Cells to find the rare outstanding performers (17 minutes at 1000 cells per
Cells to find the rare outstanding performers (17 minutes at 1000 cells per second). With the modern high speed sorters this will take less than a minute. In the following, utilization of cell sorting in biotechnology will therefore be highlighted with the main emphasis on AICAR custom synthesis fluorescence activated cell sorting. The actual selection of single cells is achieved by different types of sorters, the most frequently used ones being jet-in-air sorters. The liquid stream with the cells, after passing the laser light and the optics, is split up into defined droplets. The droplets containing a cell to be sorted is charged and then deflected into either a separate tube or directly into the individual wells of a microtiter plate. Table 1 provides an overview of typical applications of cell sorting in biotechnology.sorting allows a more in depth characterization of cells with specific properties observed in flow cytometric analyses, by sorting cells from different observed subpopulations. Subsequently, these cells are analyzed by other methods, thus linking different types of information and analytical methods to enhance the understanding of cell behavior. Cell viability is probably the most widely used parameter in this respect. A comprehensive study of viability assessment by flow cytometry and cell sorting has been described by Nebe-von Caron et al. [7]. By triple fluorochrome staining using propidium iodide, ethidium bromide and bis-oxonol, it is possible to discriminate between undamaged, damaged (membrane depolarized) and dead cells, which was verified by sorting and plating of the different subpopulations. Similar approaches have been followed for lactic acid PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/28607003 bacteria [8,9], further underlying the validity of fluorescent viability staining. ComasRiu and Vives-Rego extended this concept for Paenibacillus polymyxa by including the forward scatter signal into the assessment, thus discriminating between live and dead vegetative cells as well as viable and non-viable endospores [10]. Using a similar approach as described above for bacteria, the utility of flow cytometric viability assessment was verified for baker’s yeast by sorting and plating [11]. M ler and L che analyzed populations of brewing yeast for the content of DNA, neutral lipids and hydroxysterol by flow cytometry, verifying the data with cell sorting and image analysis [12]. Petit et al. described the use of cell sorting (combined with flow cytometry and confocal microscopy) for the study of respiratory dysfunction in yeast [13]. During the last years the distribution of cellular properties, as observed by flow cytometry, and the sorting of specific subpopulations have attracted additional attention for transcriptomic studies. Many biological samples are cell mixtures, and it was shown that sorting the differ-Physiological researchFlow cytometry, but also cell sorting have become valuable tools for physiological research in biotechnology. CellPage 3 of(page number not for citation purposes)Microbial Cell Factories 2006, 5:http://www.microbialcellfactories.com/content/5/1/ApIII pVIIIBOmpA OM PG PP CMCSpA PG CMDcell wall protein e.g. AgEAga2 AgaFCW CMCWCMgp64 nucleocapsid lipid envelopeCMFigure 2 Pro- and eukaryotic surface display systems Pro- and eukaryotic surface display systems. A: phage display, e.g. phage M13. pIII: minor capsid protein, pVIII: major capsid protein [19]. B: gram negative bacteria, e.g. E. coli. Anchor protein: OmpA, CM: cytoplasma membrane, OM: outer membrane, PP:.