[Cell Phenotyping] 포유류 세포
|Date : 2019.11.26|
|분류 : 포유류세포|
Phenotype MicroArrays for Mammalian Cells
Phenotype MicroArray Mammalian assays are cell-based assays used to investigate up to 1,400 metabolic and chemical sensitivity
phenotypes of mammalian cells. They use 96 well plates pre-loaded with carbon-energy and nitrogen substrates, ions,
hormones/cytokines and anti-cancer agents. Plates are inoculated with the cells of interest and Biolog’s proprietary redox dye.
In some wells, the cells are stimulated and in other wells inhibited. The generation of energy-rich NADH by the cells reduces the
redox dye and brings about a color change which is then read with Biolog’s OmniLog automated incubator-reader.
Phenotype MicroArray Mammalian Assays can be used for diverse research and applications including:
The Assay Portfolio for Mammalian Cells Includes the Following:
• PM-M11 - PM-M14 : Anti-Cancer Agents
HOW PHENOTYPE MICROARRAY TECHNOLOGY WORKS
DNA Microarrays and Proteomic Technologies allow scientists to detect genes or proteins that are coregulated and whose
patterns of change correlate with something important such as a disease state. However there is no assurance that these
changes are really significant to the cell. Phenotype MicroArrays are a complementary technology providing the needed
information at the cellular level.
Phenotype MicroArrays are preconfigured sets of phenotypic tests deployed on microplate panels. Each well of the array is
designed to test a different phenotype after inoculation with a standardized cell suspension, allowing simultaneous testing of
thousands of phenotypes in a single experiment.
Phenotype MicroArrays use Biolog’s patented redox technology, with cell respiration (NADH production) as a universal reporter.
If the phenotype is strongly “positive” in a well, the cells respire actively, reducing a tetrazolium dye and forming a strong color.
If it is weakly positive or negative, respiration is slowed or stopped, and the result is less color or no color.
The redox assay provides for both amplification and precise quantitation of phenotypes. Incubation and recording of phenotypic
data is performed automatically by the OmniLog instrument.
To compare the phenotypes of two cell lines, one is recorded as a red tracing and one as a green tracing. These graphs can
then be overlaid by the bioinformatic software to detect differences. Areas of overlap are colored yellow, whereas differences are
highlighted as patches of red or green.
KINETIC DATA CAPTURE AND ANALYSIS
OmniLog PM software contains a suite of algorithms that work in conjunction with the OmniLog PM system and
Phenotype MicroArray panels to automate incubation of up to fifty microplates at a fixed user-controlled temperature with
complete collection of colorimetric assay data over time. These programs allow for display of kinetic data from PM panels
recorded by the OmniLog PM system, manage and analyze the data, export it in a variety of raw and processed forms,
and generate reports.
• Drives the operation of the OmniLog PM system
• Guides loading and reading of plates or PM panels
• Creates a kinetic data file for each plate or PM panel for use in the other
File Management/Kinetic Analysis
• Assembles plate or PM panel data files into data lists
• Displays kinetic plots of the data
• Allows export of kinetic plots as bmp or jpeg files
• Extracts data lists from the File Management /Kinetic Analysis module
• Calculates parameters from kinetic data
• Allows comparison of two data lists
• Highlights wells and generates a report on PM phenotypes that differ
significantly in any selected kinetic analysis parameter
• Allows identification of substrates in all PM panels
• Links metabolic substrates in PM panels to KEGG database
• Export the original OmniLog kinetic dataExport data parameters for
statistical & bioinformatic analysis