Achieve accuracy and reliability in cell counting

Left to right: Yingxin Liang from the College of Pharmaceutical Sciences at Zhejiang University, Dr. Tianpeng Gu from the School of Medicine at Shanghai Jiao Tong University, and Weihao Liang from the Southern Medical University

We speak to three scientists from diverse research backgrounds to explore instrument features they consider crucial for achieving reproducible cell counting and viability assessments. We also learn how the reliability and speed offered by their preferred instrument - the Countstar Mira cell counter - sets it apart and helps them achieve an error-free and simplified laboratory workflow.

Tissue digest samples play a critical role in a variety of research and clinical applications. When handling these samples, accurate cell count and viability measurements post-sample preparation or before downstream cell-based assays, is essential to obtain reliable and comparable results in research. However, the complexity of tissue samples, which often contain multiple cell types of different shapes and sizes, along with cellular and matrix debris and cell clusters, poses challenges for most conventional automated image cell counters to analyze.

To address this issue, the team at ALIT BioTech - a company that specialises in advanced equipment and technology serving the life sciences sector - designed the Countstar Mira cell counter series. The Countstar range hosts specialized cell counting devices powered by AI-based image process algorithms to accurately identify and count living cells. We speak with three researchers studying diverse topics in the life sciences field to understand how the Mira FL fits into their labs' workflows: Yingxin Liang from the College of Pharmaceutical Sciences at Zhejiang University uses it to facilitate lung organoid research and culturing, Dr. Tianpeng Gu from the School of Medicine at Shanghai Jiao Tong University uses it for studying regulation of biological processes and stem cells involved in development, and Weihao Liang a PhD student in Southern Medical University and affiliated with the Nanfang Hospital, uses it to investigate blood-based cellular interactions.

Excellent cell identification

Highlighting the Mira FL's advantage in reliably identifying cells from complex sample types, Yingxin Liang explains, "My research on human lung organoids requires frequent assessments of the total number of live cells in tissue digests. In my field, cell counters that usually employ the Trypan Blue method misclassify organoid aggregates as dead cells. However, Mira FL's excellent cell-identification function, which leverages fluorescent labelling via the acridine orange and propidium iodide (AO/PI) staining method, precisely distinguishes between dead and living cells. This improves both, the accuracy, and the subsequent success rate of organoid culture."

Powerful dependability

Similarly, Dr. Tianpeng Gu - also an advocate of the accuracy of the Countstar Mira series - attributes the range's dependability to the powerful combination of parameter optimization and AI-driven processing. "My team studies the regulation of DNA methylation by DNA methyltransferases and TET dioxygenases, examining its biological role in development and in regulating stem cells," he states. "The Countstar Mira has been a staple in our cell counting experiments. We depend on its ability to achieve accurate cell counts by setting specific exclusion parameters, such as excluding multicellular clusters and cell fragments, at the start of experiment."

Enhanced counting accuracy

The Countstar Mira allows users to adjust the cell diameter setting to further include or exclude cells to be measured based on size, as well as identify and count individual cells within clusters. Dr. Tianpeng Gu says, "this parameter optimization coupled with the fact that the Countstar Mira counts cells across three to five fields of view and averages the numbers to get its results, helps optimize the reliability and reproducibility of our cell viability assessments." Yingxin Liang also agrees on the advantage that the AI-powered image processing feature bestows in cell counting experiments, as he notes that "the ability to recognize images through training the AI allows the Mira FL to identify and count specialized cell samples such as organoids more effectively than other cell counters."

Both Gu and Weihao Liang also express satisfaction at the Mira FL's ability to reliably determine cellular viability across different cell types - in stem cells for Gu, and in human primary neutrophils for Liang - using Trypan Blue Dye staining. "As the Mira can automatically determine cell viability concurrently with cell counting, it greatly enhances the counting accuracy," notes Gu.

Speedy analysis

Another advantage that Countstar Mira offers is that of speedy analysis. Weihao Liang says, "In my research - which focuses on the cellular interaction between platelets and neutrophils - given the rapid response and short lifespan of the cell types we study, having an accurate and timely cell counter is critical. Utilizing the Mira's advanced technology, especially the specialized platelet count module, we can now complete platelet counts for five samples at one stretch in just under two minutes. By reducing roughly 90% of the time we spend on cell counting, the Mira FL has enabled us to efficiently conduct experiments while preserving cell activity and accommodating sufficient sample numbers needed for independent analysis." Gu shares a similar sentiment, stating, "The Countstar Mira' ability to count five samples in less than one minute is a significant advantage compared to the hemocytometer we used prior to the Mira FL."

Streamlined lab operations

Overall, the enhanced accuracy and reproducibility that the Mira provides has helped all three researchers streamline their lab operations, while also eliminating sample errors for downstream assays. Yingxin Liang elaborates on the improvements in workflow efficiency since adopting the Mira FL in his lab, "Having used the Mira FL for a long time now to calculate the viability of primary cells isolated from tissue samples, the cell count results have helped me to continuously optimize our lab's primary cell isolation protocol," he asserts. "I have been able to develop an efficient method for isolating primary cells from human lung tissue, achieving over 90% viability of the isolated primary cells. This efficiency has been immensely helpful in culturing organoids."

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This release was published on openPR.