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Super-Resolution Imaging to Study Co-Localization of Proteins and Synaptic Markers in Primary Neurons
JoVE 杂志
神经科学
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JoVE 杂志 神经科学
Super-Resolution Imaging to Study Co-Localization of Proteins and Synaptic Markers in Primary Neurons

Super-Resolution Imaging to Study Co-Localization of Proteins and Synaptic Markers in Primary Neurons

DOI:
10.3791/61434-v

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14:02 min

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October 31, 2020

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, , , , , , , ,
1Department of Molecular Biochemistry and Pharmacology,Istituto di Ricerche Farmacologiche Mario Negri IRCCS, 2Department of Neuroscience,Istituto di Ricerche Farmacologiche Mario Negri IRCCS, 3Dulbecco Telethon Institute,Istituto di Ricerche Farmacologiche Mario Negri IRCCS

Chapters

  • 00:00Introduction
  • 01:04Primary Cultures
  • 01:48Immunofluorescence Staining
  • 03:32SIM Microscopy
  • 04:01Microscope Calibration
  • 07:11Acquisition
  • 09:24Post-production: Image Reconstruction
  • 11:24Colocalization with Profile Analysis
  • 12:05Quantification of Pearson’s and Mander’s
  • 12:27Results
  • 13:16Conclusions

Summary

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This protocol shows how to employ super-resolution microscopy to study protein co-localization in primary neuronal cultures.

Tags

Super-resolution ImagingCo-localizationPrimary NeuronsSynaptic MarkersStructured Illumination Microscopy (SIM)Neurodegenerative DisordersHippocampal NeuronsFixationImmunostainingMounting SolutionCoverglass

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