Single-Cell Patterning Based on Immunocapture and a Surface Modified Substrate

Abstract

Micropatterning technology offers powerful methods for biological analyses at the molecular level, enabling the investigation of cell heterogeneities, as well as high throughput detection. We herein propose an approach for single-cell patterning. The substrate was prepared using micro fabrication and surface modification processes, and the patterning template was prepared using bovine serum albumin and streptavidin, which can be employed for the patterning of any biological molecules containing biotin. Subsequent to photolithography, etching, chemical vapor deposition (CVD), and polyethylene glycol (PEG) treatment, the optimized patterns were obtained with high accuracy, strong contrast, and good repeatability, thus providing good foundations for the subsequent single-cell patterning. The surface passivation method was proven effective, preventing unwanted binding of the antibodies and cells. Based on this streptavidin template, the specific binding between the biotinylated antibodies and the antigens expressed on the surface of the cells was enabled, and we successfully achieved single-cell patterning with a single-cell capture rate of 92%. This single-cell array offers an effective method in the investigation of cell heterogeneity and drug screening. Further, these methods can be used in the final step for the screening and enrichment of certain cells, such as circulating tumor cells.