No correlation of focal contacts and close adhesion by comparing GFP-vinculin and fluorescence interference of DiI

Iwanaga, Yoriko; Braun, Dieter; Fromherz, Peter

doi:10.1007/s002490000119

Cited by 65 publications

(58 citation statements)

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“…Our results for the average cleft distance values (table 2) are in good accordance with the previously published distances of mammalian cells on microstructured and protein-coated silicon dioxide surfaces, which were obtained by FLIC microscopy (Braun & Fromherz 1998;Iwanaga et al 2001;Lambacher & Fromherz 2002). The cleft distance of HEK293 cells on different proteins was previously studied by FLIC microscopy: 55G10 nm for collagen (Straub et al 2001), and 70G10 and 75G1 nm, respectively, for FIB (Brittinger & Fromherz 2005;Gleixner & Fromherz 2006).…”

Section: Discussionsupporting

confidence: 91%

Transmission electron microscopy study of the cell–sensor interface

Wrobel

Höller

Ingebrandt

et al. 2007

J. R. Soc. Interface.

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An emerging number of micro-and nanoelectronics-based biosensors have been developed for non-invasive recordings of physiological cellular activity. The interface between the biological system and the electronic devices strongly influences the signal transfer between these systems. Little is known about the nanoscopic structure of the cell-sensor interface that is essential for a detailed interpretation of the recordings. Therefore, we analysed the interface between the sensor surface and attached cells using transmission electron microscopy (TEM). The maximum possible resolution of our TEM study, however, was restricted by the quality of the interface preparation. Therefore, we complemented our studies with imaging ellipsometry.We cultured HEK293 cells on substrates, which had been precoated with different types of proteins. We found that contact geometry between attached cell membrane and substrate was dependent on the type of protein coating used. In the presence of polylysine, the average distance of the membrane-substrate interface was in the range of 35-40 nm. However, the cell membrane was highly protruded in the presence of other proteins like fibronectin, laminin or concanavalin-A. The presented method allows the nanoscopic characterization of the cell-sensor interface.

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Section: Discussionsupporting

confidence: 91%

Transmission electron microscopy study of the cell–sensor interface

Wrobel

Höller

Ingebrandt

et al. 2007

J. R. Soc. Interface.

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“…Adhesion contacts are defined as areas of closest distance between the membrane and the substrate (see Additional file 1 Figure S1). These findings contradict the work of Iwanaga et al [30]. For a more detailed study, we next created heat maps of the adhesion pattern ( Figure 4A) of each cell and investigated these adhesion patterns with regard to their spatial location ( Figure 4B), their number and their size ( Figure 4C).…”

Section: Adhesion Patch Distributioncontrasting

confidence: 72%

Cell membrane topology analysis by RICM enables marker-free adhesion strength quantification

et al. 2013

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Reflection interference contrast microscopy (RICM) allows the visualization of the cell's adhesion topology on substrates. Here it is applied as a new label-free method to measure adhesion forces between tumor cells and their substrate without any external manipulation, i.e., the application of force or adjustments in the substrate elasticity. Malignant cancer transformation is closely associated with the down-regulation of adhesion proteins and the consequent reduction of adhesion forces. By analyzing the size and distribution of adhesion patches from a benign and a malignant human pancreatic tumor cell line, we established a model for calculating the adhesion strength based on RICM images. Further, we could show that the cell's spread area does not necessarily scale with adhesion strength. Despite the larger projected cell area of the malignant cell line, adhesion strength was clearly reduced. This underscores the importance of adhesion patch analysis. The calculated force values were verified by microfluidic detachment assays. Static and dynamic RICM measurements produce numerous adhesion-related parameters from which characteristic cell signatures can be derived. Such a cellular fingerprint can refine the process of categorizing cell lines according to their grade of differentiation.

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“…[9,10] Several studies have been performed using physical techniques such as total internal reflection microscopy (TIRFM, [11] ) or fluorescence interference contrast (FLIC) microscopy in living cell cultures to measure the apparent distance of fibroblasts, rat brain astrocytes, embryonic rat hippocampus or snail neurons to oxidized silicon chip surfaces with terraces of 20 nm height differences. [12][13][14][15][16] Usually microscopic methods are used to analyze cell-to-material contacts especially when cell integration onto and into complex structures need to be visualized. [4,17,18] Conventional histological methods in combination with transmission electron microscopy (TEM) and scanning electron microscopy (SEM) are performed.…”

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confidence: 99%