Single Molecule Counting Of Biomolecule Of Single Cell Based On The Self-Priming Compartmentalization Digital PCR Chip

As the basic unit of organism structure and functions, cell occupies an important position in biomedical research. Analysis for genome and transcriptome of cell has profound and lasting significance in the field of differentiation, development and the origins of many serious diseases. Currently, traditional cytogenetic analysis is still confined to get average results of analyzing a large number of cells, which ignores the heterogeneity between cells and the possibility that the characteristics of rare cells can be masked by the majority. Thus, genetic analysis at single-cell level needs to be further studied. In this paper, novels for single cell genetic analysis and single cell protein analysis are reviewed. Novels for single cell analysis based on microfluidic platform are comprehensively summarized, classified and described. Based on this, the paper applied an integrated self-priming compartmentalization chip to precise quantifications of copy number of 18s, β-actin and PLAU within individual A459 cells, optimized single-cell digital polymerase chain reaction (dPCR) reaction system and reaction conditions in nanoliter reactions, and expanded quantifications to the single cell protein molecules by digital PLA. This paper has two innovations. One is that by using digital PCR microfluidic chip this paper accomplishes the quantifications of copy number for three different abundance of genes within individual cells, which proves the heterogeneity between cells. The other one is through digital proximity ligation assay (PLA) based on microfluidic chip this paper expands the application of microfluidic chip from nucleic acid quantifications to that of protein within single cells. These results may provides an important tool for the study of the origin and early diagnosis of diseases, prenatal diagnosis, cell differentiation and development. Further research can establish a link between quantity of nucleic acid molecules and levels of corresponding protein content within a single cell and offer a new novel for single cell genome, transcriptome and proteome research.