Scepter™ 2.0 Cell Counter Application Notes

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Discover Our Latest Cell Counting Application Notes Featuring Scepter™

To help you understand the potential of the Scepter™ Cell counter, we developed a series of Cell Counting Application Notes that explain the power of Scepter™ across a variety of cell lines.

Browse and review our applications notes summary and highlights or download them in PDF format.




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Identify CD4+ T cell differentiation toward Th2 and Th17 cell lineages

Identify CD4+ T cell differentiation toward Th2 and Th17 cell lineages
Using a 40 μm sensor, the Scepter cell counter enables the discrimination of cell types based on size with high resolution. The size distributions of Th2 and Th17 cell populations were compared to the size distribution of the CD4+ T cell progenitor cell type. Both Th2 and Th17 cells gradually increased in size from 6 to 10 μm after six days of differentiation.

T Cell Differentiation: Assessing CD4+ T Cells Differentiated Towards Effector T Helper Cell Lineages using Scepter™ 2.0 Handheld Automated Cell Counter

Murine CD4+ T cells can give rise to a variety of effector T, or "T helper", cell subsets depending on the nature of the immune response, and subsequently release a distinct subset of cytokines. We hypothesized that using the Scepter™ cell counter to rapidly assess size distributions of cellular populations would provide a quick, simple method for tracking T cell differentiation. As demonstrated in this study, Th1, Th2, and Th17 cells expanded from approximately 6 to 10 μm when compared to the progenitor CD4+ T cell type. This expansion upon differentiation was clearly, accurately, and precisely measured using the Scepter™ cell counter.

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Differentiate Lymphocytes and Monocytes

Differentiate Lymphocytes and Monocytes
We used the Scepter 2.0 cell counter to rapidly determine lymphocyte and monocyte concentrations as well as the relative frequency of these cell types in peripheral blood mononuclear cell (PBMC) isolates.

In each case, three main peaks were distinguishable by each analysis method in the separation of lymphocytes from debris. The peaks correspond to lymphocytes (small cells), monocytes (large cells), and a debris/dead cell fraction.

Immuno-monitoring: Immuno-monitoring Using the Scepter™ 2.0 Cell Counter and Software Pro

In addition to variations in protein expression, many immune cell types and physiological states are also uniquely distinguishable on the basis of size alone. Using the Scepter™ 2.0 cell counter’s sensitive size-discriminating capability, we demonstrated three examples of rapid, qualitative assessment of individual cell population frequencies in complex cell mixtures.

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Qualitative Cell Health Assessments

Qualitative Cell Health Assessments

Qualitative Cell Health Assessments
To examine the relationship between cell volume and camptothecin-induced apoptosis, we measured cell size distributions using the new Scepter handheld, automated cell counter.

In both cases, when measured with the Scepter Cell Counter, the treated cells showed a dramatic increase in the population of cells in the histogram to left of the main population. This indicated that there were larger populations of smaller size cells-likely representing the unhealthy cell populations.

Cell Death: Rapid Assessment of Size Changes Using the Scepter™ Cell Counter can be Predictive of Cell Death

It is a known phenomenon that cell volume changes occur early on in cell death pathways like apoptosis. Because the Scepter™ Cell Counter accurately measures cell volume and outputs histogram data by cell size, the instrument can be used to assess these morphological changes and thus be used as an early indicator of cells entering apoptosis.

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Count Cells in Milk

Count Cells in Milk
Somatic cells were purified from Dairy Milk using a simple and reliable spin wash protocol. The Scepter™ Data correlates well with data from the guava easyCyte which has the advantage of requiring no sample preparation. This data means the Scepter™ 2.0 Cell Counter can be used to accurately assess SCC in dairy cows.

Somatic Cell Counting: Rapid Counting of Somatic Cells in Dairy Milk Using the Scepter™ 2.0 Cell Counter

Mastitis is an inflammatory disease of the mammary glands in cows characterized by pathological changes in mammary tissue. Milk from sick cows exhibits increased somatic cell counts (SCC) due to the release of white blood cells into the gland to combat infection. Milk with high a SCC is of lower economic value than milk with low a SCC so it is crucial that SCC in monitored. This application note details the sample preparation required to get accurate counts using the Scepter™ 2.0 Cell Counter and correlates that data to the guava easyCyte.

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Assess Cell Cycle Changes

Assess Cell Cycle Changes
Size distribution profiles on the Scepter™ Cell Counter shift to the right when treated with the mitosis blocking agent Colchicine, indicating cells are larger as they accumulate in G2/M phase. The Scepter™ Cell Counter can be used as a cell culture monitoring device for a qualitative assessment of cell cycle disruption. More detailed analysis can be performed using Guava flow cytometer.

Cell Cycle: Novel and Rapid Method to Assess Proliferative Properties of Mammalian Cells by Monitoring Cell Size Distributions

In this study, we have used the Scepter™ Cell Counter to track cell volume/size changes as they relate to cell cycle, cell proliferation rate, and cell size. These results were compared to data from the guava flow cytometer, and we determined that the Scepter™ Cell Counter can be used for a rapid QC assessment of your cell culture by monitoring cell size shifts that occur during apoptosis and throughout cell cycle. More detailed analysis can be performed using Guava flow cytometer.

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Accurate Bead Counting

Accurate Bead Counting
Scepter™ counts beads with higher precision, as demonstrated by smaller coefficients of variation, compared to vision-based automated counting across multiple diverse bead types.

Bead Counting: Precise and Accurate Bead Counting Using the Scepter™ 2.0 Handheld Automated Cell Counter

Micron-sized beads are used in a variety of biological applications, ranging from daily validation of flow cytometer performance to purification of fusion protein constructs from cell lysates. Accurate determination of bead counts at the onset of each assay allows for standardization of bead concentrations across multiple samples and minimizes errors and variation in downstream results. Here we demonstrate how the Scepter™ cell counter is well suited for precise counting for beads of numerous types and can improve reproducibility of bead-based assays, such as immunoprecipitation and multiplexed detection.
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Separate human PBMCs from whole blood

Separate human PBMCs from whole blood
Our protocol for PBMC isolation enabled the generation of three distinct populations of blood cells, corresponding to RBCs, lymphocytes and monocytes. When the same PBMC samples were analyzed using flow cytometry (left) and a Scepter™ cell counter (right), the data showed close agreement between the analytical techniques.

Human PBMC Counting: Human PBMC Isolation and Counting Using the Scepter™ 2.0 Handheld Automated Cell Counter

Cellular assays using PBMC cultures form the backbone of immune monitoring studies in clinical diagnostics and therapeutic design. Given that ineffective separation of lymphocytes from whole blood can significantly alter cellular responses and lead to unreliable results, it is essential to start every assay with a rapid, simple, and reliable method of PBMC isolation and subsequent quantitation. We have shown that, by using the new 40 μm aperture sensor, the Scepter™ cell counter was able to accurately and precisely count a much broader range of cell types, including small cells (< 6 μm in diameter) such as PBMC and red blood cells (RBC). This application note outlines a protocol for the isolation of PBMC from whole blood and subsequent sample analysis using the Scepter™ cell counter.
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Count Yeast Cells during Fermentation

Count Yeast Cells during Fermentation
The Scepter™ cell counter counts yeast cells with good accuracy and linearity. Measured yeast cell concentrations were compared to theoretical concentrations. The solid gray line represents the theoretical values. Dotted lines represent best linear fit to data. Both the Scepter™ and Coulter Counter platforms show a loss of linearity and accuracy upon an increase in cell concentration.

Counting Yeast Cells: Counting Yeast Cells for Brewing and Wine Industries Can Be Facilitated by the Scepter™ 2.0 Handheld Automated Cell Counter

Adipocytes are derived from multipotent human mesenchymal stem cells (MSCs), providing researchers an ideal system for studying adipogenesis due to their multi-lineage differentiation potential. This study outlines a method for tracking adipogenic differentiation of ADSCs and 3T3-L1 cells and subsequent sample analysis using the Scepter™ cell counter to measure cell size and volume, as well as lipid vacuole staining using Oil Red O as a cross- validation histology screen to investigate the relationship between cell differentiation and cell size changes. Here we demonstrate how Scepter™ can also function as a reliable tool to track phenotypic change, in addition to generating highly precise cell counts.
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Preadipocytes (ADSCs) can be distinguished from differentiated adipocytes based on cell size

Preadipocytes (ADSCs) can be distinguished from differentiated adipocytes based on cell size
Using a 60 μm sensor, the Scepter™ cell counter enabled the discrimination of cell types based on size, with high resolution. ADSCs were measured at three key time points: Day 0 (control), seven (7) days after exposure to differentiation conditions, and fourteen (14) days differentiated. As indicated by the histogram data, cells gradually increased in size from 15 to 21 μm over the fourteen-day differentiation.

Adipogenesis Monitoring: Visualizing differentiation of adipocyte Using the Scepter™ 2.0 Cell Counter and Software Pro

Adipocytes are derived from multipotent human mesenchymal stem cells (MSCs), providing researchers an ideal system for studying adipogenesis due to their multi-lineage differentiation potential. This study outlines a method for tracking adipogenic differentiation of ADSCs and 3T3-L1 cells and subsequent sample analysis using the Scepter™ cell counter to measure cell size and volume, as well as lipid vacuole staining using Oil Red O as a cross- validation histology screen to investigate the relationship between cell differentiation and cell size changes. Here we demonstrate how Scepter™ can also function as a reliable tool to track phenotypic change, in addition to generating highly precise cell counts.
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