Single cell injection using nano pipette via nanorobotic manipulation system inside E-SEM

In this work, a nano injection system for single cell with small size (diameter: ~5 ¿m) is proposed. An environmental scanning electron microscope (E-SEM) nanorobotic manipulation system has been constructed with three units and seven degrees of freedom (DOFs) in total. It can provide a real-time observation with nanoscale resolution during the experiment. Micro pulling and FIB etching are used to control the tip size of nano pipette accurately. In situ penetration of single wild-type yeast cell (W303) is performed using nano pipette inside the E-SEM based on the nanorobotic manipulation system. Spouting experiment of Rhodamine 6G fluorescent solutions at femtoliter level from nano pipette is also achieved under the E-SEM environment. The results show that it is possible to perform small single cell injection using nano pipettes inside the E-SEM.