How to validate an automatic cell counter: benefits of using validation slides

keyword: Automatic cell counter, cell counting accuracy, cell counter validation slides, ISO 20391 standard, cell counting reference materials, cell counting techniques, LUNA cell counter series, cell counting validation, cell viability assessment

1 Introduction

Automatic cell counters are widely popular for their ability to quickly and accurately provide cell counting results. Despite these advantages, verifying its accuracy is crucial to ensure reliable cell counting results. Microspheres are often used to validate devices such as flow cytometers and automatic cell counters.butThere are various issues with using microspheres, which has prompted the development of cell counter validation slides for verificationDian ChengLUNA ™ cell counterSeries.

In this article, we will explore cell counting standards and the use of a cell counter to validate slidesDian ChengLUNA ™ The advantages of the cell counter series.

2 International Cell Counting Standards

The development of ISO 20391 standard is aimed at providing guidance for accurate cell counting and ensuring the reliability of cell counting results. This standard outlines a scheme for verifying the quality of cell counting measurements through a series of dilution experiments, which can be used when there is no reference material available. However, this process may be time-consuming and complex, requiring complex calculations to determine the coefficients of determination and proportionality.

To overcome this challenge, it is recommended to use reference materials to ensure traceability of measurements, facilitate comparison, and validate the measurement process. An effective method is to compare the average analysis results with reference values obtained from proven reference materials, thereby eliminating the need for complex calculations. However, selecting appropriate reference materials still faces challenges.

The challenge of using validated microspheres as reference materials

Validation microspheres, such as those from the National Institute of Standards and Technology (NIST) in the United States, are often used as reference materials in various analytical methods involving particle size and counting. These microspheres have known sizes and concentrations and are often used as calibration standards for flow cytometry, microscopy, and other similar techniques. However, NIST microspheres do not have a fluorescent form and are not suitable for testing cell viability.

Although commercial fluorescent microspheres can serve as a substitute, errors during the pipetting process and evaporation during long-term storage may affect their accuracy, which requires careful handling and repeated testing. Therefore, there are currently no commercial reference materials available for evaluating cell viability.

Users who choose to prepare reference materials internally will have to rely on manual counting, which is relatively inaccurate and requires a large number of samples. Alternatively, flow cytometry can be used, although obtaining counting results may be challenging and not very user-friendly. Therefore, an alternative method for assessing cell viability needs to be found.

4. Use validation slides as reference materials

The use of stable and consistent reference materials is crucial for ensuring accurate results in cell counting. One option that can provide this level of quality control is to use validation slides. These glass slides contain pre spotted patterns or pre fixed fluorescent microspheres with a known number of objects on them, eliminating issues such as concentration changes caused by sample loading, evaporation during storage, or uneven distribution.

In addition, the validation slides allow the use of bright field and fluorescence optical systems of a cell counter to verify total cell count and cell viability. Cross validation with other devices is also simple, and its stability ensures that the measured values remain consistent even after long-term storage or repeated use. By using validation slides, researchers can save time and resources that were originally used for conducting additional experiments to ensure accuracy.

（A） Cell counter validation slide - Bright Field II (B) Cell counter validation slide - Fluorescent type

How to use a cell counter to verify the accuracy of slide evaluation

The validation slide scheme of the cell counter simplifies and improves the process of verifying the accuracy of the cell counter by providing a simple and efficient alternative to microspheres. This method saves time and provides a convenient way to verify the accuracy of cell counting results.

To verify the slide protocol using a cell counter, please follow these simple steps:

1) Check the cell counter to verify the cell concentration and viability values on the glass slide label.

2) Insert the glass slide into the cell counting device.

3) Start counting using the 'IQOQ' scheme.

4) Record the total cell concentration and cell viability values.

Automatic cell counter: More accurate counting brings higher quality science

Diancheng Biotechnology provides you with a variety of high-precision fully automatic cell counters, including bright field cell counters, fluorescent cell counters, automatic bacterial counters, as well as a comprehensive range of consumables and accessories. If you have any related needs, please feel free to contact us!