here, the authors show that the enzyme oligomerises to form filaments that undergo conformational and morphological changes in response to its activator S -adenosyl- L -methionine, the global methyl donor.

Three-dimensional photoacoustic imaging (PAM) has emerged as a promising technique for non-invasive label-free visualization and characterization of biological tissues with high spatial resolution and functional contrast. The application of PAM and ultrasound as a microscopy technique of study for Atlantic salmon skin is presented here. A custom ultrasound and photoacoustic experimental setup was used for conducting this experiment with a sample preparation method where the salmon skin is embedded in agarose and lifted from the bottom of the petridish. The results of C-scan, B-scan, and overlayed images of ultrasound and photoacoustic are presented. The results are then analyzed for understanding the pigment map and its relation to salmon behavior to external stimuli. The photoacoustic images are compared with the optical images and analyzed further. A custom colormap and alpha map is designed and the matrices responsible for PAM and ultrasound are inserted together to overlay the ultrasound image and PAM image on top of each other. In this study, we propose an approach that combines scanning acoustic microscopy (SAM) images with PAM images for providing a comprehensive understanding of the salmon skin tissue. Overlaying acoustic and photoacoustic images enabled unique visualization of tissue morphology, with respect to identification of structural features in the context of their pigment distribution.

Lung tissue stiffness is altered with aging. Quantitatively evaluating lung function is difficult using a light microscope (LM) alone. Scanning acoustic microscope (SAM) calculates the speed-of-sound (SOS) using sections to obtain histological images by plotting SOS values on the screen. As SOS is positively correlated with stiffness, SAM has a superior characteristic of simultaneously evaluating tissue stiffness and structure. SOS images of healthy bronchioles, arterioles, and alveoli were compared among young, middle-aged, and old lung sections. Formalin-fixed, paraffin-embedded (FFPE) sections consistently exhibited relatively higher SOS values than fresh-frozen sections, indicating that FFPE became stiffer but retained the relative stiffness reflecting fresh samples. All lung components exhibited gradually declining SOS values with aging and were associated with structural alterations such as loss of smooth muscles, collagen, and elastic fibers. Moreover, reaction to collagenase digestion resulted in decreased SOS values. SOS values of all components were significantly reduced in young and middle-aged groups, whereas no significant reduction was observed in the old group. Protease damage in the absence of regeneration or loss of elastic components was present in old lungs, which exbited dilated bronchioles and alveoli. Aging lungs gradually lose stiffness with decreasing structural components without exposure to specific insults such as inflammation.

Summary This paper focuses on the design of the optimal scanning mode for the family of scanning probe microscopes. Based on different values of the maximum acceleration (deceleration) rate and maximum speed of X‐ and Y‐ axes of the mechanical scanner encountered in practice due to different mechanical design and loads, the design procedure of the optimal fast scanning mode is presented, which is found to be sensitive to the specific parameters of the scanning motion. By utilizing the simultaneous motion of the two axes, the fast raster scanning mode proposed can improve the scanning efficiency by 29% when comparing with the conventional raster (CR) scanning mode, if the scanning speeds of both axes are identical. In addition, the optimal fast mode provided by us has no effects on the image accuracy such as image degradation, image distortion when the efficiency is evaluated. No further difficulties are introduced to the control of the mechanical scanner and the data acquisition process. This optimal scanning mode is useful when the response time of the probe is very fast (such as ultrasonic probe in scanning acoustic microscope (SAM)), and the main limitations are due to the mechanical scanner. By applying different loads for both axes, the experiments with different scanning areas and scanning modes are conducted in a self‐developed SAM. Experimental results coincide with the theoretical analysis and confirm the validation of our proposed optimal fast scanning mode and its superiority over the CR scanning mode. SCANNING 36:185–193, 2014. © 2013 Wiley Periodicals, Inc.

Mitochondrial β-barrel proteins are essential for the transport of metabolites, ions and proteins. The sorting and assembly machinery (SAM) mediates their folding and membrane insertion. We report the cryo-electron microscopy structure of the yeast SAM complex carrying an early eukaryotic β-barrel folding intermediate. The lateral gate of Sam50 is wide open and pairs with the last β-strand (β-signal) of the substrate—the 19-β-stranded Tom40 precursor—to form a hybrid barrel in the membrane plane. The Tom40 barrel grows and curves, guided by an extended bridge with Sam50. Tom40’s first β-segment (β1) penetrates into the nascent barrel, interacting with its inner wall. The Tom40 amino-terminal segment then displaces β1 to promote its pairing with Tom40’s last β-strand to complete barrel formation with the assistance of Sam37’s dynamic α-protrusion. Our study thus reveals a multipoint guidance mechanism for mitochondrial β-barrel folding. The sorting and assembly machinery (SAM) mediates mitochondrial β-barrel protein folding and membrane insertion. A cryo-EM structure of the yeast SAM complex bound to an early eukaryotic β-barrel intermediate reveals a multipoint guidance mechanism.

5,10-methylenetetrahydrofolate reductase (MTHFR) commits folate-derived one-carbon units to generate the methyl-donor s -adenosyl- l -methionine (SAM). Eukaryotic MTHFR appends to the well-conserved catalytic domain (CD) a unique regulatory domain (RD) that confers feedback inhibition by SAM. Here we determine the cryo-electron microscopy structures of human MTHFR bound to SAM and its demethylated product s -adenosyl- l -homocysteine (SAH). In the active state, with the RD bound to a single SAH, the CD is flexible and exposes its active site for catalysis. However, in the inhibited state the RD pocket is remodelled, exposing a second SAM-binding site that was previously occluded. Dual-SAM bound MTHFR demonstrates a substantially rearranged inter-domain linker that reorients the CD, inserts a loop into the active site, positions Tyr404 to bind the cofactor FAD, and blocks substrate access. Our data therefore explain the long-distance regulatory mechanism of MTHFR inhibition, underpinned by the transition between dual-SAM and single-SAH binding in response to cellular methylation status. Here the authors present the cryo-EM structure of active and inhibited human MTHFR, revealing a dynamic inhibitory mechanism dependent on dual SAM binding. The resulting closed conformation features an autoinhibitory element effectively blocking enzymatic activity.