Atomic Force Microscopy of the Cell Nucleus

Jiménez‐García, Luis Felipe; Fragoso‐Soriano, Rogelio

doi:10.1006/jsbi.2000.4233

Cited by 18 publications

(14 citation statements)

References 14 publications

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“…In fact, AFM allows us to identify not only large structures such as nuclei but also small ones such as mitochondria and linear chromatin. One similar paper has recently described the clear AFM observation of small structures of rat kidney cells, such as nuclear pores and nucleolus [20]. The results from these two independent studies imply the potential application of AFM to the in situ observation of inner structures in cells.…”

Section: Discussionmentioning

confidence: 75%

“…(ii) AFM is applicable to cells of various sizes, whereas EM cryotomography is currently limited to small cells [2]. AFM allows one to observe structures of various sizes, ranging from large trigeminal ganglion [18] or mouse ES cells (this study) to small nuclei/inner structures [20]. (iii) Serial sections of optional thickness can be visualized using AFM technique.…”

Section: Discussionmentioning

confidence: 99%

“…AFM is also used to compare different material-embedded sections [9] and sections created using various knives [16]. Furthermore, AFM has been employed to image thin-sectioned skeletal muscle [17], semi-thin-sectioned human trigeminal ganglion [18], human oculomotor nerve [19] and epithelial cells of rat kidney [20]. The capacity of AFM to display clear inner structures of cells and/or tissues raises the possibility to visualize nanostructures on serial thin sections of a single cell as well as to perform the 3-D reconstruction of single cells or their inner structures.…”

Section: Introductionmentioning

confidence: 99%

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Atomic force microscopy imaging and 3-D reconstructions of serial thin sections of a single cell and its interior structures

et al. 2005

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The thin sectioning has been widely applied in electron microscopy (EM), and successfully used for an in situ observation of inner ultrastructure of cells. This powerful technique has recently been extended to the research field of atomic force microscopy (AFM). However, there have been no reports describing AFM imaging of serial thin sections and three-dimensional (3-D) reconstruction of cells and their inner structures. In the present study, we used AFM to scan serial thin sections approximately 60nm thick of a mouse embryonic stem (ES) cell, and to observe the in situ inner ultrastructure including cell membrane, cytoplasm, mitochondria, nucleus membrane, and linear chromatin. The high-magnification AFM imaging of single mitochondria clearly demonstrated the outer membrane, inner boundary membrane and cristal membrane of mitochondria in the cellular compartment. Importantly, AFM imaging on six serial thin sections of a single mouse ES cell showed that mitochondria underwent sequential changes in the number, morphology and distribution. These nanoscale images allowed us to perform 3-D surface reconstruction of interested interior structures in cells. Based on the serial in situ images, 3-D models of morphological characteristics, numbers and distributions of interior structures of the single ES cells were validated and reconstructed. Our results suggest that the combined AFM and serial-thin-section technique is useful for the nanoscale imaging and 3-D reconstruction of single cells and their inner structures. This technique may facilitate studies of proliferating and differentiating stages of stem cells or somatic cells at a nanoscale.

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Section: Discussionmentioning

confidence: 75%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Atomic force microscopy imaging and 3-D reconstructions of serial thin sections of a single cell and its interior structures

et al. 2005

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“…Therefore, using sections of material processed for transmission electron microscopy was an approach that produced results. Although initial efforts were made producing images [7], material processed using epoxy resins and semithin sections stained with toluidine blue or even unstained sections, produced images with better Z resolution [8][9][10][11][12][13][14][15][16][17]. Results obtained over the years include samples of plant and animal cell as well as samples from unicellular parasites as Giardia lamblia and Entamoeba histolytica.…”

Section: Introductionmentioning

confidence: 99%

“…Results obtained over the years include samples of plant and animal cell as well as samples from unicellular parasites as Giardia lamblia and Entamoeba histolytica. For example, the plant Lacandonia schismatica, a species with the sex organs inverted, was used as a model to test the technique, producing images similar to those obtained with the light and electron microscopes [8][9][10]13,16]. The cell nucleus and nucleolus were recognized, as well as compact chromatin.…”

Section: Introductionmentioning

confidence: 99%

Visualization of Cell Structure by Atomic Force Microscopy

García¹

2017

MOJAP

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Cell structure has been extensively studied by light and electron microscopy. However, there are relatively few papers on the study of internal cell structure with the atomic force microscope. In this mini review, efforts to visualize and analyze inner cell structure with the atomic force microscope are here presented, using the technique for sample preparation derived from the standard transmission electron microscopy. Semithin sections of epon embedded samples mounted on glass slides are scanned with a microscope working on contact or intermittent modes. The surface of sections revealed internal cell structure. Animal and plant cells show structures as nuclei, nucleoli, chromatin, cell wall, mitochondria. These works opens up the perspective to analyze these organelles and structures at a nanoscale under liquid physiological conditions.

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Chondrocytes interconnecting tracks and cytoplasmic projections observed within the superficial zone of normal human articular cartilage—A transmission electron microscopy, atomic force microscopy, and two‐photon excitation microscopy studies

González¹,

Fragoso‐Soriano²,

Kourí³

2007

Microscopy Res & Technique

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The morphology of the normal human and rat articular cartilage was assessed using transmission electron microscopy (TEM), atomic force microscopy (AFM), and two-photon excitation (2PE) microscopy. Spurr-embedded sections from fixed human cartilage were simultaneously evaluated using TEM and AFM. The presences of tracks among the chondrocytes from the superficial zone of the cartilage were observed. In order to ratify the presence of interconnecting tracks among superficial zone chondrocytes, whole fixed human and rat cartilage, as well as fresh whole rat cartilage, were examined under the 2PE. In all cases, these tracks were observed. In addition, porous matrix, well-defined lacunae, and cytoplasmic projections anchored to the extracellular matrix (ECM) were also detected. We conclude that normal human and rat flattened superficial chondrocytes might be interconnected by tracks running through the ECM. In addition, cytoplasmic projections were observed anchored to the ECM. All these structures may possibly be related to cell/cell and ECM/chondrocytes signaling. Our findings provide new information that possibly will be of relevant importance for a more profound study of normal cartilage physiology and eventually, the pathogenesis of osteoarthritis.

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Atomic Force Microscopy of the Cell Nucleus

Cited by 18 publications

References 14 publications

Atomic force microscopy imaging and 3-D reconstructions of serial thin sections of a single cell and its interior structures

Atomic force microscopy imaging and 3-D reconstructions of serial thin sections of a single cell and its interior structures

Visualization of Cell Structure by Atomic Force Microscopy

Chondrocytes interconnecting tracks and cytoplasmic projections observed within the superficial zone of normal human articular cartilage—A transmission electron microscopy, atomic force microscopy, and two‐photon excitation microscopy studies

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