scholarly journals Noninvasive Subcellular Imaging Using Atomic Force Acoustic Microscopy (AFAM)

Cells ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 314 ◽  
Author(s):  
Xiaoqing Li ◽  
Ang Lu ◽  
Wenjie Deng ◽  
Li Su ◽  
Jing Wang ◽  
...  
Keyword(s):  
Acoustic Microscopy ◽  
Strong Basis ◽  
Atomic Force ◽  
Cell Structures ◽  
Internal Structures ◽  
Nondestructive Imaging ◽  
Structural Image ◽  
Multiple Samples ◽  
Pseudo Color ◽  
Unique Potential

We report an imaging approach applying the atomic force acoustic microscopy (AFAM), which has unique potential for nondestructive imaging of cell internal structures. To obtain high spatial resolution images, we optimized the significant imaging parameters, including scanning speeds, feedback configurations and acoustic frequencies of an AFAM system, to increase the amplitude of the acoustic signal and to stabilize the morphological signals. We also combined the acoustic amplitude and phase signals, and generated pseudo-color figures for better illustration of subcellular features such as pseudopodia, membranes and nucleus-like. The subcellular structural image atlas can describe nanoscale details of multiple samples and provide clearer images of the subcellular features compared to other conventional techniques. This study builds a strong basis of transmission AFAM for cell imaging, which can help researchers to clarify the cell structures in diverse biological fields and push the understanding of biology evolution to a new stage.

scholarly journals A Fusion Method for Atomic Force Acoustic Microscopy Cell Imaging Based on Local Variance in Non-Subsampled Shearlet Transform Domain

2020 ◽  
Vol 10 (21) ◽  
pp. 7424
Author(s):  
Pengxin Cao ◽  
Xiaoqing Li ◽  
Mingyue Ding
Keyword(s):  
High Frequency ◽  
Low Frequency ◽  
Acoustic Microscopy ◽  
Source Image ◽  
Fusion Method ◽  
Cell Region ◽  
Shearlet Transform ◽  
Local Variance ◽  
Atomic Force ◽  
Internal Structures

Atomic force acoustic microscopy (AFAM) is a measurement method that uses the probe and acoustic wave to image the surface and internal structures of different materials. For cellular material, the morphology and phase images of AFAM reflect the outer surface and internal structures of the cell, respectively. This paper proposes an AFAM cell image fusion method in the Non-Subsampled Shearlet Transform (NSST) domain, based on local variance. First, NSST is used to decompose the source images into low-frequency and high-frequency sub-bands. Then, the low-frequency sub-band is fused by the weight of local variance, while a contrast limited adaptive histogram equalization is used to improve the source image contrast to better express the details in the fused image. The high-frequency sub-bands are fused using the maximum rule. Since the AFAM image background contains a lot of noise, and improved segmentation algorithm based on the Otsu algorithm is proposed to segment the cell region, and the image quality metrics based on the segmented region will make the evaluation more accurate. Experiments with different groups of AFAM cell images demonstrated that the proposed method can clearly show the internal structures and the contours of the cells, compared with traditional methods.

scholarly journals An Atomic Force Acoustic Microscopy Image Fusion Method Based on Grayscale Inversion and Selection of Best-Fit Intensity

2020 ◽  
Vol 10 (23) ◽  
pp. 8645
Author(s):  
Zhaozheng Chen ◽  
Xiaoqing Li ◽  
Mingyue Ding
Keyword(s):  
Mutual Information ◽  
Visual Information ◽  
Acoustic Microscopy ◽  
Atomic Force ◽  
Acoustic Images ◽  
Internal Structures ◽  
Microscopy Image ◽  
Information Fidelity ◽  
Best Fit ◽  
Selection Of

Atomic force acoustic microscopy (AFAM) can provide surface morphology and internal structures of the samples simultaneously, with broad potential in non-destructive imaging of cells. As the output of AFAM, morphology and acoustic images reflect different features of the cells, respectively. However, there are few studies about the fusion of these images. In this paper, a novel method is proposed to fuse these two types of images based on grayscale inversion and selection of best-fit intensity. First, grayscale inversion is used to transform the morphology image into a series of inverted images with different average intensities. Then, the max rule is applied to fuse those inverted images and acoustic images, and a group of pre-fused images is obtained. Finally, a selector is employed to extract and export the expected image with the best-fit intensity among those pre-fused images. The expected image can preserve both the acoustic details of the cells and the background’s gradient information well, which benefits the analysis of the cell’s subcellular structure. The experiments’ results demonstrated that our method could provide the clearest boundaries between the cells and background, and preserve most details from the morphology and acoustic images according to quantitative comparisons, including standard deviation, mutual information, Xydeas and Petrovic metric, feature mutual information, and visual information fidelity fusion.

Atomic force acoustic microscopy characterization of carbon-based materials

2007 ◽  
Author(s):  
D. Passeri ◽  
A. Bettucci ◽  
M. Germano ◽  
A. Biagioni ◽  
M. Rossi ◽  
...  
Keyword(s):  
Acoustic Microscopy ◽  
Atomic Force ◽  
Microscopy Characterization ◽  
Carbon Based

scholarly journals Measurement of the Indentation Modulus and the Local Internal Friction in Amorphous SiO2 Using Atomic Force Acoustic Microscopy

2016 ◽  
Vol 61 (1) ◽  
pp. 9-12
Author(s):  
B. Zhang ◽  
H. Wagner ◽  
M. Büchsenschütz-Göbeler ◽  
Y. Luo ◽  
S. Küchemann ◽  
...  
Keyword(s):  
Internal Friction ◽  
Amorphous Materials ◽  
Contact Stiffness ◽  
Ultrasonic Transducer ◽  
Amorphous Material ◽  
Acoustic Microscopy ◽  
Damping Factor ◽  
Indentation Modulus ◽  
Amorphous Sio2 ◽  
Atomic Force

Abstract For the past two decades, atomic force acoustic microscopy (AFAM), an advanced scanning probe microscopy technique, has played a promising role in materials characterization with a good lateral resolution at micro/nano dimensions. AFAM is based on inducing out-of-plane vibrations in the specimen, which are generated by an ultrasonic transducer. The vibrations are sensed by the AFM cantilever when its tip is in contact with the material under test. From the cantilver’s contactresonance spectra, one determines the real and the imaginary part of the contact stiffness k*, and then from these two quantities the local indentation modulus M' and the local damping factor Qloc-1 can be obtained with a spatial resolution of less than 10 nm. Here, we present measured data of M' and of Qloc-1 for the insulating amorphous material, a-SiO2. The amorphous SiO2 layer was prepared on a crystalline Si wafer by means of thermal oxidation. There is a spatial distribution of the indentation modulus M' and of the internal friction Qloc-1. This is a consequence of the potential energy landscape for amorphous materials.

scholarly journals Movements of Mycoplasma mobile gliding machinery detected by high-speed atomic force microscopy

2021 ◽  
Author(s):  
Kohei Kobayashi ◽  
Noriyuki Kodera ◽  
Taishi Kasai ◽  
Yuhei O Tahara ◽  
Takuma Toyonaga ◽  
...  
Keyword(s):  
Atomic Force Microscopy ◽  
Internal Structure ◽  
Sodium Azide ◽  
High Speed ◽  
Atp Hydrolysis ◽  
Solid Surfaces ◽  
Force Microscopy ◽  
Special Mechanism ◽  
Atomic Force ◽  
Internal Structures

ABSTRACTMycoplasma mobile, a parasitic bacterium, glides on solid surfaces, such as animal cells and glass by a special mechanism. This process is driven by the force generated through ATP hydrolysis on an internal structure. However, the spatial and temporal behaviors of the internal structures in living cells are unclear. In this study, we detected the movements of the internal structure by scanning cells immobilized on a glass substrate using high-speed atomic force microscopy (HS-AFM). By scanning the surface of a cell, we succeeded in visualizing particles, 2 nm in hight and aligned mostly along the cell axis with a pitch of 31.5 nm, consistent with previously reported features based on electron microscopy. Movements of individual particles were then analyzed by HS-AFM. In the presence of sodium azide, the average speed of particle movements was reduced, suggesting that movement is linked to ATP hydrolysis. Partial inhibition of the reaction by sodium azide enabled us to analyze particle behavior in detail, showing that the particles move 9 nm right, relative to the gliding direction, and 2 nm into the cell interior in 330 ms, then return to their original position, based on ATP hydrolysis.IMPORTANCEThe Mycoplasma genus contains bacteria generally parasitic to animals and plants. Some Mycoplasma species form a protrusion at a pole, bind to solid surfaces, and glide by a special mechanism linked to their infection and survival. The special machinery for gliding can be divided into surface and internal structures that have evolved from rotary motors represented by ATP synthases. This study succeeded in visualizing the real-time movements of the internal structure by scanning from the outside of the cell using an innovative high-speed atomic force microscope, and then analyzing their behaviors.

Mechanical characterization of nanoporous materials by use of atomic force acoustic microscopy methods

2013 ◽  
Vol 24 (35) ◽  
pp. 355703 ◽  
Author(s):  
M Kopycinska-Müller ◽  
K-B Yeap ◽  
S Mahajan ◽  
B Köhler ◽  
N Kuzeyeva ◽  
...  
Keyword(s):  
Mechanical Characterization ◽  
Nanoporous Materials ◽  
Acoustic Microscopy ◽  
Atomic Force

Quantitative Contact Spectroscopy by Atomic-Force Acoustic Microscopy

2006 ◽  
pp. 179-186
Author(s):  
U. Rabe ◽  
E. Kester ◽  
V. Scherer ◽  
W. Arnold
Keyword(s):  
Acoustic Microscopy ◽  
Atomic Force

scholarly journals Atomic force acoustic microscopy reveals the influence of substrate stiffness and topography on cell behavior

2019 ◽  
Vol 10 ◽  
pp. 2329-2337 ◽  
Author(s):  
Yan Liu ◽  
Li Li ◽  
Xing Chen ◽  
Ying Wang ◽  
Meng-Nan Liu ◽  
...  
Keyword(s):  
Acoustic Microscopy ◽  
Substrate Stiffness ◽  
Cell Behavior ◽  
Cell Alignment ◽  
Nondestructive Technique ◽  
Elastic Deformations ◽  
L929 Cells ◽  
Atomic Force ◽  
Cell Substrate ◽  
Influence Of Substrate

The stiffness and the topography of the substrate at the cell–substrate interface are two key properties influencing cell behavior. In this paper, atomic force acoustic microscopy (AFAM) is used to investigate the influence of substrate stiffness and substrate topography on the responses of L929 fibroblasts. This combined nondestructive technique is able to characterize materials at high lateral resolution. To produce substrates of tunable stiffness and topography, we imprint nanostripe patterns on undeveloped and developed SU-8 photoresist films using electron-beam lithography (EBL). Elastic deformations of the substrate surfaces and the cells are revealed by AFAM. Our results show that AFAM is capable of imaging surface elastic deformations. By immunofluorescence experiments, we find that the L929 cells significantly elongate on the patterned stiffness substrate, whereas the elasticity of the pattern has only little effect on the spreading of the L929 cells. The influence of the topography pattern on the cell alignment and morphology is even more pronounced leading to an arrangement of the cells along the nanostripe pattern. Our method is useful for the quantitative characterization of cell–substrate interactions and provides guidance for the tissue regeneration therapy in biomedicine.

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