It should be noted that by methods of color decomposition or by unique genetic labeling methods, such as the ‘brainbow’ method, it is possible to separate dozens of combinatorial labeling options ( Cachero and Jefferis, 2011 Livet et al., 2007). For example, microscopes employing current technologies can resolve up to six synthetic fluorescent colors in a fixed sample and up to three fluorescent colors in live samples. In order to take full advantage of HCS systems, experiments should be planned to maximize data content. Despite their sparsity, these readouts at the single-cell level are essential for assaying diversity within a population. These include flow cytometry, high-throughput microscopy set-ups, high-throughput single-cell sequencing and DNA methylation assays. These include plate readers (to study processes such as enzymatic function, protein–protein interactions and expression levels), deep and next-generation sequencing platforms (to study genomes and expression patterns), microarrays (expression chips, intron chips, chromatin immunoprecipitation, single nucleotide polymorphisms and genetic diversity), lipid arrays, protein arrays and whole-cell mass spectrometry (for proteins or metabolites).īy contrast, platforms that allow readouts at the single-cell level are less abundant to date. One of the main differences between these current technologies is whether they allow readouts on the level of the population or the single-cell.Ĭurrently, a number of platforms that allow readouts on the population level are available. Different types of such systematic high-throughput studies are geared towards quantifying and analyzing separate aspects of cell biology, such as genomics, transcriptomics and proteomics. Technological developments in the past few decades have led to a flourish of large-scale studies that have yielded a wealth of data. The repertoire of available high-throughput platforms coli developed at the National Institute of Genetics, Japan), and these could also be utilized for HCS.īox 1. Additionally, deletion libraries have been created for fission yeast ( Kim et al., 2010) and for bacteria (e.g. This can be done, for instance, by RNA interference (RNAi) in cell culture ( Brass et al., 2008 Krishnan et al., 2008 Moffat et al., 2006 Neumann et al., 2006 Prudencio et al., 2008) or by using a deletion library, as has been performed in budding yeast ( Vizeacoumar et al., 2010). For example, it is now feasible to perform functional genomics screens by capturing a microscopic phenotype of cells in which each gene has been knocked out or knocked down systematically. Combining the richness of visualization approaches with various experimental strategies creates nearly endless options for generating biological insights ( Fig. In addition, the technological advance of biological tools and microscopic platforms now enables screens to be performed in an array of genetic backgrounds, under different growth conditions and at various time points, as well as allowing the comparison of multiple tissues, cell lines or organisms. At the basis of this capability lies the ever-growing variety of labeling methods (discussed below) that allow visualization of cellular architecture and function, as well as developmental or behavioral processes ( Fig. High-throughput microscopy can be employed to address a wide spectrum of biological questions. Such advances are placing HCS methodologies at the frontier of high-throughput science and enable scientists to combine throughput with content to address a variety of cell biological questions. In this Commentary, we will discuss recent work, which has used HCS, to demonstrate the diversity of applications and technological solutions that are evolving in this field. These features make HCS a powerful method to create data that is rich and biologically meaningful without compromising systematic capabilities. ![]() ![]() Moreover, it allows the visualization of an enormous array of cellular features and provides tools to quantify a large number of parameters for each cell. High-throughput microscopy, also often referred to as high-content screening (HCS), allows acquisition of systematic data at the single-cell level. The ease with which data can now be generated has led to a new culture of high-throughput science, in which new types of biological questions can be asked and tackled in a systematic and unbiased manner. The increasing availability and performance of automated scientific equipment in the past decades have brought about a revolution in the biological sciences.
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