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This detailed volume serves as a collection of methods for single-cell protein analysis, created by combining different protocols, taking advantage of new emerging technologies, and improving upon conventional methods to guide researchers aiming to perform protein analysis in single cells. Ranging from simple to complex, conventional to the most current technologies, these chapters offer readers the ability to choose the best suited methodologies for them, based on the sample type and the available technologies or equipment. Written for the highly successful Methods in Molecular Biology series, 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 practical, Single-Cell Protein Analysis: Methods and Protocols aims to help researchers utilizing single-cell protein analysis in their studies as well to inspire the development of the next generation of improved protein analysis methods in single cells.
List of contents
Immunochemistry Analysis Using Chromogenic Substrates on Tissue Sections.- Indirect Immunofluorescence of Tissue Sections.- Flow Cytometry for Beginners: Hints and Tips for Approaching the Very First Single-Cell Technique.- High-Dimensional Immunophenotyping with 37-Color Panel Using Full-Spectrum Cytometry.- Detection of Cytokine Secreting Cells by Enzyme-Linked Immunospot (ELISpot).- Detection and Enumeration of Cytokine Secreting Cells by FluoroSpot.- Single Cell Protein Profiling by Microdroplet Barcoding and Next Generation Sequencing.- Single Cell Proteomics Using Multiplexed Isobaric Labeling for Mass Spectrometric Analysis.- Customizing Maxpar Direct Immune Profiling Assay with Additional Surface Marker and Intracellular Cytokine Staining Workflows for Expanded Mass Cytometry Panels.- High-Dimensional Tissue Profiling by Multiplexed Ion Beam Imaging.- Ultra-Sensitive Quantification of Protein and mRNA in Single Mammalian Cells with Digital PLA.- High-Throughput Multimodal Single-Cell Targeted DNA and Surface Protein Analysis Using the Mission Bio Tapestri Platform.- Simultaneous Analysis of Single-Cell Transcriptomes and Cell Surface Protein Expression of Human Hematopoietic Stem Cells and Progenitors Using the 10x Genomics Platform.- Generation of Centered Log-Ratio Normalized Antibody-Derived Tag Counts from Large Single-Cell Sequencing Datasets.- Simultaneous Quantification of Single-Cell Proteomes and Transcriptomes in Integrated Fluidic Circuits.- Combined Measurement of RNA and Protein Expression on a Single-Cell Level.- Simultaneous DNA, RNA, and Protein Analysis from Single Cells Using a High-Throughput Microfluidic Workflow for Resolution of Genotype-to-Phenotype Modalities.
