The in vitro addition of ATP finally triggered the continuous rotation of a few percentage of fluorescent actin filaments. These structures were mounted on cover glasses. fixed subcomplexes of F1 on surface-bound beads and attached a fluorescently labeled actin filament to each γ subunit of ATP synthase. To observe the rotation under a microscope, Yasuda et al. The analysis of such highly structured macromolecular complexes of sizes and dynamics within nanometer and microsecond ranges, respectively, requires preliminary knowledge about molecular players. On a purely molecular scale, imaging has for example provided an understanding of the rotational movement of F1-ATPase within ATP synthase. Optical molecular imaging can be used as a powerful tool for studying the temporal and spatial dynamics of biomolecules and their interactions, in vitro as well as in vivo. Molecular imaging is a discipline at the intersection between molecular biology and in vivo imaging. In many cases, however, the shown techniques can be used for a whole range of sample types. In the following sections we introduce biological systems ranging from single protein complexes to cell culture models and organisms of increasing complexity and give illustrative examples of appropriate light microscopy applications. Therefore, the selection of appropriate microscopes and data analysis tools requires the consideration of biological questions and sample properties (Figure 1). On the other hand, most microscopes are highly specialized devices. Furthermore, the growing palette of available fluorescent proteins and other fluorescent labels has facilitated the imaging of a broad range of sample types, ranging from single molecules to whole organisms. In addition, improved understanding of chemical and physical properties of genetically encoded fluorescence markers has led to the optimization of live cell imaging applications and limited undesired experimental side effects. Groundbreaking progress in technology during recent decades has leveraged the development of high-resolution microscopy. Light microscopy opportunities in systems biology In turn, the progress in imaging technologies requires computer vision techniques for automated image analysis. In addition, high-resolution three-dimensional (3D) imaging of small, whole organisms is now feasible across time. Recent progress in light microscopy allows for unprecedented insights into nanostructures, as well as unprecedented experimental throughput.
With regard to systems biology, microscopy is a tool that connects multiple scales of biological complexity, ranging from molecules to populations. Recent progress in biotechnology, technology, and interdisciplinary cooperation provides more realistic insights into biological processes than ever before. Current genomic tools in combination with advances in microscopy and computation facilitate in vivo observations of any genetic entity of interest. The main challenge of the post-genomic era is understanding the rules governing dynamic biological systems. However, understanding living dynamic biological systems by examining fixed specimens is, at the best, a heuristic process. For many decades, chemical fixation and the slicing of biological matter have been used to improve the stability and optical properties of samples. However, light scattering and other optical properties of living matter complicate the acquisition of informative images. One of the ultimate goals of systems biology is to elucidate relationships between molecular system states and higher order phenotypic traits.
Not surprisingly, visualization techniques are at the heart of science and engineering. Most of our sensory neocortex is engaged in the processing of visual inputs that we gather from our surroundings.
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