Microinjection into single cells in brain tissue is a powerful technique to study and manipulate neural stem cells. However, such microinjection requires expertise and is a low-throughput process. We developed the “Autoinjector”, a robot that utilizes images from a microscope to guide a microinjection needle into tissue to deliver femtoliter volumes of liquids into single cells. The Autoinjector enables microinjection of hundreds of cells within a single organotypic slice, resulting in an overall yield that is an order of magnitude greater than manual microinjection. The Autoinjector successfully targets both apical progenitors (APs) and newborn neurons in the embryonic mouse and human fetal telencephalon. We used the Autoinjector to systematically study gap-junctional communication between neural progenitors in the embryonic mouse telencephalon and found that apical contact is a characteristic feature of the cells that are part of a gap junction-coupled cluster. The throughput and versatility of the Autoinjector will render microinjection an accessible high-performance single-cell manipulation technique and will provide a powerful new platform for performing single-cell analyses in tissue for bioengineering and biophysics applications.
Bibliographical noteFunding Information:
We are grateful to the Services and Facilities of the Max Planck Institute of Molecular Cell Biology and Genetics for the outstanding support provided, notably J. Helppi and his team of the Animal Facility, J. Peychl and his team of the Light Microscopy Facility, and Jan Wagner and the MPI-CBG mechanical workshop. We would like to thank Prof. Dr. med. Pauline?Wimberger and Dr.?med. Nannette Gr?bling from the Klinik und Poliklinik f?r Frauenheilkunde und Geburtshilfe, Universit?tsklinikum Carl Gustav Carus of the Technische Universit?t Dresden, for providing fetal human tissue, and Michael Heide and Katie Long for dissecting and preparing the human fetal brain tissue. We would like to thank the Dr. Svante P??bo's entire team for helpful discussions, and Mareike Albert, Katie Long, and Nicola Maghelli for critical reading of the article and for their helpful comments. SBK acknowledges funds from the Mechanical Engineering department, College of Science and Engineering, MnDRIVE RSAM initiative of the University of Minnesota, McGovern Institute Neurotechnology (MINT) fund, National Institutes of Health (NIH) 1R21NS103098-01. GS was supported by NSF IGERT training fellowship, and the NSF IGERT travel award. ET was supported by NOMIS Distinguished Scientist Award for Dr. Svante P??bo. WBH was supported by grants from the DFG (SFB 655, A2), the ERC (250197), and ERA-NET NEURON (MicroKin).
- brain development
- computer vision
- neural stem cells
- single cell manipulation
PubMed: MeSH publication types
- Journal Article
- Research Support, N.I.H., Extramural
- Research Support, Non-U.S. Gov't