Cell Biology

Cell Count & Viability for Consistent Results

In cell biology research, controlling cell counting procedures at the earliest stage encourages reproducible results. Using an automated cell counter is recommended to ensure equal numbers of cells are used in downstream analysis, and to determine cell concentration in an accurate and precise manner.

Solutions

In cell biology research, whether you need cell count and viability, or to perform advanced fluorescence-based cell analysis with high reproducibility, we have a solution to suit your needs. Our instruments are automated fluorescence image cytometers designed for ease of use, precision and accuracy.

Obtaining high-quality research depends on your data’s reproducibility and consistency. NucleoCounter® instruments perform a wide range of functions, from precise cell count and viability determinations to advanced cell sample analysis, with minimal user interference.

Plug-and-play assays include five different apoptosis assays, a cell cycle analysis and a GFP-transfection assay. In addition, a platform for user-definable assays is also available. Our instruments are designed for dedicated cell biology research and development laboratories.

Unbiased Cell Count & Viability Determinations

Manual cell counting using a hemocytometer and trypan blue exclusion method is time-consuming and depends heavily on the user’s perception and pipetting skills.

To determine cell count and viability using the NucleoCounter® NC-202™ and NC-3000™ instruments, first load the cell suspension into the Via2-Cassette™ or Via1-Cassette™, respectively. Cells are automatically stained by the two fluorophores in the cassette, acridine orange and DAPI, staining total cells and dead cells, respectively. The volume of the counting chamber is pre-calibrated, ensuring high-precision and reproducibility.

High-speed Cell Viability & Counts

The multi-chamber NC-Slide A8™ enables high-speed viability and cell count determinations of insect and mammalian cells, measuring up to eight samples in less than three minutes. Mix the cell suspension with acridine orange and DAPI to stain all cells and the dead cells, respectively. Then, load the A8-slide™ chambers with the sample and insert the slide into the NucleoCounter® NC-3000™.

We offer optimized counting protocols for aggregated cells, cells growing on microcarriers and in spheroids. Read more about cell count and viability using the NC-Slide A8™ for mammalian cells.

Superior Data Visuals & Analysis

The NucleoView™ software is the user interface for the NucleoCounter® NC-200™ and NC-3000™. It handles data acquisition and presentation, along with image analysis. NucleoView™ includes a variety of data handling features that are perfectly suited to regulated environments.

The software enables you to visually inspect the fluorescence image and gives you the opportunity to verify the counting. You can select specific event populations in the scatter plots and examine and determine the validity of their inclusion or exclusion from the final counting results. Gating can be adjusted and you can save this adapted protocol for future measurements.

Counting Aggregated Cells

The NucleoCounter® NC-202™ (and NC-200™) includes a wide range of specialized assays making it the perfect cell counter. Sphere cultures or highly aggregated cell types pose great challenges in cell counting, but with our specialized assays, these instruments can count even the most aggregated cell culture samples.

Find out how you can get precise counts for aggregated cells.

Standardized Apoptosis Assays

For advanced research into cell death, it is essential to study the mechanisms of apoptosis. Using the NucleoCounter® NC-3000™, a series of five plug-and-play assays including Annexin V, mitochondrial potential with JC-1, caspase signaling, DNA fragmentation and the unique one-minute vitality assay, enabling you to fully investigate cell death mechanisms.

APOPTOSIS ASSAY PHYSIOLOGICAL CHANGES DETECTED STAGE
Mitochondrial potential Assay (JC-1) Collapse of the mitochondrial membrane potential Early
Annexin V Assay Collapse of plasma membrane lipid asymmetry Early-mid
Caspase Assay Caspase activation signals downstream apoptotic events Early-mid
Vitality (VB48™) Assay Decrease in cellular levels of reduced thiols e.g. GSH Late
DNA Fragmentation Assay Break-down and fragmentation of DNA Late

 

Covering early to late stages of apoptosis, these assays help you to carry out in-depth studies of apoptosis progression in mammalian cells. You can perform advanced data analysis effortlessly with NucleoView™. The platform presents links between images, histograms and scatter plots, giving you access to detailed and precise data analysis that identifies percentages of apoptotic, necrotic and living cells.

Monitoring Viability & GFP Transfection Efficiency

When transfecting cells with genes of interest, one way to determine the transfection efficiency is to co-express a fluorescent protein, such as green fluorescent protein (GFP) under the same promoter and determine the amount of GFP.

The NucleoCounter® NC-3000™ offers a fast and easy to use assay to test GFP transfection efficiency and expression levels. By staining cells with Hoechst 33342 and propidium iodide (PI), you can define the total cell population and the dead cell population together with the population expressing GFP.

Cell staining in NucleoView™ software using GFP transfection efficiency assay with the NucleoCounter® NC-3000™

A – Cells are located using Hoechst 33342 (blue) and the percentage of GFP expressing cells (green) can easily be determined. Non-viable cells are stained with propidium iodide (PI; red).
B – All cells stained with Hoechst 33342 (blue) are identified.
C – GFP expressing cells are identified in green and non-viable cells in red by PI staining.

Easy coupling between the image obtained and the scatter plots or histograms allow you to determine the precise gate settings used to investigate the staining pattern for specific cell populations. The NucleoCounter® NC-3000™ offers an all-in-one platform for evaluating GFP expression efficiency.

Non-viable cells stained with PI can be easily located with the image overlay function in the NucleoView™ software (A). GFP-transfected cells can be immediately identified (B).

Fast Cell Cycle Analysis

Investigating the impact of a treatment on cell division is one of the most powerful tools within cell biology. The NucleoCounter® NC-3000™ provides fast and easy cell cycle analysis in under five minutes.

After adding a lysis buffer, all cell nuclei are stained, and the sample is measured using the NC-3000™. A cell cycle profile displays in the accompanying Plot Manager in the NucleoView™ software. Events in the sub-G1-phase, G0/G1-phase, S-phase and G2/M-phase are identified.

With the FlexiCyte™ software package, the NucleoCounter® NC-3000™ can even be used to study cell proliferation with BrdU and EdU incorporation, detected with fluorescently labeled antibodies allowing for advanced studies of cell proliferation.

Illustration: Two-step cell cycle assay of untreated and camptothecin-treated (CPT) Jurkat cells. The histograms display intensity of the DNA-stain DAPI and can be used to define cell cycle events in the sub-G1-phase, G0/G1-phase, S-phase and G2/M-phase. After CPT treatment, the cell cycle is arrested in the G2/M-phase.

Advanced Cell Analysis

The FlexiCyte™ module available for the NucleoCounter® NC-3000™ enables users to perform detailed advanced cell analysis of their own choice in mammalian and insect cells. The combination of LEDs, from UV to far red, together with a carefully chosen set of emission filters, allows you to detect a broad range of fluorescent antibodies and proteins.

The Protocol Adaptation Wizard feature guides the user through a selection of optimal settings. After image acquisition, in the Plot Manager, cell data is presented beside the fluorescent image as either scatter plots, histograms, or both. By linking images with plots, you are armed with everything you need to perform detailed data analyses.

The FlexiCyte™ software package enables detailed biomarker analysis of a broad range of fluorescent antibodies and proteins.

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