Cells
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It is hard to imagine a more fundamental as well as complicated
biological object than a living cell. Currently, optical microscopy
techniques are the primary method for cell surface visualization,
with microscopic characteristics of cells traditionally used for
diagnosis and classification of cancers. AFM is also applied in
this field as a separate technique or in combination with optical
microscopy. In this field Atomic Force Microscopy features not only
high-resolution imaging of cellular structures below the optical
limit, that is quite "natural" for this method, but also
evaluation of micromechanical properties of the cell and ability
to monitor cell dynamics and processes running in it even in real
time. At present, no other microscopic techniques are able to provide
directly both structural information of a biological sample and
related functional information at such high spatial resolution.
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| (a) AFM snap image of cell division (Escherichia coli bacteria cell). Scan size 6.5x6.5 µm. |
(b) AFM height image of a monolayer aggregate of
Escherichia coli bacteria cell (55 cells). Mica substrate. Scan size
13x13 µm. |
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| Fig. 1. Tapping mode AFM image of bacteria
cell (a) and aggregate of cells (b) obtained using NSC18 probe. Image
courtesy of Prof. I. V. Yaminsky, MSU (Moscow). |
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Direct imaging of fixed or living cells and sub-cellular structures
provides important information on the architecture of the membrane,
organelles and cytoskeleton of cells. AFM offers unique opportunity
to image, localize and identify integral membrane proteins at the
surface of living cells. Although one couldn't avoid staining or
fluorescent labeling to mark the proteins of interest due to indiscriminative
nature of AFM probing relative to chemical composition and nature
of the objects, further improvements and extensions promise this
problem to be solved. The other important function of AFM is its
ability to examine mechanical properties of tiny objects. Information
about micromechanical properties is quite important for cellular
systems because it helps to understand cell architecture and it's
functions. Local elastic properties of a cell can be quantitatively
derived from the force versus distance (F-S) curves obtained at
fixed surface points using AFM. Spatial variations of mechanical
properties are also reflected in the image contrast helping to visualize
different cell components.
The capabilities of AFM in studies of cell are extremely broad
yet their practical realization is far from being routine and easy-of-use
method. Using AFM cells can be imaged directly but sample preparation
might require a fixation of the cells to a substrate. AFM study
of cells in most native physiological media such as aqueous solutions
is also a more difficult procedure than in air. The imaging of cells
due to their large dimensions and soft nature might takes some time
because scanning rates in studies of cells with the contact and
oscillatory modes are quite low (0.2 - 0.5 Hz). Larger size of cell
can also require a use of AFM scanner with large Z-range and also
probes in which tip is longer than those applied for studies of
flat objects. All these questions are under consideration by AFM
manufacturers and, hopefully, researchers will get most appropriate
microscopes in near future.
Another major AFM application in cell studies is real-time monitoring
of living cells dynamics, intercellular interactions and response
to internal and external perturbations. The main problem in monitoring
dynamic behavior of the cell is diminishing of perturbations induced
by a probe during scanning process as well as maintaining stable
environmental conditions for both temperature and pH value. Another
technical challenge is to make higher temporal resolution since
time to acquire full scan of a living cell often exceeds characteristic
alterations being happened in it.
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