Limiting Dilution
Capture and isolation of single cells are prerequisites for understanding these cellular variations. However, efficient single cell capture and isolation are complex tasks for any cell population. Therefore, many technologies for single cell capture and isolation have been developed.
Limiting Dilution
Limiting dilution is a technique for determining the frequency of a particular cell type in a complex cell mixture. According to the statistical distribution, the cell suspension is diluted to the point where no cell per aliquot is as low as a single cell per aliquot. The phenomenon of limiting dilutions has been used for decades to produce monoclonal cell cultures, for antibody production through hybridomas, and for single-cell-based assays.
Fig.1 Diagram of limiting dilution. (Hwang, 2018)
Using Pipetting Robot
Limiting dilution is one of the most well-liked, useful, and economical methods for single cell separation. Using pipettes or pipetting robots, this technology, which is based on the Poisson distribution, permits the quick and easy separation of individual cells from diluted cell solutions. To obtain an acceptable number of individual cells, the cell suspension is drastically diluted to a density of fewer than one cell per aliquot.
Fig.2 Diagram of a pipetting robot module. (Eggert, 2020)
Advantages of Limiting Dilution in Single Cell Isolation
- Simple, cost-effective, and reproducible.
- Viable Cell.
- High automation degree.
Limitations of Limiting Dilution in Single Cell Isolation
Although the technique is simple, it is not as efficient as the probability of obtaining a single-cell based on the Poisson distribution. One-third of cell culture plate wells actually contain single cells due to the statistical nature of the separation method.
- Lack of control over an individual cell.
- Require downstream process to prove the presence of single cells in a specific well.
Creative Biolabs offers efficient and detailed support to bring automation to your application workflows. Our Single Cell Sorting and Isolation Platform can provide highly customized comprehensive solutions with the best quality to advance our global clients' projects. Any requirements, please feel free to contact us for further communication about your project.
References
- Hwang, B.J.; et al. Single-cell RNA sequencing technologies and bioinformatics pipelines. Experimental & Molecular Medicine. 2018, 50: 1-14.
- Eggert, S.; et al. OpenWorkstation: a modular open-source technology for automated in vitro workflows. HardwareX. 2020, 8: E00152.
