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Workflow for High-content, Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data, Supported by Open Source Software
JoVE Journal
Biologia
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JoVE Journal Biologia
Workflow for High-content, Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data, Supported by Open Source Software

Workflow for High-content, Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data, Supported by Open Source Software

DOI:
10.3791/51882-v

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09:57 min

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December 16, 2014

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,
1Cancer Biology,UCL Cancer Institute

Capitoli

  • 00:05Titolo
  • 01:17Reverse Transfection siRNA Screening
  • 03:20Imaging and Image Segmentation
  • 05:21Data Extraction
  • 07:39Results: Representative Raw Individual Cell Data Processing
  • 09:27Conclusion

Summary

Traduzione automatica

Traduzione automatica

Presented is a flexible informatics workflow enabling multiplexed image-based analysis of fluorescently labeled cells. The workflow quantifies nuclear and cytoplasmic markers and computes marker translocation between these compartments. Procedures are provided for perturbation of cells using siRNA and reliable methodology for marker detection by indirect immunofluorescence in 96-well formats.

Tags

Keywords: Cell QuantificationFluorescent MarkersHigh-content MicroscopySingle-cell AnalysisCell HeterogeneityCell ProfilerOpen-source SoftwareSubcellular SegmentationSiRNA ScreenG1 Checkpoint RegulatorsCell PerturbationFluorescence-based Cellular Markers

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