Cell Count and Viability for Advanced Flow Cytometry
Cell Count and Viability
Cell counting is a standard laboratory procedure that is routinely used within many experimental workflows, from life sciences to medical diagnostics. The assessment of cell counting in combination with viability is an important step in the characterization of cell health. Cell count and viability information can be used for monitoring proliferation rates, optimizing growth conditions and normalizing cell data for further studies.
In order to achieve optimum results, accurate and repeatable cell count and cell viability are essential. Traditional methods for measuring cell count and viability are often low throughput, lack linearity, and are time-consuming processes that do not provide the adequate event sampling necessary for reproducibility and accuracy of live cell analysis .
The iQue® Cell Count and Viability Kit provides accuracy across a large linear range in 96- and 384 well plates and reproducible analysis of absolute cell count and viability data from a variety of non-adherent cell lines with a streamlined workflow from cell labeling to analysis with no cell dilution required. Quantitate cell count and viability efficiently and accurately with iQue® Advanced High-Throughput Flow Cytometry!
iQue® Cell Count and Viability Kit Assay Concept
Designed for quantitative analysis of cell counts and viability, the assay is optimized to run on the iQue® platform with BR and VBR configurations, combining high throughput sampling and flow cytometry detection capabilities. It provides fast analysis of absolute cell count and viability across a large linear range from a variety of non-adherent cell lines with a streamlined workflow from cell labeling to analysis.
The kit includes validated reagents and pre-set templates for gating strategy and analysis. Using customer-provided cells, the kit identifies live cells and determines sample density in a quick, no-wash assay.
Basic assay workflow:
• Undiluted cells are mixed 1:1 with viability dye and incubated for 10-30 minutes
• Counting beads are added to the wells
• Plate is analyzed on the iQue® platform (BR and VBR configurations)
Increase accuracy and reproducibility
Ensure experimental reproducibility and accuracy to measure viability and absolute cell count
Perform high-throughput analysis
Quantify cell viability and count in 96 or 384-well plates across a large linear range without the need for sample dilution
Increase accuracy and reproducibility in biological workflows
Ensure robust experimental outcomes with multiplexed assessment of cell counting and viability
Figure 1. Reproducible and accurate viability and counts across a wide range
(A) Mixtures of growing and heat-killed cells (Jurkat, Raji, or Ramos cell lines) were tested at various ratios to ensure linear correlations throughout the range of sensitivity.
(B) Various densities of Jurkat cells were compared to determine linear range of cell counting. Undiluted cells were measured reliably from densities ranging from 8×104 to 2×107 cells/ml.
Perform high-throughput analysis
Quantify cell viability and count efficiently across a large linear range in 96 or 384-well plates
Figure 2. Heat and Profile Maps lead to actionable data
Limiting Dilution Cloning was performed by plating a limited dilution of CHO-S cells in a 96-well plate. Cells were incubated at 37°C, 5% humidity on an orbital shaker. Outer wells were media alone to prevent edge effects. Cell Density and Viability were determined using the iQue® Cell Count and Viability Kit. A Profile Map was generated to highlight wells that meet multiple criteria [(Cell Density > 0.3 million cells / ml) AND (Viability > 75%)]. Each metric in the profile map can be adjusted by dragging the slider bars at top of graph or by right-clicking to edit and the map of ‘hits’ will update automatically. The hits can be exported as a CSV file for hit picking or other downstream analysis.
(A) Heat map of cell density
(B) Heat map of viability
(C) Profile map showing hits matching higher density and viability
Save precious samples
Small sample size (10 µL) assures that precious samples are conserved for more critical downstream assays.
Figure 3. Easy to follow, low volume (10 µL/sample) protocol save cells and setup time
Maximize your productivity
Streamlined workflow with rapid, simple protocol, no washing, no cell dilution
Figure 4. Templated gating and analysis
Provided template with pre-set gating, acquisition settings, and analysis is imported into iQue® Forecyt software for quantitation of cell count and viability.