Single-Cell Analysis
Single-cell analysis methods enable researchers to gain an in-depth understanding of cell programming and differentiation. However, conventional techniques are insufficient to capture the differences between cells and the heterogeneity of cultures. "Microfluidics" is related to handling and processing very small amounts of fluids and chemical and biological analytes, which makes it ideal for single-cell analysis.
The microfluidic technology used for single-cell analysis refers to the process of separating, manipulating, and analyzing cells with single-cell resolution in a microfluidic chip. Alfa Chemistry has developed various microfluidic devices to separate, manipulate and analyze single cells.
Alfa Chemistry Analysis Microfluidic Chips for Single Cell Help
With solid R&D experience in the field of microfluidics, we have helped customers successfully design and develop a series of microfluidic chips that has proven to be a powerful tool for single-cell analysis. Not only can it be used for single-cell isolation, but it can also provide researchers with new methods for genomics and transcriptomics research.
Microfluidic Method for Single-Cell Separation
Isolation is the first step for working with individual cells. It may be necessary to use microfluidic cell separation methods to first separate two different cell types from each other, and then separate specific cell types. Our microfluidic single-cell separation methods include, but are not limited to the following:
- Microwell array
The important parameters here are the size of the pores and the modification of the surface. The well allows the capture and release of individual cells by reversing the flow in the experiment. We have manufactured microchips with approximately 10000 holes.
- Droplets
Droplet-based microfluidics is the most popular single-cell analysis method. In droplet microfluidics, the flow conditions can be adjusted to encapsulate a single cell in each droplet, where cells can be lysed, DNA/RNA extraction, etc. The Poisson distribution is usually used to adjust the cell concentration to ensure that the droplet does not contain more than one cell.
We have developed a variety of microfluidic methods for the post-modification of droplets and the analysis of these cell-encapsulated droplets. Depending on the type of analysis, different reagents can be used in the droplet, and different protocols will be used for analysis.
- Traps
Microfluidic manufacturing allows the creation of active and passive capture methods. In the passive method, the common hydrodynamic trap is a U-shaped trap, in which a single cell is trapped in the U-shaped barrier, while the bypass microfluidic channel trap draws the cells into the side channel. For active methods, optical tweezers can use a highly focused laser beam to capture single cells and transfer them to the desired location.
Droplet Microfluidic Technology for Genome Research
Single-cell sequencing of the genome can better understand the genotypic basis of various phenotypes. Droplet microfluidic single-cell genomics aims to separate high-throughput single cells from picoliter to nanoliter droplets and amplify their DNA content. Depending on whether the target is the entire genome or part of the genome, different methods can be used for nucleic acid amplification.
For targeted genome analysis, we encapsulate cells, primers, and fluorescent probes in droplets of the hydrogel. The droplets are then subjected to PCR cycles, followed by gelation and washing. Here, the fluorescence intensity of the cross-linked gel will be used for analysis.
Droplet Microfluidic Technology for Transcriptomics Research
Single-cell transcriptomics aims to study the messenger RNA (mRNA) content of single cells. The two droplet-based microfluidic technologies we use are Drop-seq and inDrop. Both of them use droplet microcapsules to encapsulate single cells to analyze their mRNA content.
Advantages of Microfluidic Chips in Single-Cell Analysis
- The chip has a flat geometry, making it easier to observe and detect.
- The channel diameter of the microfluidic chip is compatible with the size of a single cell.
- Its network type of 2D or 3D channel is very easy to manipulate single-cell-scale targets.
- The chip has a smaller geometric size, and can obtain a larger electric field strength under a smaller voltage.
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