Single-cell protein analysis with NGS

Summary

Next-generation sequencing (NGS) has revolutionized research, enabling breakthroughs by profiling genomes and transcriptomes globally. Recent advancements allow single-cell sequencing, revealing cellular diversity and protein expression. By using barcoded antibodies, proteins can be analyzed as effectively as with flow cytometry, with the potential for high multiplexing. This approach facilitates multi-modal analysis, such as simultaneous gene and protein expression studies, and complements existing methods like flow cytometry. Illumina provides sequencing solutions across all stages of the process, empowering researchers with trusted technology for deeper biological insights, regardless of the scale or complexity of their samples.

Takeaways

• 😀 NGS (Next-Generation Sequencing) has revolutionized research, providing unbiased, global profiling of genomes and transcriptomes for over 10 years.
• 😀 Advances in NGS have expanded our understanding of biological systems in both health and disease.
• 😀 Single-cell sequencing has emerged, allowing for in-depth analysis of individual cells and revealing cell diversity and heterogeneity.
• 😀 Single-cell sequencing has contributed to major insights, such as understanding the specialized cells in the immune system.
• 😀 Protein analysis is now possible through single-cell sequencing, using barcoded oligonucleotides instead of traditional fluorophores in flow cytometry.
• 😀 Barcodes used in protein analysis have vast multiplexing potential, with over 200 proteins able to be profiled simultaneously.
• 😀 Single-cell sequencing enables multi-modal analysis, where multiple biological molecules (like genes and proteins) can be studied in the same cell.
• 😀 Single-cell sequencing complements flow cytometry by providing unbiased RNA or chromatin accessibility data, followed by more specific cell counting.
• 😀 Single-cell sequencing can be used to validate discoveries made with flow cytometry and enhance understanding.
• 😀 Commercial solutions and academic analysis packages are available to support every step of single-cell sequencing, from tissue prep to analysis.
• 😀 Illumina sequencing technology is proven and trusted, with over 200,000 publications demonstrating its effectiveness across various research fields.

Q & A

What is the main advancement enabled by Next Generation Sequencing (NGS) over the past 10 years?

NGS has enabled researchers to profile the genome and transcriptome in a global and unbiased manner, leading to groundbreaking findings that have expanded our understanding of biological systems in both health and disease.

How has single-cell sequencing improved the study of biological systems?

Single-cell sequencing allows for the study of biological systems at the level of individual cells, revealing cell diversity and heterogeneity across different research fields. This has provided new insights into complex systems like the immune system.

What is the key difference between single-cell protein analysis and traditional flow cytometry?

In single-cell protein analysis, antibodies are labelled with barcoded oligonucleotides instead of fluorophores, which enables more accurate protein abundance data to be read by Illumina sequencers, providing data as robust as that generated by flow cytometry.

How does the barcode space in single-cell sequencing enhance its potential?

The barcode space in single-cell sequencing is virtually infinite, allowing for high levels of multiplexing. This makes it possible to profile over 200 proteins simultaneously, providing far more information than traditional methods.

What benefits does multi-modal analysis in single-cell sequencing provide?

Multi-modal analysis allows researchers to study multiple biological molecules in the same cell, such as simultaneously examining gene and protein expression. This deeper analysis helps to uncover the relationship between these two classes of molecules.

How can single-cell sequencing complement flow cytometry?

Single-cell sequencing can complement flow cytometry by offering an unbiased analysis of RNA or chromatin accessibility, which can then be followed up with cell counting for further investigation. It also helps to validate discoveries made using flow cytometry.

What role does cell sorting play in single-cell sequencing?

Cell sorting can be used to enrich specific cells or cell types, allowing for deeper and more focused single-cell sequencing. This ensures more detailed insights into particular cell populations.

What are the key stages in the single-cell sequencing workflow?

The single-cell sequencing workflow consists of tissue preparation, single-cell isolation and library preparation, sequencing, and analysis. Commercial solutions are available for each of these steps to streamline the process.

How does Illumina contribute to single-cell sequencing research?

Illumina offers a range of sequencing systems, from small benchtop sequencers to larger production-scale instruments, providing trusted data for over 200,000 publications. They also offer commercial and open-source analysis packages to maximize biological insights.

What makes Illumina's technology a reliable choice for single-cell sequencing?

Illumina's technology is trusted by researchers worldwide, with over 200,000 publications using its systems. Their sequencing solutions are proven to provide accurate and reliable data, supporting a wide range of biological research needs.