High-quality AFM image acquisition of living cells by modified residual encoder-decoder network.

Atomic force microscope enables ultra-precision imaging of living cells. However, atomic force microscope imaging is a complex and time-consuming process. The obtained images of living cells usually have low resolution and are easily influenced by noise leading to unsatisfactory imaging quality, obstructing the research and analysis based on cell images. Herein, an adaptive attention image reconstruction network based on residual encoder-decoder was proposed, through the combination of deep learning technology and atomic force microscope imaging supporting high-quality cell image acquisition. Compared with other learning-based methods, the proposed network showed higher peak signal-to-noise ratio, higher structural similarity and better image reconstruction performances. In addition, the cell images reconstructed by each method were used for cell recognition, and the cell images reconstructed by the proposed network had the highest cell recognition rate. The proposed network has brought insights into the atomic force microscope-based imaging of living cells and cell image reconstruction, which is of great significance in biological and medical research.

Superhydrophobic Antifrosting 7075 Aluminum Alloy Surface with Stable Cassie-Baxter State Fabricated through Direct Laser Interference Lithography and Hydrothermal Treatment.

Frost formation and accumulation can have catastrophic effects on a wide range of industrial activities. Hence, a dual-scale surface with a stable Cassie-Baxter state is developed to mitigate the frosting problem by utilizing direct laser interference lithography assisted with hydrothermal treatment. The high Laplace pressure tolerance under the evaporation stimulus and prolonged Cassie-Baxter state maintenance under the condensation stimulus demonstrate the stable Cassie-Baxter state. The dual-scale surface exhibits a lengthy frost-delaying time of up to 5277 s at -7 °C due to the stable Cassie-Baxter state. The self-removal of frost is achieved by promoting the mobility of frost melts driven by the released interfacial energy. In addition, the dense flocculent frost layer is observed on the single-scale micro surface, whereas the sparse pearl-shaped frost layer with many voids is obtained on the dual-scale surface. This work will aid in understanding the frosting process on various-scale superhydrophobic surfaces and in the design of antifrosting surfaces.

Determining the degree of chromosomal instability in breast cancer cells by atomic force microscopy.

Chromosomal instability (CIN) is a source of genetic variation and is highly linked to the malignance of cancer. Determining the degree of CIN is necessary for understanding the role that it plays in tumor development. There is currently a lack of research on high-resolution characterization of CIN and the relationship between CIN and cell mechanics. Here, a method to determine CIN of breast cancer cells by high resolution imaging with atomic force microscopy (AFM) is explored. The numerical and structural changes of chromosomes in human breast cells (MCF-10A), moderately malignant breast cells (MCF-7) and highly malignant breast cells (MDA-MB-231) were observed and analyzed by AFM. Meanwhile, the nuclei, cytoskeleton and cell mechanics of the three kinds of cells were also investigated. The results showed the differences in CIN between the benign and cancer cells. Also, the degree of structural CIN increased with enhanced malignancy of cancer cells. This was also demonstrated by calculating the probability of micronucleus formation in these three kinds of cells. Meanwhile, we found that the area of the nucleus was related to the number of chromosomes in the nucleus. In addition, reduced or even aggregated actin fibers led to decreased elasticities in MCF-7 and MDA-MB-231 cells. It was found that the rearrangement of actin fibers would affect the nucleus, and then lead to wrong mitosis and CIN. Using AFM to detect chromosomal changes in cells with different malignancy degrees provides a new detection method for the study of cell carcinogenesis with a perspective for targeted therapy of cancer.

Optimizing broadband antireflection with Au micropatterns: a combined FDTD simulation and two-beam LIL approach.

Broadband antireflection (AR) is highly significant in a wide range of optical applications, and using a gold (Au) micropattern presents a viable method for controlling the behavior of light propagation. This study investigates a novel, to the best of our knowledge, methodology to achieve broadband AR properties in Au micropatterns. It employed the three-dimensional finite-difference time-domain (FDTD) method to simulate and optimize the design of micropatterns. In contrast, the fabrication of Au micropatterns was carried out using two-beam laser interference lithography (LIL). The fabricated Au micropatterns were characterized by a scanning electron microscope (SEM) and spectroscope to validate their antireflection and transmission properties and evaluate their performance at various wavelengths. The optimized Au micropatterns had a high transmittance rating of 96.2%. In addition, the device exhibits a broad-spectrum antireflective property, covering wavelengths ranging from 400 to 1100 nm. The simulation data and experimentally derived results show comparable patterns. These structures can potentially be employed in many optical devices, such as solar cells and photodetectors, whereby achieving optimal device performance reduced reflection and enhanced light absorption.

Cell recognition based on atomic force microscopy and modified residual neural network.

Cell recognition methods are in high demand in cell biology and medicine, and the method based on atomic force microscopy (AFM) shows a great value in application. The difference in mechanical properties or morphology of cells has been frequently used to detect whether cells are cancerous, but this detection method cannot be a general means for cancer cell detection, and the traditional artificial feature extraction method also has its limitations. In this work, we proposed an analytic method based on the physical properties of cells and deep learning method for recognizing cell types. The residual neural network used for recognition was modified by multi-scale convolutional fusion, attention mechanism and depthwise separable convolution, so as to optimize feature extraction and reduce operation costs. In the method, the collected cells were imaged by AFM, and the processed images were analyzed by the optimized convolutional neural network. The recognition results of two groups of cells (HL-7702 and SMMC-7721, SGC-7901 and GES-1) by this method show that the recognition rate of dataset with the combination of cell surface morphology, adhesion and Young's modulus is higher, and the recognition rate of the dataset with optimal resolution is higher. Our study indicated that the recognition of physical properties of cells using deep learning technology can serve as a universal and effective method for the automated analysis of cell information.

Large-scale fabrication of an ultrathin broadband absorber using quasi-random dielectric Mie resonators.

Ultrathin broadband absorber maintaining a near-uniform low reflectivity over a broadband wavelength is essential for many optical applications, such as light harvesting and nanoscale imaging. Recently, there has been considerable interest in employing arrays of high-index dielectric Mie resonators on surfaces to trap light and reduce the reflectivity. For such Mie-resonant metasurfaces, however, antireflection properties featuring both a flat low reflectance curve and a wide bandwidth are hard to be satisfied simultaneously, and an efficient large-scale nanofabrication technique rarely exists. Here, we present a high-throughput laser interference induced quasi-random patterning (LIIQP) technique to fabricate quasi-random Mie resonators in large scale. Mie resonators with feature sizes down to sub-100 nm have been fabricated using a 1064 nm laser source. Each Mie resonator concentrates light at its shape-dependent resonant frequency, and all such resonators are arranged quasi-randomly to provide both rich (with broadband Fourier components) and strong (with large intensities) Fourier spectra. Specifically, a near-uniform broadband reflectivity over 400-1100 nm spectrum region has been confined below 3% by fabricating a large-scale ultrathin (around 400 nm) absorber. Our concept and high-throughput fabrication technique allows the rapid production of quasi-random dielectric Mie-resonant metasurfaces in a controllable way, which can be used in various promising applications including thin-film solar cells, display, and imaging.

Cancer Development in Hepatocytes by Long-Term Induction of Hypoxic Hepatocellular Carcinoma Cell (HCC)-Derived Exosomes In Vivo and In Vitro.

Hypoxic tumor cell-derived exosomes play a key role in the occurrence, development, and metastasis of tumors. However, the mechanism of hypoxia-mediated metastasis remains unclear. In this study, hypoxic hepatocellular carcinoma cell (HCC-LM3)-derived exosomes (H-LM3-exos) were used to induce hepatocytes (HL-7702) over a long term (40 passages in 120 days). A nude mouse experiment further verified the effect of H-LM3-exos on tumor growth and metastasis. The process of cancer development in hepatocytes induced by H-LM3-exos was analyzed using both biological and physical techniques, and the results showed that the proliferation and soft agar growth abilities of the transformed cells were enhanced. The concentration of tumor markers secreted by transformed cells was increased, the cytoskeleton was disordered, and the migration ability was enhanced and was accompanied by epithelial-mesenchymal transition (EMT). Transcriptome results showed that differentially expressed genes between transformed cells and hepatocytes were enriched in cancer-related signaling pathways. The degree of cancer development in transformed cells was enhanced by an increase in H-LM3-exos-induced passages. Nude mice treated with different concentrations of H-LM3-exos showed different degrees of tumor growth and liver lesions. The physical properties of the cells were characterized by atomic force microscopy. Compared with the hepatocytes, the height and roughness of the transformed cells were increased, while the adhesion and elastic modulus were decreased. The changes in physical properties of primary tumor cells and hepatocytes in nude mice were consistent with this trend. Our study linking omics with the physical properties of cells provides a new direction for studying the mechanisms of cancer development and metastasis.

Use of Atomic Force Microscopy in UVB-Induced Chromosome Damage Provides Important Bioinformation for Cell Damage Assessment.

The chromosomal structure derived from UVB-stimulated HaCaT cells was detected by atomic force microscopy (AFM) to evaluate the effect of UVB irradiation. The results showed that the higher the UVB irradiation dose, the more the cells that had chromosome aberration. At the same time, different representative types of chromosome structural aberrations were investigated. We also revealed damage to both DNA and cells under the corresponding irradiation doses. It was found that the degree of DNA damage was directly proportional to the irradiation dose. The mechanical properties of cells were also changed after UVB irradiation, suggesting that cells experienced a series of chain reactions from inside to outside after irradiation. The high-resolution imaging of chromosome structures by AFM after UVB irradiation enables us to relate the damage between chromosomes, DNA, and cells caused by UVB irradiation and provides specific information on genetic effects.

Mechanism of action and side effects of colchicine based on biomechanical properties of cells.

Many diseases are related to changes in the biomechanical properties of cells; their study can provide a theoretical basis for drug screening and can explain the internal working of living cells. In this study, the biomechanical properties of nephrocytes (VERO cells), hepatocytes (HL-7702 cells), and hepatoma cells (SMCC-7721 cells) in culture were detected by atomic force microscopy (AFM) to analyse the side effects of colchicine at different concentrations (0.1 µg/mL (A) and 0.2 µg/mL (B)) at the nanoscale for 2, 4 and 6 h. Compared with the corresponding control cells, the damage to the treated cells increased in a dose-dependent manner. Among normal cells, the injury of nephrocytes (VERO cells) was markedly worse than that of hepatocytes (HL-7702 cells) in both colchicine solutions A and B. Based on the analyses of biomechanical properties, the colchicine solution reduced the rate of division and inhibited metastasis of SMCC-7721 cells. By comparing these two concentrations, we found that the anticancer effect of colchicine solution A was greater than that of solution B. Studying the mechanical properties of biological cells can help understand the mechanism of drug action at the molecular level and provide a theoretical basis for preventing the emergence and diagnosis of diseases at the nanoscale.

Repair effect of Centella asiatica (L.) extract on damaged HaCaT cells studied by atomic force microscopy.

People's choice of cosmetics is no longer just 'Follow the trend', but pays more attention to the ingredients of cosmetics, whether the ingredients of cosmetics are beneficial to people's skin health; therefore, more and more skin-healthy ingredients have been discovered and used in cosmetics. In this work, atomic force microscope (AFM) is used to provide physical information about biomolecules and living cells; it brings us a new method of high-precision physical measurement. Centella asiatica (L.) extract has the ability to promote skin wound healing, but its healing effect on damaged HaCaT cells needs to be investigated, which plays a key role in judging the effectiveness of skincare ingredients. The objective of this study was to explore the impact of Centella asiatica (L.) extract on ethanol-damaged human immortalised epidermal HaCaT cells based on AFM. We established a model of cellular damage and evaluated cell viability using the MTT assay. The physical changes of cell height, roughness, adhesion and Young's modulus were measured by AFM. The findings indicated that the Centella asiatica (L.) extract had a good repair effect on injured HaCaT cells, and the optimal concentration was 75 µg/mL.

Comparison of the effects of AgNPs on the morphological and mechanical characteristics of cancerous cells.

Currently, silver nanoparticles (AgNPs) are the most produced nanoparticles in global market and have been widely utilized in the biomedical field. Here, we investigated the morphological and mechanical effects of AgNPs on cancerous cells of A549 cells and SMMC-7721 cells with atomic force microscope (AFM). The influence of AgNPs on the morphological properties and mechanical properties of cancerous cells were characterized utilizing the force-volume (FV) mode and force spectroscopy (FS) mode of AFM measurement. We mainly focus on the comparison of the effects of AgNPs on the two types of cancerous cells based on the fitting results of calculating the Young's moduli utilizing the Sneddon model. The results showed that the morphology changed little, but the mechanical properties of height, roughness, adhesion force and Young's moduli of two cancerous cells varied significantly with the stimulation of different concentrations of AgNPs. This research has provided insights into the classification and characterization of the effects of the various concentrations of AgNPs on the cancerous cells in vitro by utilizing AFM methodologies for disease therapy.

Dynamic morphology imaging of cardiomyocytes based on AFM.

A cardiomyocyte is the basic structural and functional unit of the heart, which is the actual executor of the systolic function. The study of the contraction and relaxation characteristics of cardiomyocyte is of great significance to the physiological behavior and pathology of the heart. How to dynamically express its contraction and relaxation behaviors in 3D has become a challenging issue. Although the video analysis method under the optical microscope can observe the changes in the horizontal direction, it is difficult to describe the changes in the vertical direction. The atomic force microscope (AFM) can accurately express the mechanical and morphological characteristics of the changes in the vertical direction, but it cannot be fully expressed in real time because it is acquired by scanning with a single probe. In order to express the contraction and relaxation characteristics of cardiomyocyte accurately and three dimensionally, a dynamic imaging method in this study is proposed using the periodicity of AFM acquisition and the periodicity of cardiomyocyte contraction. Compared with the optical experiment, it is proven that this method can dynamically represent the contraction and relaxation processes of cardiomyocyte and solve the problem of how to express it in 3D. It brings a new way for the study of physiological characteristics of cardiomyocytes and dynamic imaging by AFM.

MiRNA-520a-3p combined with folic acid conjugated Fe2O3@PDA multifunctional nanoagents for MR imagine and antitumor gene-photothermal therapy.

Osteosarcoma (OS) is a primary malignant bone tumor that occurs mainly in adolescents. Researchers are devoting to develop combination therapy methods in a multifunctional nanoplatform for the treatment of osteosarcoma. The results of previous research have shown that up-regulation of miR-520a-3p could induce anticancer effects in osteosarcoma. In order to improve the effect of gene therapy (GT), we attempted to carry miR-520a-3p in a multifunctional vector for comprehensive therapy. Fe2O3is a type of magnetic resonance imaging (MRI) contrast that is widely used as a drug delivery agent. When coated with polydopamine (PDA), it can also be used as a photothermal therapy (PTT) agent (Fe2O3@ PDA). To deliver nanoagents targeted to a tumor site, folic acid (FA) conjugated with Fe2O3@PDA was manufactured as FA-Fe2O3@PDA. FA was chosen as the target molecule to enhance utilization and reduce toxicity of nanoparticles. However, the therapeutic efficacy of FA-Fe2O3-PDA combined with miR-520a-3p has not yet been studied. In this study, we synthesized FA-Fe2O3@PDA-miRNA and investigated the potential of combining PDA regulated PTT and miR-520a-3p regulated GT to kill osteosarcoma cells. The results indicated that down-regulation of interleukin 6 receptor (IL6R) by miR-520a-3p and the photothermal ability of PDA could induce satisfactory anticancer effects in osteosarcoma, and the curative ratio was better than that used alone PTT or GT. Moreover, as a kind ofT2magnetic contrast, miRNA-Fe2O3@PDA-FA can be used for MRI. These findings indicated that miRNA-Fe2O3@PDA-FA is an effective anti-tumor nanovector for PTT combined with GT.

Study of the Fabrication of Gold Nanoparticle-Graphene-Arrayed Micro/Nanocavities as SERS Substrates Compared to Two Different Angles of Triangular Pyramid Tips.

Surface-enhanced Raman scattering (SERS) has attracted attention because of its enormous potential to detect molecules with low concentrations. The method of fabricating SERS substrates is of great importance for improving the detection resolution. However, SERS substrates with different triangular pyramid tips fabricated by using the tip-based nanoindentation method has not been reported. Here, we prepared arrayed micro/nanocavities on copper-based graphene using the continuous indentation method with a Berkovich tip and a cube-corner tip, which have different face angles. Gold nanoparticles were then sputtered onto the graphene-copper micro/nanocavities to form the Au@GR@Cu micro/nanocavities SERS substrates. The substrates formed using the Berkovich tip and cube-corner tip were labeled B2-B9 and C2-C9, respectively, in which the numbers indicate the machining feed. Rhodamine 6G (R6G) was employed, and the Raman intensities of R6G on the differently arrayed Au@GR@Cu micro/nanocavities were measured. The Raman intensities of R6G were stronger on the pile-ups than on the inverted triangular pyramid cavities. The Raman intensities of R6G were highest on the C2 and B2 structures and lowest on the C9 and B9 structures. The Raman intensities of R6G on the arrayed Au@GR@Cu micro/nanocavities fabricated by the cube-corner tip were stronger than those on the arrayed Au@GR@Cu micro/nanocavities fabricated using the Berkovich tip with the same machining feed. In addition, the electric field intensity and distribution of the B9 and C9 arrayed Au@GR@Cu were simulated using Comsol software. Au@GR@Cu structures fabricated by the cube-corner tip were generated with higher electric field intensities. Furthermore, the relative standard deviations at 1362 cm-1 of R6G were 6.19 and 6.62% on the C2 and C4 surfaces, respectively, showing good homogeneity. The SERS spectra of 10-9 mol/L malachite green solution and 10-6 mol/L carbaryl solution were recognized on the C1, C2, and C4 surfaces on day 1 and after 3 months, respectively. After storage at room temperature for 3 months, the reductions in the Raman intensities were less than 10%, indicating excellent stability. The results showed that the arrayed Au@GR@Cu micro/nanocavities fabricated using the cube-corner tip performed better than those fabricated using the Berkovich tip and exhibited excellent uniformity, availability, and stability, providing great potential for detecting pesticides at low concentrations.

The effects of measurement parameters on the cancerous cell nucleus characterisation by atomic force microscopy in vitro.

Cancer is now responsible for the major leading cause of death worldwide. It is noteworthy that lung cancer has been recognised as the highest incidence (11.6%) and mortality (18.4%) for combined sexes among a variety of cancer diseases. Therefore, it is of great value to investigate the mechanical properties of lung cancerous cells for early diagnosis. This paper focus on the influence of measurement parameters on the measured central Young's moduli of single live A549 cell in vitro based on the force spectroscopy mode of atomic force microscopy (AFM). The effects of the measurement parameters on the measured central Young's moduli were analysed by fitting the force-depth curves utilising the Sneddon model. The results revealed that the Young's moduli of A549 cells increased with the larger indentation force, higher indentation speed, less retraction time, deeper Z length and lower purity percentage of serum. The Young's moduli of cells increased first and then decreased with the increasing dwell time. Hence, this research may have potential significance to provide reference for the standardised detection of a single cancerous cell in vitro using AFM methodologies.

Subsurface phase imaging of tapping-mode atomic force microscopy at phase resonance.

The phase image of tapping-mode atomic force microscopy (TM-AFM) contains energy dissipation, which is related to the sample information on the physical properties such as the sample Young's modulus, adhesion, surface morphology and subsurface morphology. When TM-AFM is used for sample measurement, the frequency near the first resonance peak of probe is usually selected to drive the probe vibration. When the probe vibration is driven by the frequency, the probe has a high amplitude sensitivity, but the phase sensitivity is relatively low. In this paper, the frequency at the probe phase resonance peak was selected for driving the probe vibration to measure the sample, which improved the image resolution. Phase imaging was performed on three uniform photoresist samples with different thicknesses and the same structure. When the scanning parameters were fixed and the probe setpoint value was changed alone, it was found that with the decrease of setpoint value the horizontal resolution of the phase subsurface image was decreased, and the depth sensitivity was increased first and then decreased. The result shows that TM-AFM working at the phase resonance peak can better realise the subsurface imaging of samples at different depths. Phase subsurface imaging at the resonance can be used to quantitatively obtain subsurface phase images of different depths.

Implementation of flexible virtual microchannels based on optically induced dielectrophoresis.

Micro-nano particle manipulation methods in liquid environments have been widely used in the fields such as medicine, biology and material science. Nevertheless, the methods usually rely on pre-prepared physical microfluidic channels. In this work, virtual electrodes based on the optically induced dielectrophoresis (ODEP) method were used as virtual microchannels instead of traditional physical microfluidic channels. Virtual microchannels with different shapes were implemented by the designs of projected light patterns, which made the virtual microchannels have great flexibility and controllability. The theory of ODEP was verified by simulation and analysis of electric field distributions. The relationship between the manipulation force and the alternating current (AC) voltage or the AC frequency exerted on the cells was assessed. The experimental results indicated that the manipulation force was increased with the increase of the AC voltage, and it was reduced with the increase of the AC frequency. Moreover, different virtual microchannels were designed to carry out the transportation, aggregation and sorting of yeast cells and rat basophilic leukemia cells (RBL-2H3 cells) and the survival rate of the cells was evaluated. This work shows that the virtual microchannels can be flexibly realized by ODEP in liquid environments.

Electrical characterization of tumor-derived exosomes by conductive atomic force microscopy.

The physical properties of tumor-derived exosomes have gained much attention because they are helpful to better understand the exosomes in biomedicine. In this study, the conductive atomic force microscopy (C-AFM) was employed to perform the electrical characterizations of exosomes, and it obtained the topography and current images of samples simultaneously. The exosomes were absorbed onto the mica substrates coated with a gold film of 20 nm thick for obtaining the current images of samples by C-AFM in air. The results showed that the single exosomes had the weak conductivity. Furthermore, the currents on exosomes were measured at different bias voltages and pH conditions. It illustrated that the conductivity of exosomes was affected by external factors such as bias voltages and solutions with different pH values. In addition, the electrical responses of low and high metastatic potential cell-derived exosomes were also compared under different voltages and pH conditions. This work is important for better understanding the physical properties of tumor-derived exosomes and promoting the clinical applications of tumor-derived exosomes.

Effect of trypsin concentration on living SMCC-7721 cells studied by atomic force microscopy.

Trypsin is playing an important role in the processes of cancer proliferation, invasion and metastasis which require the precise information of morphology and mechanical properties on the nano-scale for the related research. In this work, living human hepatoma (SMCC-7721) cells were treated with different concentrations of trypsin solution. The morphology and mechanical properties of the cells were measured via atomic force microscope (AFM). Statistical analyses of measurement data indicated that with the increase of trypsin concentration, the average cell height and the surface roughness were both increased, but the cell viability, the cell surface adhesion and the elasticity modulus were decreased significantly. The force required to puncture the cells was also gradually reduced. It indicates that trypsin not only hydrolyses the proteins between the cell and the substrate but also the membrane proteins. The results offer valuable clues for the cancerous process study, pathological analysis and trypsin inhibitor drug development. And this work provides an effective way for overcoming the cell membrane in drug injection for cell-targeted therapy.

High-reliability graphene-wrapped nanoprobes for scanning probe microscopy.

The nanoprobe is a powerful tool in scanning probe microscopy (SPM) that is used to explore various fields of nanoscience. However, the tips can wear out very fast due to the low stability of conventional probes, especially after the measurement of high currents or lateral friction, which results in image distortion and test imprecision. Herein, a novel functional nanoprobe is presented using graphene sheets in a high-quality graphene solution wrapped round a plasma-treated conventional Pt-Ir coated nanoprobe, which shows highly stability and resistance to degradation, leading to a significantly increased lifetime. Furthermore, we show that the graphene-wrapped nanoprobes have the advantages of enhanced electrical conductivity and reduced tip-sample friction, compared with Pt-Ir coated nanoprobes. The simplicity and low cost of this method make it valuable to various functional graphene-wrapped nanoprobes and applications.