Generation of stable monoclonal cell lines is required for a wide variety of applications, including the development of biologics, such as therapeutic antibodies, or to create CRISPR engineered cells for disease in a dish models While the demand for engineered cell lines has increased, production of monoclonal lines is still limited by the current technology for single cell sorting and dispensing. Here we contrast the limitations of traditional technologies with newer technologies, such as microfluidic sorters and dispensers (WOLF ® Cell Sorter, NanoCellect ® Inc that can analyze, sort and dispense single cells into 96 or 384 well plates with high viability and outgrowth We highlight that both robust immortalized cell lines (e g CHO) and more sensitive cells can be used to generate monoclonal cell lines in a sterile format with little cellular stress The results reveal that the WOLF cell sorting platform can efficiently isolate targeted single cells while maintaining high viability and can be used for the isolation of single cell clones for applications such as CRISPR modified stem cells and antibody production CHO cells.