Atomic force microscopy – looking at mechanosensors on the cell surface

Heinisch, Jürgen J.; Lipke, Peter N.; Beaussart, Audrey; El-Kirat-Chatel, Sofiane; Duprès, Vincent; Alsteens, David; Dufrêne, Yves F.

doi:10.1242/jcs.106005

Cited by 43 publications

(43 citation statements)

References 59 publications

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“…In order to begin examining the ultrastructure of the C. albicans cell wall, we used atomic force microscopy (AFM), as it gives high-resolution images of the cell wall in a liquid environment without damaging the sample. This allows real-time imaging of metabolically active samples (50)(51)(52)(53). First, we needed to immobilize the cells on a suitable surface.…”

Section: Resultsmentioning

confidence: 99%

“…The cells are dispersed uniformly, making it easier to detect cells and image them by AFM. A lower percentage of gelatin (0.025%) was used, as it seemed to reduce the level of aggregation, which was higher with higher percentages of gelatin (i.e., 0.1 to 0.5%) (51).…”

Section: Resultsmentioning

confidence: 99%

“…In combination with other work on imaging and immobilization of cells (28,51,56), our approach to analyzing interactions between host lectins and pathogens using functionalized tips provides another method to address nanoscale interactions in fungal immunology. Exposure of ␤-(1,3)-glucan plays an important role in the detection of C. albicans by macrophage cells, and therapies to enhance exposure could potentially aid in the control and elimination of C. albicans infections.…”

Section: Discussionmentioning

confidence: 99%

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β-(1,3)-Glucan Unmasking in Some Candida albicans Mutants Correlates with Increases in Cell Wall Surface Roughness and Decreases in Cell Wall Elasticity

et al. 2017

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Candida albicans is among the most common human fungal pathogens, causing a broad range of infections, including life-threatening systemic infections. The cell wall of C. albicans is the interface between the fungus and the innate immune system. The cell wall is composed of an outer layer enriched in mannosylated glycoproteins (mannan) and an inner layer enriched in ␤-(1,3)-glucan and chitin. Detection of C. albicans by Dectin-1, a C-type signaling lectin specific for ␤-(1,3)-glucan, is important for the innate immune system to recognize systemic fungal infections. Increased exposure of ␤-(1,3)-glucan to the immune system occurs when the mannan layer is altered or removed in a process called unmasking. Nanoscale changes to the cell wall during unmasking were explored in live cells with atomic force microscopy (AFM). Two mutants, the cho1Δ/Δ and kre5Δ/Δ mutants, were selected as representatives that exhibit modest and strong unmasking, respectively. Comparisons of the cho1Δ/Δ and kre5Δ/Δ mutants to the wild type reveal morphological changes in their cell walls that correlate with decreases in cell wall elasticity. In addition, AFM tips functionalized with Dectin-1 revealed that the forces of binding of Dectin-1 to all of the strains were similar, but the frequency of binding was highest for the kre5Δ/Δ mutant, decreased for the cho1Δ/Δ mutant, and rare for the wild type. These data show that nanoscale changes in surface topology are correlated with increased Dectin-1 adhesion and decreased cell wall elasticity. AFM, using tips functionalized with immunologically relevant molecules, can map epitopes of the cell wall and increase our understanding of pathogen recognition by the immune system.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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β-(1,3)-Glucan Unmasking in Some Candida albicans Mutants Correlates with Increases in Cell Wall Surface Roughness and Decreases in Cell Wall Elasticity

et al. 2017

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“…One solution to this problem is to attach biological molecules onto the AFM tip for specific molecular recognition. However, tip functionalization is complex and time-consuming, accurate data collection and interpretation remain technically challenging, it is difficult to standardize the experimental process, and it is difficult to avoid tip alterations or contamination during scanning [44,45].…”

Section: Challenge and Outlookmentioning

confidence: 99%

Progress of AFM single-cell and single-molecule morphology imaging

Liu

et al. 2013

Chin. Sci. Bull.

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Atomic force microscopy (AFM) can probe single living cells and single native membrane proteins in natural fluid environments with label-free high spatial resolution. It has thus become an important tool for cellular and molecular biology that significantly complements traditional biochemical and biophysical techniques such as optical and electron microscopy and X-ray crystallography. Imaging surface topography is the primary application of AFM in the life sciences. Since the early 1990s, researchers have used AFM to investigate morphological features of living cells and native membrane proteins with impressive results. Steady improvements in AFM techniques for imaging soft biological samples have greatly expanded its applications. Based on the authors' own research in AFM imaging of living cell morphologies, a review of sample preparation procedures for single-cell and single-molecule imaging experiments is presented, along with a summary of recent progress in AFM imaging of living cells and native membrane proteins. Finally, the challenges of AFM high-resolution imaging at the single-cell and single-molecule levels are discussed.

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“…The main advantages of AFM for characterizing the surface of live cells are that it can be used in liquid environments and that it provides spatial resolution on the nanometer scale [6]. AFM has provided a wealth of novel insights in cell biology, including nanoscale organization and dynamics of cell membranes [7], ultra-structures and activities of organelles [8], and dynamic structures of proteins [9]. AFM has thus become an important tool in cell biology, significantly complementing optical and electron microscopy, and X-ray crystallography.…”

mentioning

confidence: 99%

Atomic force microscopy imaging of live mammalian cells

Liu

et al. 2013

Sci. China Life Sci.

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Atomic force microscopy (AFM) was used to examine the morphology of live mammalian adherent and suspended cells. Time-lapse AFM was used to record the locomotion dynamics of MCF-7 and Neuro-2a cells. When a MCF-7 cell retracted, many small sawtooth-like filopodia formed and reorganized, and the thickness of cellular lamellipodium increased as the retraction progressed. In elongated Neuro-2a cells, the cytoskeleton reorganized from an irregular to a parallel, linear morphology. Suspended mammalian cells were immobilized by method combining polydimethylsiloxane-fabricated wells with poly-L-lysine electrostatic adsorption. In this way, the morphology of a single live lymphoma cell was imaged by AFM. The experimental results can improve our understanding of cell locomotion and may lead to improved immobilization strategies.atomic force microscopy, cell locomotion, lamellipodia, cytoskeleton, polydimethylsiloxane Citation:Li M, Liu L Q, Xi N, et al. Atomic force microscopy imaging of live mammalian cells.

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Atomic force microscopy – looking at mechanosensors on the cell surface

Cited by 43 publications

References 59 publications

β-(1,3)-Glucan Unmasking in Some Candida albicans Mutants Correlates with Increases in Cell Wall Surface Roughness and Decreases in Cell Wall Elasticity

β-(1,3)-Glucan Unmasking in Some Candida albicans Mutants Correlates with Increases in Cell Wall Surface Roughness and Decreases in Cell Wall Elasticity

Progress of AFM single-cell and single-molecule morphology imaging

Atomic force microscopy imaging of live mammalian cells

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