Hypoxia Methods and Protocols

This volume explores the latest ways to detect hypoxia in the context of cancer, including techniques to study gene expression changes, protein responses, and cellular adaptations to low-oxygen conditions. This book also covers new protocols to characterize hypoxia in tumors with high spatial resolution and novel therapeutic approaches that consider the complex microenvironments of solid tumors. 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.

Comprehensive and cutting-edge, Hypoxia: Methods and Protocols is a valuable tool to help cancer researchers and other scientists detect hypoxia in cancer and other disease pathologies.

Genetically Encoded Reporters to Monitor Hypoxia.- Detection of Hypoxia in 2D and 3D Cell Culture Systems using Genetically Encoded Fluorescent Hypoxia Sensors.- Mapping the Fate of Hypoxic Cells using an Irreversible Fluorescent Switch.- Application of a Specific and Sensitive NQO1 Turn-On Near-Infrared Fluorescence Probe for Live Cancer Cell and Xenografted Tumor Imaging in Nude Mice.- A Luciferase Reporter Assay to Detect Cellular Hypoxia In Vitro.- Oxygen Assessment in Tumors In Vivo using Phosphorescence Lifetime Imaging Microscopy.- Radionuclide Reporter Imaging to Visualize Tumor Hypoxia Ex Vivo and In Vivo.- Measuring Pericellular Oxygen Tension for In Vitro Cell Culture.- A Review of Hypoxia Imaging Using 18F-Fluoromisonidazole Positron Emission Tomography.- Photoacoustic Lifetime Imaging of Hypoxia.- Staining Hypoxic Areas in Frozen and FFPE Tissue Sections with Hypoxyprobe™.- Multiplex Immunofluorescence Staining Protocol for Dual Imaging of Hypoxia-Inducible Factors 1 and 2 on Formalin-Fixed Paraffin-Embedded Samples.- Detecting Hypoxia-Inducible Factor Levels and Activity.- Metabolomic Investigations into Hypoxia-Mediated Metabolic Reprogramming of Pancreatic Cancer Cells.- Measurement of Metabolic Alteration in Immune Cells under Hypoxia.- Evaluation of Oxygen Consumption Rates In Situ.- Cleanroom-Free Microfluidic Device for Natural Induction of Hypoxia in 2-D and 3-D Tumor Models.- Oxomer- and Reporter Gene-Based Analysis of FIH Activity in Cells.