Discover the Secrets of In Situ Hybridization in Electron Microscopy with This Cutting-Edge Guide

Chapter 1: Overview of ISH-EM

This chapter provides a detailed overview of the principles and applications of ISH-EM. It covers the basics of ISH, including probe design, hybridization methods, and signal detection. The chapter also discusses the integration of ISH with EM, highlighting the advantages and limitations of different approaches.

In Situ Hybridization in Electron Microscopy (Methods in Visualization)

by Lluís Montoliu

4 out of 5

Language	:	English
File size	:	35445 KB
Text-to-Speech	:	Enabled
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Enhanced typesetting	:	Enabled
Print length	:	472 pages

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Chapter 2: Probe Design and Labeling

Chapter 2 focuses on the crucial aspect of probe design and labeling. It explains the various types of probes used in ISH-EM, including DNA, RNA, and peptide nucleic acid (PNA) probes. The chapter also covers the methods for labeling probes with electron-dense tags, such as gold particles or horseradish peroxidase (HRP).

Chapter 3: Hybridization Methods

Chapter 3 delves into the different hybridization methods used in ISH-EM. It provides detailed protocols for optimizing hybridization conditions, including temperature, humidity, and buffer composition. The chapter also discusses techniques for enhancing signal-to-noise ratios, such as blocking non-specific binding and using amplification methods.

Chapter 4: Detection and Imaging

Chapter 4 describes the techniques for detecting and imaging the hybridization signals in ISH-EM. It covers both traditional methods, such as silver enhancement and immunogold labeling, and advanced approaches, such as correlative light and electron microscopy (CLEM). The chapter also discusses the use of computational tools for image analysis and data quantification.

Chapter 5: Applications in Cell and Tissue Biology

Chapter 5 showcases the diverse applications of ISH-EM in cell and tissue biology. It highlights studies on gene expression patterns in embryos, tumor cells, and stem cells. The chapter also provides insights into using ISH-EM to investigate the localization of viral and bacterial pathogens.

Chapter 6: Challenges and Future Directions

Chapter 6 discusses the challenges associated with ISH-EM, such as probe accessibility, background reduction, and resolution limitations. It also explores emerging technologies that may overcome these challenges and push the boundaries of ISH-EM in the future.

In Situ Hybridization In Electron Microscopy Methods In Visualization is an indispensable resource for researchers using ISH-EM. With its comprehensive coverage of the subject, from basic principles to advanced applications, this book provides a roadmap for harnessing the power of ISH-EM to unlock new insights into cellular and molecular biology.