APPLICATION OF MICROSCOPY IN BIOSCIENCES

Application of Microscopy in Biosciences



Introduction

High content images are quickly becoming a mainstay in pharmaceutical science research and life laboratories.1 The power of the image lies in its ability to measure not only changes in fluorescence intensity in cells, but also proximal and temporal relationships and morphological characteristics. In recent years, the availability of a new generation of platforms for automated imaging has created a unique opportunity to develop new cell-based assays. High content microscopy has been overtaken by the abundant use of antibodies in immunofluorescence applications. These reagents have allowed high-resolution localization of important molecular targets in individual cells. For example, antibodies can be used to detect certain stages of apoptosis, the activation of signalling cascades, the organization or reorganization of cellular structures and the identification of cell cycle stages. However, many of the currently available antibodies have not been tested specifically for use in automated imaging applications. In addition, the potential for detecting high multiplexing is based on cell information. The ability to do this, however, is often limited to standard using primary antibodies labelled with labelled secondary antibodies cross-reactive species.

The STM was just the beginning of a now large family of scanning probe microscopes. Related instruments include atomic force microscopy, which allows for imaging and manipulation of electrically conductive as well as insulating material (e.g., living cells or single biomolecules in solution) at unprecedented resolution, as well as investigation of Micromechanical parameters such as stiffness and viscoelastic forces; scanning near-field optical microscopy; and scanning capacitance microscopy.

Besides these important and powerful techniques, various linear and nonlinear spectroscopy techniques are used to provide data. Electron and atom diffraction methods yield important additional information concerning the coherence of the investigated Microstructures. Electronic properties of Microstructures are often investigated via conventional emission methods such as electron, X-ray, or optical spectroscopy.

The optical methods comprise optical microscopy, confocal microscopy, X-ray microscopy, ultraviolet-visible (UV/VIS) spectroscopy, infrared spectrometry, terahertz spectroscopy, Raman spectroscopy, and surface enhanced Raman spectroscopy. Electron microscopy includes scanning electron microscopy and transmission electron microscopy. Additional methods include point-projection microscopes, low-energy electron diffraction, reflection high-energy electron diffraction, X-ray spectroscopy and diffraction, nuclear magnetic resonance, electron paramagnetic resonance, Auger electron spectroscopy and Mössbauer spectroscopy.

Microfabrication Tools

Microfabrication tools include electron beam lithography systems, sputter coaters, scanning electron microscopy (SEM) and SEM e-beam for maskmaking; resist spinners, Microimprint systems, contact aligners, (programmable) ovens and ultraviolet (UV) cure systems for optical photolithography, tools for chemical vapor deposition (low pressure and plasma enhanced), atomic layer deposition systems for epitaxial growth, evaporators and sputter systems for metallization and sputtering, plasma etchers and reactive ion etchers for dry etching, furnaces and rapid thermal annealers for annealing, oxidation, and doping, and tools for wafer bonding and sawing.

The large number of tools currently used for characterization, manipulation and fabrication in Microtechnology shows that there is “no best technique.” Since Microtechnology itself is at the meeting point of biology, engineering, chemistry, physics, biology, materials science, among others, its methods are also ...