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Cell Counter Selection Guide: 4 Critical Criteria Every Lab Must Validate

   |  April 24, 2025

Using a cell counter can significantly improve the efficiency of cell counting experiments. However, certain challenges may arise when handling complex samples:

-Sampling Bias Risk:Single-field counts prone to cell distribution bias. Mitigate by mixing samples and averaging triplicate counts.

-Autofocus Limitations:Performance drops in low-concentration/heterogeneous samples, but stable with moderate density and uniform morphology.

-Fluorescence Detection Gaps:Single-excitation systems underestimate dual-fluorescence markers, restricting multi-parametric analysis despite cost savings.

For researchers, the accuracy of a cell counter is typically evaluated using the following three criteria:

-Double check the identification by the devices with Compare the Original Cell Image with the Instrument’s Detection Overlay: Assess the precision of cell recognition by checking how accurately the device outlines the cells in the captured image.

-Conduct Repeated Counting Experiments to Evaluate Inter-Group CV (Coefficient of Variation): Perform multiple replicate measurements to determine the consistency and reproducibility of the counts.

-Compare Results with Flow Cytometry or Manual Counting: Validate the instrument’s performance by benchmarking its results against those obtained from flow cytometry or traditional manual counting methods.

To enhance cell counting accuracy, the RWD C200FL Fluorescent cell counter was developed with a strong focus on three critical performance dimensions: imaging stability, image clarity, and algorithmic analysis capability. Each aspect underwent extensive optimization and technological upgrades.

Curious about the breakthrough core technologies behind these improvements? Keep reading — the details might just surprise you.

Field Imaging to Minimize Systematic Error

To address the limitations of single-field imaging and reduce systematic error, the C200FL Cell Counter is equipped with a 3-field counting mode. This design significantly expands the counting area, reaching a total of 7.6 mm². By distributing the fields across different regions of the sample, this approach minimizes sampling bias and provides a more accurate representation of cell distribution patterns.

Cell Counter Selection Guide
Cell Counter Selection Guide
CHO Cell Counting Linearity(R²>0.999)
Cell Counter Selection Guide
Cell Counter Selection Guide
High Repeatability in Multiple Counts of 293 Cells(CV<5%)

Fixed Focal Design Enhances Imaging Stability

The C200FL employs a fixed focal length imaging system, which minimizes fluctuations caused by autofocus algorithms and significantly reduces counting time. This design ensures consistent image quality across multiple captures, thereby lowering the coefficient of variation (CV) in repeated measurements. Clear and stable imaging provides a reliable foundation for accurate cell recognition by the analysis algorithm.

Cell Counter Selection Guide
Cell Counter Selection Guide

Intelligent Algorithm Enhances Cell Recognition Accuracy

In addition to conventional parameters such as diameter, circularity, and fluorescence thresholds, we have implemented advanced cell boundary interpretation parameters. This allows for precise identification of cell edges, improving the accuracy of detecting clustered cells and enhancing overall cell recognition performance.

Cell Counter Selection Guide
Note: The yellow circle indicates cell clusters.

Highly Sensitive Fluorescence Detection System for Broader Applications

The RWD cell counter series features an independent dual-fluorescence channel optical design. The C200FL model is equipped with high-performance optical components and background purification algorithms, enabling it to effectively capture weak fluorescence signals while minimizing background noise. This results in clear, noise-free fluorescence images. In comparison tests with flow cytometry, the transfection rate detected by both systems differed by no more than 10%.

Cell Counter Selection Guide
Peripheral Blood PBMC Cell AOPI Fluorescence Counting (Green represents live cells stained with AO fluorescence, while red indicates dead cells stained with PI fluorescence.)
Cell Counter Selection Guide
Cell Counter Selection Guide
Cell Counter Selection Guide
Cell Counter Selection Guide

RWD C200FL Fluorescent Cell Counter

Cell Counter Selection Guide

With core technologies such as multi-field imaging, high-performance optical design, and precise cell recognition algorithms, the C200FL Fluorescent Cell Counter overcomes the limitations of traditional cell counters. It provides researchers with efficient and accurate cell counting solutions, meeting the industry’s demand for high-precision data. Additionally, its software design complies with FDA 21 CFR PART 11 requirements, ensuring that corporate users’ needs for electronic records and access control are fully met.

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