Abstract 5566: Streamlining tissue sample preparation: A comparative study on the efficiency and reproducibility of the Omni-Stainer™ dewaxing and HIER protocol

Abstract

Abstract The evolution of life sciences research has entailed a shift toward understanding cellular functions within tissue microenvironments, with an emphasis on cell- and tissue-specific relationships. A key challenge in these endeavors is the need for accurate data acquisition, which can be impeded by variability in the multistep manual assay preparations. In the quest for developing reliable and efficient tissue sample preparation workflows, two protocols using multiple dewaxing agents have been compared: the Omni-Stainer™ Dewaxing and HIER antigen retrieval protocol from Parhelia Biosciences, Inc. and the analogous one on Leica BOND Rx. The objective of this research was to assess the efficiency and reproducibility of these methods in the context of a four-color morphology immunostaining used for GeoMx DSP RNA and protein detection assays (Nanostring). We examined the efficiency of automated dewaxing using various non-toxic xylene alternatives, including C12-15 alkyl Benzoate, diethylene glycol, methyl octanoate or methyl decanoate, and compared them to the proprietary Leica Bond Dewaxing solution. Then, we compared the quality of post-HIER immunostaining using two different four-antibody panels on human jejunum samples. After several rounds of optimization, we found that the V4 of the open-source Omni-Stainer™ dewax-HIER protocol resulted in identical quality of morphology immunostaining compared to proprietary Leica Bond protocol. This study validates the Omni-Stainer™ Dewaxing and HIER protocol as a viable and efficient approach for automation of sample prep for GeoMx DSP studies. Citation Format: Nikolay Samusik, Yury Goltsev, Noemi Kedei. Streamlining tissue sample preparation: A comparative study on the efficiency and reproducibility of the Omni-Stainer™ dewaxing and HIER protocol [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 5566.