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Several surface spectroscopic and microscopic techniques have been used to characterize the surface in term of chemical composition and polymer topography.
The presence of antifreeze proteins or ice nucleating proteins in hemolymph is analyzed by various osmometric techniques or differential scanning calorimetry [6], [7], and electron and optical microscopic techniques have been used in some studies to elucidate ultrastructural details of cryodamage [8], [9].
New super-resolution microscopic techniques have begun to visualize internal mitochondrial membranes [ 46, 47] but are still incapable of rapid three-dimensional timelapse visualizations in live cells.
To further explore biological phenomena, two distinct microscopic techniques have been attempted to break the diffraction limit [ 6– 23] and have already been applied to live-cell imaging [ 20– 23].
In vivo microscopy in humans is still largely in the early stages, but various microscopic techniques have clinical potential, e.g. for the identification of suspicious tissue regions for biopsy collection (Liu et al., 2011).
An exception is the well studied chemotaxis pathway, where advanced microscopic techniques have revealed that proteins self-assemble stochastically in clusters whose function has only been theorized [ 2, 3].
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The potential application of monitoring the microcirculation in pediatric patients using hand-held intravital microscopic techniques has been exemplified in the work of Top and Tibboel.
Detailed critiques of this in vivo microscopic technique have previously been reported [ 6, 22- 27, 27, 36, 37].
In the past, Raman micro-spectroscopy as a label-free microscopic technique has been widely used because it offers high spatial resolution and high chemical specificity [ 10– 10].
To achieve this objective, several different and complementary microscopic observation techniques have been performed: optical microscopy, Scanning Electron Microscopy (SEM) and Atomic Force Microscopy observations and Electron BackScatter Diffraction (EBSD) measurements have been carried out on the surface of several specimens after cyclic loading.
A variety of microscopic imaging techniques have long been used to study chromatin architecture and nuclear organization [ 1- 3].
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