Using Fluid Walls for Single-Cell Cloning Provides Assurance in Monoclonality

Single-cell isolation and cloning are essential steps in many applications, ranging from the production of biotherapeutics to stem cell therapy. Having confidence in monoclonality in such applications is essential from both research and commercial perspectives, for example, to ensure that data are of high quality and regulatory requirements are met. Consequently, several approaches have been developed to improve confidence in monoclonality. However, ensuring monoclonality using standard well plate formats remains challenging, primarily due to edge effects; the solid wall around a well can prevent a clear view of how many cells might be in a well. We describe a method that eliminates such edge effects: solid confining walls are replaced by transparent fluid ones, and standard low-cost optics can confirm monoclonality.