16 ANALYTICAL AND LABORATORY EQUIPMENT
Cell imaging multi-mode microplate reader
Enabling optical microscopy and cell- based assays with a flexible new solution.
T
he cell is life at its most basic. It is used as a model system for the
study of organism structure, function and dysfunction. Myriad cell lines and primary cells isolated from organisms are used throughout the life sciences.
Tese cell lines are probed in many ways, but two laboratory methods are critical to most research programmes involving the use of cells: optical microscopy and cell-based assays.
Optical microscopy is used to perform routine laboratory tasks such as cell counting and determination of transfection efficiency. When used with stains, such as hematoxylin and eosin, optical microscopes can be used for histological applications useful for the diagnosis of cancer.
Fig. 1. On the left is an image of the Cytation 5 showing the open microplate tray. On the right is an artistic impression highlighting the phase contrast and fluorescence microscopy optics.
Equipped for fluorescence microscopy, a broad palette of fluorescent probes can be used, which include intercalating stains, reactive fluorescent dyes,
fluorescently-labelled antibodies and the family of GFP variants.
Tese probes allow for a number of experiments including localisation of quantification of DNA and proteins within the cell; conducting phenotypic assays based on protein translocation and cell morphology changes; and performing sub-population analysis of the cells in the field of view based on a phenotype expression (i.e. cell cycle). Furthermore, optical microscopy with phase contrast optics provides adequate contrast in cell monolayers and thin tissue sections such that label-free, live cell analysis is possible.
Cell-based assays typically quantify attributes of populations of cells. Tese assays are most often conducted in 96- or 384- well microplates with microplate readers. Tese readers use PMT detection and optical paths designed to capture as much light from the well as possible. Cell-based assays can quantify
ligand binding to receptors; the expression of second messengers; post-translational modifications of proteins, such as phosphorylation and ubiquitinylation; cellular viability, toxicity and apoptosis; and many other cellular attributes.
Multi-mode microplate readers provide many detection modes including the most common based on absorbance, fluorescence and luminescence detection.
Multiple instruments to conduct these tasks In some laboratories, perhaps as many as half a dozen different instruments are used to perform all the applications described above with optical microscopy and cell-based assays. Tese include: bright field microscope with oculars; colour-based digital microscope for histological staining; fluorescence microscope with oculars or monochrome CCD digital microscope; phase contrast microscope for
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