Chemometec - NucleoCounter英文圖文介紹
| The NucleoCounter is an instrument suited for the counting of cells and estimate of viability. Optionally it can be connected to a PC for documentation of results and images. The technique that the NucleoCounter is based on offers great advantages compared to existing methods. High precision, fast analysis, small sample consumption, simple operation, reliable operation. |
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| One of the inventive aspects of the NucleoCounter is the NucleoCassette. It is a disposable sampling and measurement unit, which comes pre-loaded with nuclei staining dye, Propidium-iodide. After analysis the NucleoCassette containing the sample, can be safely disposed of in the same manner as biological laboratory waste. |
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Heterogeneous cell lines such as adipocytes, CHO cells and hybridomas have successfully been measured without any adjustment of the instrument. The NucleoCounter is also well suited for the measurement of cells, which are grown on the surface of Cytodex Micro carriers. Samples that form cell aggregates or which are grown on the surface of the Cytodex microcarires are easily analysed. The sample pre-treatment involves the lysing of the cell plasma membrane thus dissolving any cell aggregates. This releases the cell nuclei to the solution and it becomes available for staining. |
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| The sample consumption of the NucleoCounter is very low. Volumes as small as 30 to 50 ul can be used for total cell count and volumes between 75 and 100 ul are needed for the non-viable cell count. These are the smallest usable volumes and due to dosage error it is generally recommended that volumes of between 200 and 400 ul are used. |
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| When estimating total count the sample pre-treatment involves the addition of two lysing buffers. Normally the buffers are added to the sample in volume equal to the sample volume. After the addition of the first buffer the sample is mixed for few seconds. The effect of the first buffer (Reagent A100) is the lysing of the cell plasma membrane. This lysing is completed virtually instantaneously. As soon as Reagent A100 has been mixed with the sample the second buffer (Reagent B) is added and the sample mixture thoroughly mixed again. The effect of Reagent B is partly to stabilise the nuclei but also to provide optimal conditions for the nuclei staining dye. The sample mixture is stable for considerable time after the addition of the reagents, it only needs to be mixed prior to analysis. |
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To analyse the sample it is loaded into the NucleoCassette where it dissolves the immobilised Propidium iodide nuclei staining dye. About 50 ul are loaded in the cassette by pressing the piston. When estimating the number of non-viable cells per volume the NucleoCassette is loaded with the sample without any sample pre-treatment. For the estimate of total count it is the sample/reagent mixture which is loaded into the cassette. |
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| When the sample or sample mixture has been loaded into the NucleoCassette it is ready to be analysed without further delay. The cassette is inserted into the NucleoCounter instrument where the final mixing of the sample or sample mixture and the nuclei staining dye takes place. The instrument needs no warming-up or preparation. It is ready to measure few seconds after it has been turned on. |
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When the NucleoCassette has been inserted into the NucleoCounter the operator simply has to press the "Run" button to initiate the analysis. The NucleoCounter performs the analysis in about 30 seconds and the resulting cell count is presented in the built-in display of the instrument. |
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| For visual inspection of the collected image and for documentation purposes the The NucleoView is a software which allows the user to generate and print tables with the results as well as doing viability calculations. |
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