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DetectSyn: A Rapid, Unbiased Fluorescent Method to Detect Changes in Synapse Density
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JoVE Journal Biochimica
DetectSyn: A Rapid, Unbiased Fluorescent Method to Detect Changes in Synapse Density

DetectSyn: A Rapid, Unbiased Fluorescent Method to Detect Changes in Synapse Density

DOI:
10.3791/63139-v

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09:10 min

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July 22, 2022

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, ,
1Wake Forest Translational Alcohol Research Center,Wake Forest School of Medicine, 2Physiology and Pharmacology,Wake Forest School of Medicine

Capitoli

  • 00:04Introduction
  • 00:34Primary Antibody Incubation
  • 01:29Sample Washes, Secondary Antibody Incubation, Ligation, Amplification, and Mounting
  • 03:49Digital Imaging with a Confocal Microscope
  • 05:49Image Analysis
  • 07:41Results: Increased Synapse Formation by Rapid Antidepressants in the Presence of GABAB Agonists
  • 08:42Conclusion

Summary

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DetectSyn is an unbiased, rapid fluorescent assay that measures changes in relative synapse (pre- and postsynaptic engagement) number across treatments or disease states. This technique utilizes a proximity ligation technique that can be used both in cultured neurons and fixed tissue.

Tags

Keywords: DetectSynFluorescent MethodSynapse DensityProximity Ligation AssayPrimary AntibodySecondary AntibodyLigationAmplificationMounting

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