Main description:

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.

Contents:

1. Immunochemistry Analysis Using Chromogenic Substrates on Tissue Sections

Preethi Vijayaraj

2. Indirect Immunofluorescence of Tissue Sections

Cody J. Aros

3. Flow Cytometry for Beginners: Hints and Tips for Approaching the Very First Single-Cell Technique

Valentina Proserpio, Laura Conti, and Salvatore Oliviero

4. High-Dimensional Immunophenotyping with 37-Color Panel Using Full-Spectrum Cytometry

Marco A. Fernandez, Hammad Alzayat, Maria C. Jaimes, Yacine Kharraz, Gerard Requena, and Pedro Mendez

5. Detection of Cytokine Secreting Cells by Enzyme-Linked Immunospot (ELISpot)

Bernt Axelsson

6. Detection and Enumeration of Cytokine Secreting Cells by FluoroSpot

Bernt Axelsson

7. Single Cell Protein Profiling by Microdroplet Barcoding and Next Generation Sequencing

Samuel C. Kim, John R. Haliburton, Zev J. Gartner, and Adam R. Abate

8. Single Cell Proteomics Using Multiplexed Isobaric Labeling for Mass Spectrometric Analysis

Akos Vegvari, Jimmy E. Rodrigues, and Roman A. Zubarev

9. Customizing Maxpar Direct Immune Profiling Assay with Additional Surface Marker and Intracellular Cytokine Staining Workflows for Expanded Mass Cytometry Panels

Carlene Petes, Stephen K.H. Li, Shariq Mujib, Michelle M. Poulin, Noah Saederup, Andrew A. Quong, and Christina Loh

10. High-Dimensional Tissue Profiling by Multiplexed Ion Beam Imaging

Ofer Elhanani, Leeat Keren, and Michael Angelo

11. Ultra-Sensitive Quantification of Protein and mRNA in Single Mammalian Cells with Digital PLA

Jing Lin and Savas Tay

12. High-Throughput Multimodal Single-Cell Targeted DNA and Surface Protein Analysis Using the Mission Bio Tapestri Platform

David W. Ruff, Dalia Dhingra, Kathryn Thompson, Jacqueline A. Marin, and Aik T. Ooi

13. Simultaneous Analysis of Single-Cell Transcriptomes and Cell Surface Protein Expression of Human Hematopoietic Stem Cells and Progenitors Using the 10x Genomics Platform

Nicole Mende, Elisa Laurenti, Berthold Gottgens, and Nicola K. Wilson

14. Generation of Centered Log-Ratio Normalized Antibody-Derived Tag Counts from Large Single-Cell Sequencing Datasets

Benjamin Lacar

15. Simultaneous Quantification of Single-Cell Proteomes and Transcriptomes in Integrated Fluidic Circuits

Mandi Wong, Carol Kosman, Liane Takahashi, and Naveen Ramalingam

16. Combined Measurement of RNA and Protein Expression on a Single-Cell Level

Valentina Russo, Nadia Brasu, and Luigia Pace

17. Simultaneous DNA, RNA, and Protein Analysis from Single Cells Using a High-Throughput Microfluidic Workflow for Resolution of Genotype-to-Phenotype Modalities

Dalia Dhingra, Aik T. Ooi, and David W. Ruff