Abstract

Abstract In this study, we introduce an innovative microwell-based approach, offering the capability to capture up to 500,000 single cells and show stability of a partially used 8-lane microwell cartridge up to 6 months after first use. Stability of microwell cartridges used for single-cell capture and the integrity of the individual lanes are crucial factors in genomics research to ensure the accuracy and reliability of generated data. Stability is paramount to ensure consistent results over multiple experiments. High-quality materials and manufacturing processes are essential to prevent wear and tear, ensuring that the microwell cartridges function optimally for an extended period. Integrity between the individual lanes must also be intact to prevent cross-contamination between samples. To demonstrate partial use stability of the 8-lane microwell cartridge, multiple assay combinations were performed on different lanes of the cartridge, collected at different time points. Whole transcriptome amplification (WTA) together with AbSeq antibody-oligonucleotide conjugates for cellular indexing of transcriptomes and epitopes (CITE-Seq) was performed on peripheral blood mononuclear cells (PBMCs), loaded and captured in Lanes 1 and 2, on Day 1; sample multiplexing was used with targeted mRNA assay on Jurkat and Ramos cells, loaded and captured in Lanes 3 and 4, after four months; and WTA together with the AbSeq Assay was performed on PBMCs, loaded and captured in Lanes 5 and 6, after six months. The different time points collected were compared to a single-lane microwell cartridge. Our data showed that assay performance was not compromised with the storage of a partially used cartridge, and there was no contamination detected between the lanes. High-throughput cell load and capture using WTA and the AbSeq Assay also showed that each lane of the 8-lane microwell cartridge was able to process up to 65,000 cells with no limit of detection on assay sensitivity. By maintaining cartridge stability and stringent measures to prevent contamination, researchers can ensure the reliability and accuracy of their data. This is vital for various applications such as developing targeted immunotherapies and precision medicine approaches, ultimately enhancing our capacity to combat a wide range of diseases. For Research Use Only. Not for use in diagnostic or therapeutic procedures.BD and BD Rhapsody are trademarks of Becton, Dickinson and Company or its affiliates. © 2023 BD. All rights reserved. Citation Format: Hye-Won Song, Ricelle A. Acob, Zorine Hlathu, Xueying Zhao, Jamie Moskwa, Larry Wang, Aruna Ayer. Stability and performance of a partially used 8-lane microwell cartridge on single-cell capture and immune response profiling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 333.

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