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A cell is a basic unit of biological structure and function, varying broadly in type and state. The characterization of single-cell identity and function will accelerate biological discovery and development, as the understanding of functional abilities and responses of each cell type. As a first-class provider in the microfluidic chip market, Creative Biolabs has presented a reliable technology platform to support single-cell sequencing services and to analyze a large number of individual cells simultaneously. Aided by our sophisticated engineers, a number of sequencing strategies, such as based on droplet microfluidics, have been introduced to characterize complex tissues with varieties of cell lines and meet the demand of rapid, scalable approaches.

Single Cell Sequencing

Next-Generation sequencing (NGS) is a technology enabling the sequencing of the entire human genome within one day. Firstly, the entire genome is divided into millions of fragments and sequenced in parallel. Next, data collected from individual fragments are mapped to a reference human genome through bioinformatics analysis. The process is repeated many times to obtain accurate information on any DNA mutation. As well, NGS can be used for extracting data from desired genetic locations and has better sensitivity towards low-frequency variants because of its high sequencing depth.

NGS along with microfluidic technique has multiple applications in DNA sequencing, RNA sequencing, and ATAC sequencing (chromatin) of many kinds of individual cells. Some of the major technologies have been developed recently, especially single-cell RNA-Seq that displays quantitatively comparable to multiplexed quantitative PCR (qPCR).

Fig. 1 Microfluidic chip for single cell analysis.¹

Microfluidic Platform & Service at Creative Biolabs

The ability to associate single-cell genetic information to cellular phenotypes could present a detailed insight into human physiology and disease mechanisms that is hard to infer from bulk cell analysis. Microfluidic technologies are valuable and attractive for single-cell manipulation owing to strict handling and low risk of contamination. Incorporating microfluidic platforms into research and clinical workflows will fit an unmet need in biology, depicting the highly precise and informative results necessary to develop novel therapies and monitor patient outcomes. At Creative Biolabs, we’d like to introduce the following strategies to perform single-cell sequencing services in terms of microfluidic technologies.

Fig.2 Single-cell transcriptome sequencing sensitivity. (Street, 2014)

• Microfluidic droplet-based single-cell sequencing (Drop-seq)

In droplet-based sequencing, individual cells are separated in nanolitre-scale aqueous compartments that work as tiny reaction rooms for PCR and reverse transcription. This method can analyze thousands of cells in parallel for mRNA transcripts while keeping a record of the transcript’s cell of origin via genomic barcodes. The barcodes offer a molecular memory, serving as an identifier-read at sequencing to decode specific information of every single cell. Here, we have two types of beads used to synthesize primers, including simple microparticles and hydrogel microparticles. After preparing the single-cell suspension and microparticles, the individual cell can be encapsulated with microparticles in droplets, using a custom-designed microfluidic chip.

• Microfluidic diffusion-based single-cell RNA sequencing (scRNA-seq)

Our microfluidic devices provide a high-efficacy and low-input platform for single-cell RNA sequencing. Today, we have developed microfluidic diffusion-based RNA sequencing for supervising scRNA-seq with a diffusion-based reagent swapping scheme. This device merges cell capturing, cell lysis, reverse transcription, and PCR amplification all into one simple microfluidic chip. This sequencing system creates high data quality that is comparable to current scRNA-seq methods while executing a simple device design that allows multiplexing. The robustness and scalability of microfluidic diffusion-based scRNA-seq chips will be significant for transcriptomic research of scarce cell samples.

• Microfluidic single-cell whole-transcriptome sequencing

The low abundance of RNA in a single cell hinders efficacious, consistent reverse transcription and amplification of cDNA, blocking accuracy and obscuring biological variability with complex technical noise. Notably, our experts have designed a microfluidic technique to prepare cDNA from the single cells for high-throughput transcriptome sequencing. The microfluidic chip device promotes single-cell manipulation, minimizes contamination, and moreover, displays improved detection sensitivity and measurement precision, which is essential for differentiating biological variation from high technical noise.

Key-benefits

Fig.3 Advantages of microfluidics for single-cell genetic analysis. (Thompson, 2014)

• Multiple single-cell sequencing strategies and customized microfluidic chips can be chosen to satisfy clients’ special needs;
• Our microfluidic single-cell sequencing techniques allow for high-throughput and detailed genetic analyses;
• Increased accuracy, decreased reagent cost, and minimized consumption of rare or expensive samples;
• Skillful technicians and optimized protocols result in high efficacy and a short period of time.

High-throughput, microfluidic-based single-cell sequencing is rapidly stepping forward with technical upgrades and additional software to extend the scientific understanding of genomics studies at the level of single cell and single nucleus. As a well-recognized expert in the microfluidic field, Creative Biolabs has developed a series of advanced platforms and personalized strategies to accomplish single-cell sequencing services. We always pay attention to the current and future potential utilization of microfluidics at all stages of single-cell genetic analysis, including cell enrichment and capture, single-cell compartmentalization, and manipulation, as well as detection and sequencing analyses. If you’ like to know more about our custom microfluidic chips, please directly contact us or send us an e-mail with your requests.

References

1. Streets, Zhang, et al. " Microfluidic single-cell whole-transcriptome sequencing " Biomolecules 111.19 (2014): 7048-7053.
2. Thompson, Paguirigan, et al. " Microfluidics for single-cell genetic analysis. " Lab Chips 14.17 (2014): 3135-3142.

For Research Use Only. Not For Clinical Use.