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In vivo Clonal Tracking of Hematopoietic Stem and Progenitor Cells Marked by Five Fluorescent Proteins using Confocal and Multiphoton Microscopy
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Journal JoVE Biologie
In vivo Clonal Tracking of Hematopoietic Stem and Progenitor Cells Marked by Five Fluorescent Proteins using Confocal and Multiphoton Microscopy

In vivo Clonal Tracking of Hematopoietic Stem and Progenitor Cells Marked by Five Fluorescent Proteins using Confocal and Multiphoton Microscopy

DOI:
10.3791/51669-v

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17:08 min

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August 06, 2014

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, ,
1Light Microscopy Core Facility,NHLBI/NIH, 2Hematology Branch,NHLBI/NIH

Chapitres

  • 00:05Titre
  • 02:28Mouse Cell Collection, Purification, and Transduction
  • 06:05Mouse Bone Marrow Transplantation
  • 07:45Setup for Confocal and Two-photon Microscopy
  • 10:12Imaging Sternum Bone Marrow Using Confocal and 2-photon Microscopy
  • 12:014D Time-lapse Imaging
  • 13:31Reconstruction and Analysis of 3D Volumes and 4D Time Sequences
  • 14:33Representative 3D Reconstructions of Sternal Bone Marrow and Other Organs
  • 16:17Conclusion

Summary

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Traduction automatique

Combinatorial 5 fluorescent proteins marking of hematopoietic stem and progenitor cells allows in vivo clonal tracking via confocal and two-photon microscopy, providing insights into bone marrow hematopoietic architecture during regeneration. This method allows non-invasive fate mapping of spectrally-coded HSPCs-derived cells in intact tissues for extensive periods of time following transplantation.

Tags

Keywords: Clonal TrackingHematopoietic Stem And Progenitor CellsFluorescent ProteinsConfocal MicroscopyMultiphoton MicroscopyLentiviral VectorsBone Marrow TransplantCell Fate MappingIn Vivo Hematopoiesis

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