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Cross-Membrane Penetration to Nucleus of Adherent Cells Using Micropipettes Made by Borosilicate Glass and Quartz

In the biomicromanipulation field, single-cell manipulation is receiving attention due to its potential applications in many aspects of biomedicine. Cell injection, as a typical cell micromanipulation technique, is devoted to introducing foreign substances into living cells by penetrating a fine micropipette into cell membranes. Direct access to the intracellular environment, especially the nucleus, provides an ideal way to gain insight into the dynamic processes of living cells. This paper presents the comparison of controlled penetrations of two types of adherent cells and their nuclei by two types of micropipettes, made of borosilicate glass and quartz respectively. Due to the differences in the cell properties (e.g., height, adhesion strength), two corresponding penetration strategies optimizing the puncture angle and puncture height, are proposed to improve the success rate of the puncture process and the survival rate of cells after the puncture. Real nucleus puncture experiments adopting the optimal puncture strategy for each type of cells and carried out by a semi-automatic system, indicated a better puncture effect by quartz pipettes. Through this method, functional materials such as peptides and nanoparticles can be potentially introduced to perturb the cell activities and then realize its full automation.