| Abstract [eng] |
Biological samples are intrinsically heterogeneous. Unraveling this heterogeneity is important for a better understanding of the biological functions of individual cells and their differences. Single-cell profiling often requires complex multi-step molecular biology techniques to isolate single cells and process their genetic material. However, despite recent progress in the single-cell analysis field, the high-throughput analysis remains limited due to numerous technological constraints. This doctoral thesis introduces a practical and innovative solution to enable a high-throughput and complex single-cell analysis. This achievement has been made possible by the development of semi-permeable microcapsules. The microcapsules are picoliter-nanoliter volume droplets enveloped by a thin hydrogel shell that acts as a semi-permeable membrane. Isolated single cells and high molecular weight biomolecules (e.g., gDNA, mRNA) are retained, while smaller molecules (e.g., enzymes, oligonucleotides, reagents) transverse the hydrogel shell by diffusion. Due to this semi-permeability, various enzymatic reactions and assays can be performed on a massive scale by simply transferring the microcapsules from one reaction tube to another. The microcapsules have been applied for single bacterial genome amplification, multiplex RT-PCR, RNA sequencing, and cell culture. The technology and results revealed in this thesis are likely to create a foundation for further development of high-throughput single-cell -omics methods. |
