Analysis of target mRNAs in the fixed-frozen human brain using a modified BaseScope-ISH Assay protocol
M. Carla Carisì, Owain W. Howell, Alwena H. Morgan and 1 more
STAR Protocols · 2022
doi:10.1016/j.xpro.2022.101896 (opens in a new tab)
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Abstract
Summary: We describe a modified BaseScope™ Assay protocol (ACDBio) for RNA in situ hybridization on fixed-frozen human brain tissue. The original protocol caused tissue detachment due to harsh tissue pre-treatment. We therefore optimized it to improve tissue stability while providing high stain quality in fragile post-mortem tissue from aged donors with advanced neurodegeneration. The main changes include two additional fixation steps and modifications to the pre-treatment protocol. We also describe tissue imaging and stain quantification using the open-source QuPath software.For complete details on the use and execution of this protocol, please refer to Hornsby et al. (2020).: Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Abstract licence: CC BY-NC
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