Microfluidic Surgery in Single Cells and Multicellular Systems

Abstract

Microscale surgery on single cells and small organisms have enabled major advances in fundamental biology and in engineering biological systems. Examples of applications range from wound healing and regeneration studies to the generation of hybridoma to produce monoclonal antibodies. Even today, these surgical operations are often performed manually, but they are labor-intensive and lack reproducibility. Microfluidics has emerged as a powerful technology to control and manipulate cells and multicellular systems at the micro- and nanoscale with high precision. Here, we review the physical and chemical mechanisms of microscale surgery, and the corresponding design principles, applications, and implementations in microfluidic systems. We consider four types of surgical operations: 1) Sectioning, which splits a biological entity into multiple parts, 2) ablation, which destroys part of an entity, 3) biopsy, which extracts materials from within a living cell, and 4) fusion, which joins multiple entities into one. For each type of surgery, we summarize the motivating applications and the microfluidic devices developed. Throughout the review, we highlight existing challenges and opportunities. We hope that this review will inspire scientists and engineers to continue to explore and improve microfluidic surgical methods.

Determining protein polarization proteome-wide using physical dissection of individual Stentor coeruleus cells

“…We show that intracellular protein distributions can be mapped by physically dissecting a large cell into pieces and analyzing the fragments. For smaller cells, microfabricated cutting devices capable of working at the cellular level, [37][38][39] combined with increasing sensitivity for proteomic analysis of small samples such as a single mammalian cell, [40][41][42][43] should allow proteomic dissection, as demonstrated in this work, to become a general method for subcellular proteomics complementary to existing methods. 10,11,44 Conclusions A substantial fraction of all proteins is localized with respect to the global anterior-posterior body axis of the Stentor cell.…”

Proteomic Dissection of a Giant Cell

“…The unusually large size of Stentor enables this strategy to be implemented using manual dissection with a glass needle, but such an approach will not work with smaller cells. However, a variety of microfabricated cutting devices capable of working at the cellular level are now being developed (Blauch 2017; Koppaka 2021; K Zhang 2021). This growing ability to cut tiny pieces of cells and tissues, combined with increasing sensitivity for proteomic analysis of small samples such as a single mammalian cell (Dou 2019; Zhu 2019; Tsai 2020; Schoof 2021), means that proteomic dissection, as demonstrated in this work, can provide a general method for subcellular proteomics that is complementary to existing methods that based on fractionation or imaging.…”