title: Intracellular tracking of single native molecules with electroporation-delivered quantum dots.
abstract: Quantum dots (QDs) have found a wide range of biological applications as fluorophores due to their extraordinary brightness and high photostability that are far superior to those of conventional organic dyes. These traits are particularly appealing for studying cell biology under a cellular autofluorescence background and with a long observation period. However, it remains the most important open challenge to target QDs at native intracellular molecules and organelles in live cells. Endocytosis-based delivery methods lead to QDs encapsulated in vesicles that have their surface biorecognition element hidden from the intracellular environment. The probing of native molecules using QDs has been seriously hindered by the lack of consistent approaches for delivery of QDs with exposed surface groups. In this study, we demonstrate that electroporation (i.e., the application of short electric pulses for cell permeabilization) generates reproducible results for delivering QDs into cells. We show evidence that electroporation-based delivery does not involve endocytosis or vesicle encapsulation of QDs. The amount of QD loading and the resulting cell viability can be adjusted by varying the parameters associated with the electroporation operation. To demonstrate the application of our approach for intracellular targeting, we study single-molecule motility of kinesin in live cells by labeling native kinesins using electroporation-delivered QDs. We envision that electroporation may serve as a simple and universal tool for delivering QDs into cells to label and probe native molecules and organelles.
authors.affiliation: School of Biomedical Engineering and Sciences, Virginia Tech-Wake Forest University , Blacksburg, Virginia 24061, United States.
authors.forename: Chen
authors.initials: C
authors.lastname: Sun
authors.forename: Zhenning
authors.initials: Z
authors.lastname: Cao
authors.forename: Min
authors.initials: M
authors.lastname: Wu
authors.forename: Chang
authors.initials: C
authors.lastname: Lu
mesh_terms: D000818:Animals; D016466:CHO Cells; D002470:Cell Survival; D002478:Cells, Cultured; D003412:Cricetulus; D018274:Electroporation; D042541:Intracellular Space / Q000378:metabolism*; D016547:Kinesins / Q000032:analysis*; D046210:Microfluidic Analytical Techniques; D045663:Quantum Dots
chemical_list: D016547:Kinesins