Fully Automated Microinjection System for Xenopus laevis Oocytes with Integrated Sorting and Collection

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 au...

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Bibliographic Details
Published inJournal of Laboratory Automation Vol. 16; no. 3; pp. 186 - 196
Main Authors Graf, Siegfried F., Madigou, Thierry, Li, Ruoya, Chesné, Christophe, Stemmer, Andreas, Knapp, Helmut F.
Format Journal Article
LanguageEnglish
Published Los Angeles, CA SAGE Publications 01.06.2011
Subjects
Animals
Automation, Laboratory - methods
Microinjections - instrumentation
Microinjections - methods
Oocytes - growth & development
Oocytes - metabolism
Protein Transport
Transfection - methods
Xenopus laevis
oocyte
oocyte self-orientation
Xenopus laevis
cell sorting
automated microinjection
vision based
automated collection
unlabeled sorting
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Summary: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 4 h for the same amount and delivers injected oocytes of reproducible quality and without the fatigue symptoms experienced during the manual approach.
ISSN:2211-0682
1535-5535
1540-2452
2211-0690
DOI:10.1016/j.jala.2011.03.006