Fully Automated Microinjection System for Xenopus laevis Oocytes with Integrated Sorting and Collection
| Autor: | Siegfried Graf, Christophe Chesné, Ruoya Li, Andreas Stemmer, Thierry Madigou, Helmut Knapp |
|---|---|
| Přispěvatelé: | Interactions cellulaires et moléculaires (ICM), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Centre National de la Recherche Scientifique (CNRS), Université de Rennes (UR)-Centre National de la Recherche Scientifique (CNRS) |
| Rok vydání: | 2011 |
| Předmět: |
MESH: Protein Transport
Microinjections Xenopus MESH: Automation Laboratory 02 engineering and technology MESH: Microinjections Transfection Bioinformatics MESH: Oocytes Xenopus laevis 03 medical and health sciences MESH: Xenopus laevis Animals MESH: Animals Microinjection 030304 developmental biology Automation Laboratory 0303 health sciences biology Vision based MESH: Transfection Sorting Cell sorting 021001 nanoscience & nanotechnology biology.organism_classification Computer Science Applications Cell biology Protein Transport Medical Laboratory Technology Fully automated Oocytes 0210 nano-technology [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology |
| Zdroj: | Journal of Laboratory Automation Journal of Laboratory Automation, 2011, 16 (3), pp.186-96. ⟨10.1016/j.jala.2011.03.006⟩ |
| ISSN: | 1535-5535 |
| DOI: | 10.1016/j.jala.2011.03.006 |
| Popis: | International audience; Microinjection is the most flexible transfection method in terms of choice of reagents to inject into cells. But this method lacks the high throughput to compete with less flexible methods like chemical- or viral-based approaches. Various approaches have been pursued to increase the throughput by automating the microinjection process. However, these approaches focused solely on the microinjection itself and disregarded the tasks before and after the injection, which also belong to the critical time path of the whole process, that is, sorting out viable cells from a cell suspension, placing the cell for injection, and collecting the cell after the injection. In the approach with our XenoFactor, we demonstrate a system capable of running the whole process automatically. By optimizing the XenoFactor for Xenopus laevis oocytes, we could demonstrate the successful automated injection. Starting from a suspension with a mixture of defolliculated oocytes at different stages and quality levels, the manual approach requires 1 day in total for the preparation of 400 microinjected oocytes. The XenoFactor takes only 4h for the same amount and delivers injected oocytes of reproducible quality and without the fatigue symptoms experienced during the manual approach. |
| Databáze: | OpenAIRE |
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