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Full Description
This sixth edition volume expands on the previous editions with a discussion of the latest methods used to detect RNA or DNA species in situ, including both short and long RNAs, and coding and non-coding RNAs. The chapters in this book are organized into five parts: RNA detection techniques; RNA and protein co-detection; 3D and wholemount ISH; spatial transcriptomics approaches; and image analysis and quantification. Some of the topics covered in this volume are detection of viral pathogens using RNAscope™; detection of therapeutic antisense oligonucleotides using miRNAscope assays; RNAscope HiPlex in tissue sections and wholemounts; spatial transcriptomics platforms with accompanying data processing; and various image analysis platforms for downstream quantitative assessment. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and thorough, In Situ Hybridization Protocols, Sixth Edition brings together a collection of protocols, along with practical notes and recommendations, and offers researchers a comprehensive introduction to the current leading edge of in situ hybridization.
Contents
Automated In Situ Hybridization for Detection of Viral Pathogens using RNAscope™.- Use of Automated BaseScope Assay on the Leica Bond RX.- Spatial Analysis of Perivascular Adipose Tissue Gene Expression with SABER-FISH.- HiPlex RNAscope™ for In Situ Analysis of Gene Expression during Murine Thymocyte Differentiation.- Detection of Unlabelled Oligonucleotides in FFPE Tissue Sections using Automated miRNAscope Assays.- High-Sensitivity In Situ Hybridization DNAscope for Detection of Papillomavirus or Epstein-Barr Virus Genomes Combined with Immunofluorescent Staining in Adherent Cells.- Assessing Cell-Specific Delivery and Efficacy of Therapeutic Oligonucleotides using Combined RNAscope Plus and Protein Multiplexing in FFPE Tissue.- Protease-Free RNAscope and Sequential Immunofluorescence Methods for Manual Co-Detection of RNAs and Proteins.- Multiomic Detection of RNA, Protein, and Protein-Protein Proximity at Subcellular Resolution using Automated Protease-Free RNAscope™ Workflows.- Multiplexed Fluorescent In Situ Hybridization in Plant-Parasitic Nematodes.- 3D RNA Detection in Millimeter-Scale Tissue using RNAscope HiPlex.- LNA-Probe-Based MicroRNA In Situ Hybridization in Brain Organoids.- RNAscope™ In Situ Hybridization for the Detection of a Long Non-Coding RNA in an Organ-On-A-Chip.- GeoMx Spatial Transcriptomics for Comprehensive Glioblastoma Molecular Profiling.- Spatial Single Cell RNA Profiling of Human Liver FFPE Tissue Microarrays using Xenium In Situ.- Plant-Compatible Xenium In Situ Sequencing: Protocol for Spatial Transcriptomics in Medicago truncatula Roots and Nodules.- Improving Spatial Transcriptomics with Dual-Color Coding and Cell Painting.- QuPath for RNAscope Image Analysis.- Image Analysis Approaches for In Situ Hybridization Technologies.- Overcoming Variability in RNAscope Image Analysis: A HALO-Based Approach.
