Protocol for the application of single-cell damage in murine intestinal organoid models

Research output: Contribution to journal › Article › Research › peer review

Authors

  • Anna Elisabeth Seidler
  • Sören Donath
  • Lara Gentemann
  • Manuela Buettner
  • Alexander Heisterkamp
  • Stefan Kalies

Research Organisations

External Research Organisations

  • NIFE - Lower Saxony Centre for Biomedical Engineering, Implant Research and Development
  • REBIRTH Research Center for Translational Regenerative Medicine
  • Hannover Medical School (MHH)
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Details

Original languageEnglish
Article number103153
Number of pages31
JournalSTAR Protocols
Volume5
Issue number3
Early online date30 Jul 2024
Publication statusPublished - 20 Sept 2024

Abstract

Spatially defined organoid damage enables the study of cellular repair processes. However, capturing dynamic events in living tissues is technically challenging. Here, we present a protocol for the application of single-cell damage in intestinal organoid models. We describe steps for isolating and cultivating murine colon organoids, lentivirus generation and transduction of organoids, single-cell ablation by a femtosecond laser, and follow-up imaging analysis. We provide examples for the image acquisition pipeline of dynamic processes in organoids using a confocal microscope. For complete details on the use and execution of this protocol, please refer to Donath et al.1,2

Keywords

    Biophysics, Microscopy, Molecular Biology, Organoids, Single Cell

ASJC Scopus subject areas

Cite this

Protocol for the application of single-cell damage in murine intestinal organoid models. / Seidler, Anna Elisabeth; Donath, Sören; Gentemann, Lara et al.
In: STAR Protocols, Vol. 5, No. 3, 103153, 20.09.2024.

Research output: Contribution to journal › Article › Research › peer review

Seidler, AE, Donath, S, Gentemann, L, Buettner, M, Heisterkamp, A & Kalies, S 2024, 'Protocol for the application of single-cell damage in murine intestinal organoid models', STAR Protocols, vol. 5, no. 3, 103153. https://doi.org/10.1016/j.xpro.2024.103153
Seidler, A. E., Donath, S., Gentemann, L., Buettner, M., Heisterkamp, A., & Kalies, S. (2024). Protocol for the application of single-cell damage in murine intestinal organoid models. STAR Protocols, 5(3), Article 103153. https://doi.org/10.1016/j.xpro.2024.103153
Seidler AE, Donath S, Gentemann L, Buettner M, Heisterkamp A, Kalies S. Protocol for the application of single-cell damage in murine intestinal organoid models. STAR Protocols. 2024 Sept 20;5(3):103153. Epub 2024 Jul 30. doi: 10.1016/j.xpro.2024.103153
Seidler, Anna Elisabeth ; Donath, Sören ; Gentemann, Lara et al. / Protocol for the application of single-cell damage in murine intestinal organoid models. In: STAR Protocols. 2024 ; Vol. 5, No. 3.
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