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Digestion of Whole Mouse Eyes for Multi-Parameter Flow Cytometric Analysis of Mononuclear Phagocytes
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JoVE 杂志 免疫与感染
Digestion of Whole Mouse Eyes for Multi-Parameter Flow Cytometric Analysis of Mononuclear Phagocytes

Digestion of Whole Mouse Eyes for Multi-Parameter Flow Cytometric Analysis of Mononuclear Phagocytes

DOI:
10.3791/61348-v

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09:58 min

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June 17, 2020

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, ,
1Department of Ophthalmology, Feinberg School of Medicine,Northwestern University, 2Department of Medicine, Division of Rheumatology, Feinberg School of Medicine,Northwestern University

Chapters

  • 00:05Introduction
  • 01:13Digestion of Ocular Tissue
  • 03:34Preparation of Cell Suspension
  • 05:19Staining of Cell Suspension for Mononuclear Phagocytes
  • 06:49Results: Mononuclear Phagocyte Populations in the Mouse Eye
  • 09:26Conclusion

Summary

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This protocol provides a method to digest whole eyes into a single cell suspension for the purpose of multi-parameter flow cytometric analysis in order to identify specific ocular mononuclear phagocytic populations, including monocytes, microglia, macrophages, and dendritic cells.

Tags

DigestionMouse EyesMononuclear PhagocytesFlow CytometryMacrophagesMonocytesMicrogliaDendritic CellsCell Surface MarkersChoroidal NeovascularizationDiabetic RetinopathyExperimental Autoimmune UveitisLupusCytometer ConfigurationDissectionMechanical Disruption

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