Recent research advance in single cell sequencing for rare cell samples
|22.2.2020||Posted by tactical33 under Advertising & Marketing|
Single-cell sequencing technology is a cutting-edge technology for life sciences that has developed rapidly in recent years. It reveals cell’s gene structure and gene expression status at a single-cell level, reflects the heterogeneity between cells, and provides a unique perspective for life science research. In recent years, single-cell sequencing technology has played an important role in tumors, developmental biology, microbiology, and neuroscience, and has become a hot research tool with great potential in life sciences.
Conventional commercial platforms such as 10X Genomics, BD Rhapsody and other systems are mainly suitable for the isolation and library construction of large-scale cells, but for extremely rare samples, such as circulating tumor cells, embryonic stem cells, etc., are still in great challenges for capture and sequencing. Traditional techniques are difficult to avoid the loss of rare samples, and it is difficult to optimize the quality of library construction at the single-cell level to meet the needs of deep sequencing.
Recently, a research has obtained new progress in single-cell sequencing for rare cell samples.
The research aimed at the analysis of rare cell samples, proposed a novel method to improve the efficiency of single-cell transcriptome sequencing, and developed a microfluidic chip integrated with dozens of cell capture-manipulation units. In the microfluidic chip, the micro-sieve structure combined with the hydrodynamic design is integrated with the single-cell manipulation unit, which can achieve a single-cell capture rate close to 100% and minimize the loss of cells, so this unique chip design is ideal for rare biological samples. A new liquid-gel phase transition strategy is introduced into the microfluidic chip, which can quickly encapsulate single cells into skin-upgraded hydrogel droplets, making it easy to manipulate and maintain cell viability, helping to achieve subsequent single-cell transcriptome amplification reactions. At the same time, single-cell transcriptome amplification is performed in a micro-reaction system for skin upgrade, which can improve the reverse transcription efficiency in RNA-Seq. With this novel microfluidic platform, researchers can capture dozens of human embryonic stem cells (hESCs) at the same time, and then manipulate them with precise microvalves in the chip to obtain dozens of picoliters of single-cell samples. High-quality libraries for deep single-cell transcriptome sequencing. The single-cell transcriptome sequencing data in this study verified that the method can improve the efficiency of transcriptome amplification and obtain high-quality cDNA libraries for sequencing analysis. Sequencing results confirmed that the H9 community of human embryonic stem cells was heterogeneous and had a broad spectrum of pluripotency.
This research builds a low-cost, stable and reliable microfluidic platform that can achieve efficient single-cell sequencing preprocessing, which is a clinical application for single-cell sequencing, especially for rare samples. The accurate analysis provides a new way of thinking, which helps scientists overcome the key technical difficulties in current liquid biopsy research.