This fifth edition volume expands on the previous editions with updated discussions on the many new in situ hybridization (ISH) techniques used by researchers today. New developments in probe designs, detection systems, specificity and sensitivity improvements, and multiplexing combinations are explored. Chapters in this book are organized into seven sections and cover general applications; methods for DNA ISH; methods for cultured cells; methods for wholemount and plant material; automated methods for RNA; multiplexing and combined methods; and targeted selective methods and single molecule detections. 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. Cutting-edge and thorough, In Situ Hybridization Protocols, Fifth Edition is a valuable resource for both novice and expert scientists interested in learning more about this exciting and advancing field.