233 research outputs found
Probing nanomechanical properties from biomolecules to living cells
Atomic force microscopy is being increasingly used to explore the physical properties of biological structures. This technique involves the application of a force to the sample and a monitoring of the ensuing deformation process. The available experimental setups can be broadly divided into two categories, one of which involves a stretching and the other an indentation of the organic materials. In this review, we will focus on the indentation technique and will illustrate its application to biological materials with examples that range from single molecules to living cell
Probing nanomechanical properties from biomolecules to living cells
Atomic force microscopy is being increasingly used to explore the physical properties of biological structures. This technique involves the application of a force to the sample and a monitoring of the ensuing deformation process. The available experimental setups can be broadly divided into two categories, one of which involves a stretching and the other an indentation of the organic materials. In this review, we will focus on the indentation technique and will illustrate its application to biological materials with examples that range from single molecules to living cells
Nanomechanical sensor applied to blood culture pellets: a fast approach to determine the antibiotic susceptibility against agents of bloodstream infections.
The management of bloodstream infection, a life-threatening disease, largely relies on early detection of infecting microorganisms and accurate determination of their antibiotic susceptibility to reduce both mortality and morbidity. Recently we developed a new technique based on atomic force microscopy capable of detecting movements of biologic samples at the nanoscale. Such sensor is able to monitor the response of bacteria to antibiotic's pressure, allowing a fast and versatile susceptibility test. Furthermore, rapid preparation of a bacterial pellet from a positive blood culture can improve downstream characterization of the recovered pathogen as a result of the increased bacterial concentration obtained.
Using artificially inoculated blood cultures, we combined these two innovative procedures and validated them in double-blind experiments to determine the susceptibility and resistance of Escherichia coli strains (ATCC 25933 as susceptible and a characterized clinical isolate as resistant strain) towards a selection of antibiotics commonly used in clinical settings.
On the basis of the variance of the sensor movements, we were able to positively discriminate the resistant from the susceptible E. coli strains in 16 of 17 blindly investigated cases. Furthermore, we defined a variance change threshold of 60% that discriminates susceptible from resistant strains.
By combining the nanomotion sensor with the rapid preparation method of blood culture pellets, we obtained an innovative, rapid and relatively accurate method for antibiotic susceptibility test directly from positive blood culture bottles, without the need for bacterial subculture
The Dynamics of Single-Cell Nanomotion Behaviour of Saccharomyces cerevisiae in a Microfluidic Chip for Rapid Antifungal Susceptibility Testing
The fast emergence of multi-resistant pathogenic yeasts is caused by the extensive-and sometimes unnecessary-use of broad-spectrum antimicrobial drugs. To rationalise the use of broad-spectrum antifungals, it is essential to have a rapid and sensitive system to identify the most appropriate drug. Here, we developed a microfluidic chip to apply the recently developed optical nanomotion detection (ONMD) method as a rapid antifungal susceptibility test. The microfluidic chip contains no-flow yeast imaging chambers in which the growth medium can be replaced by an antifungal solution without disturbing the nanomotion of the cells in the imaging chamber. This allows for recording the cellular nanomotion of the same cells at regular time intervals of a few minutes before and throughout the treatment with an antifungal. Hence, the real-time response of individual cells to a killing compound can be quantified. In this way, this killing rate provides a new measure to rapidly assess the susceptibility of a specific antifungal. It also permits the determination of the ratio of antifungal resistant versus sensitive cells in a population
Probing nanomechanical properties from biomolecules to living cells.
Atomic force microscopy is being increasingly used to explore the physical properties of biological structures. This technique involves the application of a force to the sample and a monitoring of the ensuing deformation process. The available experimental setups can be broadly divided into two categories, one of which involves a stretching and the other an indentation of the organic materials. In this review, we will focus on the indentation technique and will illustrate its application to biological materials with examples that range from single molecules to living cells
Nano-Motion Analysis for Rapid and Label Free Assessing of Cancer Cell Sensitivity to Chemotherapeutics.
Background and Objectives: Optimization of chemotherapy is crucial for cancer patients. Timely and costly efficient treatments are emerging due to the increasing incidence of cancer worldwide. Here, we present a methodology of nano-motion analysis that could be developed to serve as a screening tool able to determine the best chemotherapy option for a particular patient within hours. Materials and Methods: Three different human cancer cell lines and their multidrug resistant (MDR) counterparts were analyzed with an atomic force microscope (AFM) using tipless cantilevers to adhere the cells and monitor their nano-motions. Results: The cells exposed to doxorubicin (DOX) differentially responded due to their sensitivity to this chemotherapeutic. The death of sensitive cells corresponding to the drop in signal variance occurred in less than 2 h after DOX application, while MDR cells continued to move, even showing an increase in signal variance. Conclusions: Nano-motion sensing can be developed as a screening tool that will allow simple, inexpensive and quick testing of different chemotherapeutics for each cancer patient. Further investigations on patient-derived tumor cells should confirm the method's applicability
Yeast Nanometric Scale Oscillations Highlights Fibronectin Induced Changes in C. albicans
Yeast resistance to antifungal drugs is a major public health issue. Fungal adhesion onto the host mucosal surface is still a partially unknown phenomenon that is modulated by several actors among which fibronectin plays an important role. Targeting the yeast adhesion onto the mucosal surface could lead to potentially highly efficient treatments. In this work, we explored the effect of fibronectin on the nanomotion pattern of different Candida albicans strains by atomic force microscopy (AFM)-based nanomotion detection and correlated the cellular oscillations to the yeast adhesion onto epithelial cells. Preliminary results demonstrate that strongly adhering strains reduce their nanomotion activity upon fibronectin exposure whereas low adhering Candida remain unaffected. These results open novel avenues to explore cellular reactions upon exposure to stimulating agents and possibly to monitor in a rapid and simple manner adhesive properties of C. albicans
Simple optical nanomotion method for single-bacterium viability and antibiotic response testing.
Antibiotic resistance is nowadays a major public health issue. Rapid antimicrobial susceptibility tests (AST) are one of the options to fight this deadly threat. Performing AST with single-cell sensitivity that is rapid, cheap, and widely accessible, is challenging. Recent studies demonstrated that monitoring bacterial nanomotion by using atomic force microscopy (AFM) upon exposure to antibiotics constitutes a rapid and highly efficient AST. Here, we present a nanomotion detection method based on optical microscopy for testing bacterial viability. This novel technique only requires a very basic microfluidic analysis chamber, and an optical microscope equipped with a camera or a mobile phone. No attachment of the microorganisms is needed, nor are specific bacterial stains or markers. This single-cell technique was successfully tested to obtain AST for motile, nonmotile, gram-positive, and gram-negative bacteria. The simplicity and efficiency of the method make it a game-changer in the field of rapid AST
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