Abstract

The tedious and time-consuming process of manually counting cells on a hemocytometer has been greatly alleviated in recent yearswith the advent of various automated cell-counting instruments. Even with these advancements, the measurement of cell concentration and viability for many samples is a common bottleneck in large scale experiments and many industrial processes.For example, single-sample automated cell counters may not be appropriate for counting multiple primary murine samples after a large take down and during cell line development and manufacturing. To address these challenges, we have developed the CellacaMX a high-throughput automated cell counting system. The system can image, analyze, and report cell concentration and viabilityfor 24 samples in 48 seconds using bright field (trypan blue) and in 2.5 minutes using multiple fluorescent imaging channels. The instrument was developed to operate in both a manual mode and as a fully automated, plate-based system, thus providing the user greater set-up flexibility depending on the project and sample quantity. In addition, small loading volume (50 –200 μL per sample) assures that precious samples are conserved for more critical downstream assays. We present a comparison between counts obtained on the CellacaMX instrument and those obtained using the laborious gold-standard hemocytometer method. We further tested the platform using CHO cells stained with trypan blue, routinely used in bioprocessing. Finally, we share resultsof an AOPI cell viability assay on Jurkatcells. Our experiments demonstrate a cell-counting system capable of increasing both accuracy and throughput in biological workflows. This advancement is of significant value to the cell line development and bioprocessing communities. It provides an efficient method of counting and analysis of multiple samples where one previously did not exist.2.Streamlined