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EMBO Rep. 2019 Oct 4;20(10):e47880. doi: 10.15252/embr.201947880. Epub 2019 Aug 30.

Robotic platform for microinjection into single cells in brain tissue.

Author information

1
Department of Biomedical Engineering, University of Minnesota, Twin Cities, MN, USA.
2
Department of Biomedical Engineering, Duke University, Durham, NC, USA.
3
Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany.
4
Department of Mechanical Engineering, University of Minnesota, Twin Cities, MN, USA.
5
Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany.

Abstract

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.

KEYWORDS:

brain development; computer vision; neural stem cells; robotics; single cell manipulation

PMID:
31469223
DOI:
10.15252/embr.201947880
Free PMC Article

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