Atomic force microscopy imaging of living cells: a preliminary study of the disruptive effect of the cantilever tip on cell morphology

You, Hong; Lau, Joan M.; Zhang, Shengwen; Yu, Lei

doi:10.1016/s0304-3991(99)00139-4

Cited by 103 publications

(49 citation statements)

References 11 publications

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“…Cells were allowed to attach and spread for at least 16 h. In contrast, other studies examining cell deformation using confocal microscopy investigated nonattached C2C12 myoblasts and chondrocytes, which have a more or less spherical morphology. 6,24 The large diameter of the indenter permitted the compression of the entire cell and not a part of the cell as in the case with other indentation techniques, such as atomic force microscopy 36,42 or cell poking. 26,30 Micropipette aspiration experiments also allow the deformation of the entire cell but are often only applied to suspended cells.…”

Section: Discussionmentioning

confidence: 99%

Anisotropic, Three-Dimensional Deformation of Single Attached Cells Under Compression

Peeters

Bouten

Oomens

et al. 2004

Annals of Biomedical Engineering

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Abstract-Quantifying three-dimensional deformation of cells under mechanical load is relevant when studying cell deformation in relation to cellular functioning. Because most cells are anchorage dependent for normal functioning, it is desired to study cells in their attached configuration. This study reports new threedimensional morphometric measurements of cell deformation during stepwise compression experiments with a recently developed cell loading device. The device allows global, unconfined compression of individual, attached cells under optimal environmental conditions. Three-dimensional images of fluorescently stained myoblasts were recorded with confocal microscopy and analyzed with image restoration and three-dimensional image reconstruction software to quantify cell deformation. In response to compression, cell width, cross-sectional area, and surface area increased significantly with applied strain, whereas cell volume remained constant. Interestingly, the cell and the nucleus deformed perpendicular to the direction of actin filaments present along the long axis of the cell. This strongly suggests that this anisotropic deformation can be attributed to the preferred orientation of actin filaments. A shape factor was introduced to quantify the global shape of attached cells. The increase of this factor during compression reflected the anisotropic deformation of the cell.

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Section: Discussionmentioning

confidence: 99%

Anisotropic, Three-Dimensional Deformation of Single Attached Cells Under Compression

Peeters

Bouten

Oomens

et al. 2004

Annals of Biomedical Engineering

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“…The vertical resolution is mostly determined by the AFM scanner sensitivity, and typically is as high as 0.01nm. The AFM is broadly used to study cell morphology [12][13][14][15][16][17][18][19][20]. However, visualization of the cellular cytoskeleton has been restricted to fibers that are close to the cell surface [18,[21][22][23][24][25][26][27][28][29].…”

Section: Introductionmentioning

confidence: 99%

Visualization of cytoskeletal elements by the atomic force microscope

2005

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We describe a novel application of atomic force microscopy (AFM) to directly visualize cytoskeletal fibers in human foreskin epithelial cells. The nonionic detergent Triton X-100 in a low concentration was used to remove the membrane, soluble proteins, and organelles from the cell. The remaining cytoskeleton can then be directly visualized in either liquid or air-dried ambient conditions. These two types of scanning provide complimentary information. Scanning in liquid visualizes the surface filaments of the cytoskeleton, whereas scanning in air shows both the surface filaments and the total "volume" of the cytoskeletal fibers. The smallest fibers observed were ca. 50 nm in diameter. The lateral resolution of this technique was ca.20 nm, which can be increased to a single nanometer level by choosing sharper AFM tips. Because the AFM is a true three dimensional technique, we are able to quantify the observed cytoskeleton by its density and volume. The types of fibers can be identified by their size, similar to electron microscopy.

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“…The AFM has evolved at an exceptional speed from a laboratory prototype to a commercial instrument (consider, e.g., Veeco Instruments, Woodbury, NY) and many other scanning probe techniques (3) have been developed, which will not be further mentioned here. The AFM has been used to measure forces during stretching of DNA strands (4) and rupture forces of single covalent bonds (5) and can be operated in liquids, which enables its use in biological applications (6)(7)(8). The AFM also can be used to perform surface manipulations such as lithographic fabrication of a transistor (9,10) or AFM-based data storage (11,12).…”

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confidence: 99%

Single-chip mechatronic microsystem for surface imaging and force response studies

Hafizovic

Barrettino

Volden

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

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We report on a stand-alone single-chip (7 ؋ 10 mm) atomic force microscopy unit including a fully integrated array of cantilevers, each of which has an individual actuation, detection, and control unit so that standard atomic force microscopy operations can be performed by means of the chip only without any external controller. The system offers drastically reduced overall size and costs as well as increased scanning speed and can be fabricated with standard complementary metal oxide semiconductor technology with some subsequent micromachining steps to form the cantilevers. Full integration of microelectronic and micromechanical components on the same chip allows for the controlling and monitoring of all system functions. The on-chip circuitry, which includes analog signal amplification and filtering stages with offset compensation, analog-to-digital converters, a powerful digital signal processor, and an on-chip digital interface for data transmission, notably improves the overall system performance. The microsystem characterization evidenced a vertical resolution of <1 nm and a force resolution of <1 nN as shown in the measurement results. The monolithic system represents a paradigm of a mechatronic microsystem that allows for precise and fully controlled mechanical manipulation in the nanoworld.atomic force microscopy ͉ cantilever ͉ complementary metal oxide semiconductor N ew measurement, metrology, and imaging techniques have been pivotal to the rapid development of many branches of science such as materials science, microelectronics, and microbiology, to name a few. The invention of the scanning-tunneling microscope by Binning and Rohrer in 1982 (1) and, in particular, the invention of the atomic force microscope (AFM) in 1986 (2) have established a basis for many findings in various scientific areas over the last years. The AFM has evolved at an exceptional speed from a laboratory prototype to a commercial instrument (consider, e.g., Veeco Instruments, Woodbury, NY) and many other scanning probe techniques (3) have been developed, which will not be further mentioned here. The AFM has been used to measure forces during stretching of DNA strands (4) and rupture forces of single covalent bonds (5) and can be operated in liquids, which enables its use in biological applications (6-8). The AFM also can be used to perform surface manipulations such as lithographic fabrication of a transistor (9, 10) or AFM-based data storage (11,12).Commercially available AFM instruments are rather bulky and have a low throughput because of the serial nature of the involved scanning process and the limited scanning range, which renders the investigation of larger samples rather laborious. The detection of the cantilever deflection is done mostly by means of a laser, which is costly and makes the adjustment and cantilever exchange very time consuming, in particular, when operating in a vacuum environment. To overcome these limitations, AFM probes with integrated detection schemes such as capacitive (13), piezoelectric or piezore...

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Atomic force microscopy imaging of living cells: a preliminary study of the disruptive effect of the cantilever tip on cell morphology

Cited by 103 publications

References 11 publications

Anisotropic, Three-Dimensional Deformation of Single Attached Cells Under Compression

Anisotropic, Three-Dimensional Deformation of Single Attached Cells Under Compression

Visualization of cytoskeletal elements by the atomic force microscope

Single-chip mechatronic microsystem for surface imaging and force response studies

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