FCS Express: Chemometec Applications and Layouts

Chemometec

The Chemometec NucleoCounter NC-3000 is a full-spectrum fluorescence microscope which provides precise automated detection, quantification and analysis of fluorescence at the single cell level. Your multi parameter imaging data can now be easily analyzed in FCS Express and expanded upon using all of the tools and features FCS Express has to offer.

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Analysis Layouts

Click the links below to download some of the standard assay layouts for the NC-3000 system. If you do not already have FCS Express installed on your computer, please download and install the free FCS Express Reader before clicking on the links.

Application Layout Download Link
Apoptosis Download Apoptosis Layout
2-Step Cell Cycle Download Cell Cycle Layout
Caspase Download Caspase Layout
GFP Transfection Download GFP Transfection Layout
Mitochondrial Potential Download Mitochondrial Potential Layout
Vitality Download Vitality Layout
(Note: If you have a purchased copy of FCS Express, you can add your .ACS files to the Data List in any of the layouts above in order to view your own data)

Application Examples

Apoptosis

Download the PDF version of the layout: Chemometec Apoptosis Assay PDF or download and view the layout using the free FCS Express Reader: Chemometec Apoptosis Assay.

Assay Information:

Using fluorescence microscopy and image analysis the NC-3000 system automates detection of apoptotic cells based on phosphatidylserine externalization. Cells are stained with Hoechst 33342, PI and FITC labeled Annexin V. Hoechst 33342 stains the total cell population, while Annexin V stains apoptotic and necrotic cells. Early apoptotic cells exclude PI, while late stage apoptotic and necrotic cells stain positively for both Annexin V and PI.

2-step Cell Cycle

Download the PDF version of the layout: Chemometec Cell Cycle Assay PDF or download and view the layout using the free FCS Express Reader: Chemometec Cell Cycle Assay.

Assay Information:

Using fluorescence microscopy and image analysis, the NC-3000 system automates DNA content quantification and hence, measurements of cell cycle stages. The NC-3000 Two-Step Cell Cycle Assay uses the nuclear stain, DAPI, to measure DNA content. The protocol facilitates detaching, permeabilization, declumping and homogenous staining of the cell population in two simple steps without any trypsination, washing and centrifugation.

Caspase

Download the PDF version of the layout: Chemometec Caspase Assay PDF or download and view the layout using the free FCS Express Reader: Chemometec Caspase Assay.

Assay Information:

Using fluorescence microscopy and image analysis; the NucleoCounter NC-3000 system automates detection of apoptotic cells based on caspase activity. Cells are stained with Hoechst 33342, PI and carboxyfluorescein labeled FLICA reagent. The total cell population is stained with Hoechst 33342, while early apoptotic and late apoptotic/necrotic cells are stained with carboxyfluorescein labeled FLICA reagent and PI, respectively.

GFP Transfection

Download the PDF version of the layout: Chemometec GFP Transfection PDF or download and view the layout using the free FCS Express Reader: Chemometec GFP Transfection Assay.

Assay Information:

An easy way to determine the transfection efficiency is to use the gene encoding Green Fluorescent Protein (GFP) as a transfection reporter. The vector harbouring the gene of interest can be constructed so it also carries GFP either as a fusion or under control of its own promoter. Such a construct enables determination of transfection efficiency by simply determining the ratio of cells expressing GFP to the total cell number. The NucleoCounter NC-3000 enables two assays for measuring GFP transfection efficiency; a fast method taking less than a minute and an advanced assay which may be used for weak GFP expression and which also provides information about the transfection ratio of nonviable cells.

Mitochondrial Potential

Download the PDF version of the layout: Chemometec Mitochondrial Potential Assay PDF or download and view the layout using the free FCS Express Reader: Chemometec Mitochondrial Potential Assay.

Assay Information:

Using fluorescence microscopy and image analysis, the NC-3000 system automates detection of cells with collapsed mitochondrial membrane potential. Cells are stained with JC-1 and DAPI. Cellular JC-1 monomers and aggregates are detected as green and red fluorescence, respectively. Mitochondrial depolarization is revealed as a decrease in the red/green fluorescence intensity ratio. Necrotic and late apoptotic cells are detected as blue fluorescent (DAPI) cells.

Vitality

Download the PDF version of the layout: Chemometec Vitality PDF or download and view the layout using the free FCS Express Reader: Chemometec Vitality Assay.

Assay Information:

Using fluorescence microscopy and image analysis, the NC-3000 system automatically profiles the level of reduced thiols in a cell population. Cells are mixed with a solution containing VitaBright-48™, which stains viable cells in an intensity-dependent manner. Thus, a high fluorescence intensity of a particular cell indicates that the cell has a high level of thiols. A fluorescence intensity histogram showing the thiol level distribution in the cell population is automatically displayed after the analysis. By comparing histograms of treated cells to controls, the fraction of cells with low vitality (e.g. apoptotic or stressed cells) can be determined.