Alternative cell counting and viability detection using NexGreen/PI fluorescent stain on multiple low- and high-throughput image cytometry platforms

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Presentation Time: 03:15 PM - 04:30 PM
Poster Board Number: B908
Abstract ID: 5222

Presenting Author:
Carolina Nitta , Consumables Development Manager at Revvity Hlth. Sci.

Abstract:

In the past decade, the increase in cell and gene therapy products such as biologics, antibodies, and CAR-T has significantly increased the need for precise, robust, and consistent cell counting and viability measurements. In this work, we demonstrate a novel fluorescence-based dye, NexGreen/PI, capable of live and dead cell detection comparable to AO/PI in multiple cell buffer/media conditions. Jurkat cells of different viabilities (low, intermediate, and high) were stained in parallel with either NexGreen/PI or AO/PI for 1-2 minutes and measured on multiple image cytometer platforms including low-throughput and high-throughput instruments. Both NexGreen/PI and AO/PI dyes are used as equal volumes added to cell samples. Additionally, we tested if difficult to analyze cells in “spent” media (typically with a lower pH) or in PBS can be stained using NexGreen/PI compared to AO/PI. Results on the high-throughput platforms show that NexGreen/PI staining can discriminate between live and dead Jurkats, analogous to AO/PI. Measured viability and cell concentration of all samples using NexGreen/PI was within 5% of the paired AO/PI-stained samples. Low-throughput instrument results are also similar. Our experiments demonstrate that NexGreen/PI is scalable and can be of significant value for use in automation and high-throughput systems where multiple samples in different mediums can be precisely measured.