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Abstract
Atomic force microscopy is a valuable and useful tool for the imaging and investigation of living cells in their natural environment at high resolution. Procedures applied to living cell preparation before measurements should be adapted individually for different kinds of cells and for the desired measurement technique. Different ways of cell immobilization, such as chemical fixation on the surface, entrapment in the pores of a membrane, or growing them directly on glass cover slips or on plastic substrates, result in the distortion or appearance of artifacts in atomic force microscopy images. Cell fixation allows the multiple use of samples and storage for a prolonged period; it also increases the resolution of imaging. Different atomic force microscopy modes are used for the imaging and analysis of living cells. The contact mode is the best for cell imaging because of high resolution, but it is usually based on the following: (i) image formation at low interaction force, (ii) low scanning speed, and (iii) usage of "soft," low resolution cantilevers. The tapping mode allows a cell to behave like a very solid material, and destructive shear forces are minimized, but imaging in liquid is difficult. The force spectroscopy mode is used for measuring the mechanical properties of cells; however, obtained results strongly depend on the cell fixation method. In this paper, the application of 3 atomic force microscopy modes including (i) contact, (ii) tapping, and (iii) force spectroscopy for the investigation of cells is described. The possibilities of cell preparation for the measurements, imaging, and determination of mechanical properties of cells are provided. The applicability of atomic force microscopy to diagnostics and other biomedical purposes is discussed.
Highlights
Introduction asYoung’s modulus and adhesion forces [6]
Studies on the a>]. TheAtomic force microscopy (AFM) force spectroscopy mode is used for the the mechanical properties of cells of different cancer determination of cell mechanical properties, such types show that the Young’s modulus of cancer cells is Correspondence to A
Principle of Atomic Force Microscopy Operation AFM can be applied for the investigation of surface morphology of cells at nanometric resolution under physiological conditions
Summary
(~400 μm in diameter) are connected to a vacuum source. A negative sucking pressure of 7–24 kPa (2–7 inHg) enables each through-hole to trap a single cell without damaging. The chemical fixation of cells simplifies the measurement process and improves image resolution. Chemical fixation dehydrates the cell and increases membrane stiffness; small components, such as cytoskeletal fibers, are not revealed in a topographical image [6]. Principle of Atomic Force Microscopy Operation AFM can be applied for the investigation of surface morphology of cells at nanometric resolution under physiological conditions. AFM is a label-free technique that can image the surface of living microbial cells at high resolution and practically “in real time,” thereby providing information that is complementary to that obtained by electron microscopy [8]. The electronics, which usually operate in a “closed loop mode,” control the probe movement in the z axis, according to the signal received from the detector This feedback circuit keeps a constant distance between the probe and the surface. The computer processes the “error signal,” i.e., the difference between the set force and the detector signal, and converts this computation into the image of topographical or other characteristics
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