Once you have removed the cells from the disks, immerse the tip of the NucleoCassette into your sample, which aspirates a predefined volume of your sample into the cassette. The sample dissolves the propidium iodide dye, which has been immobilized inside the cassette. The cassette is then placed into the reading chamber of the NucleoCounter, activating the measurement cycle. A mechanical drive initiates the flow of sample into the counting chamber of the NucleoCassette, an image of the nuclei is recorded and analysis of the image occurs. In seconds, the count will be shown on the front panel display screen of the NucleoCounter. An optional, proprietary software program allows you to capture both an image and a numeric count for data logging purposes.
You can harvest cells, cell components, or intracellular protein by trypsinization, lysis buffer, freeze-and-thaw, sonication or any chemical or physical method. Specific protocols are available in New Brunswick Scientifics FibraStage Operating Manual.
Results and Summary:
Table 5, shown on the next page, provides data from a few of our recent studies using HEK-293, Sf-9 and CHO cells. These results were achieved using a single FibraStage bottle. For details of specific protocols, contact NBS. All results vary, and depend on the cell line, medium, nutrients and control parameters.
FibraStage System is a novel disposable bioreactor that is easy to use, is space-saving and labor-saving and achieves high cell densities compared to other 2-D bioreactors.
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