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Related Concept Videos

Mass Spectrometry: Complex Analysis01:21

Mass Spectrometry: Complex Analysis

Mass spectrometry is an important technique for the identification of pure compounds. However, it has some limitations for the analysis of complex mixtures, often due to excessive fragmentation making the spectrum too complicated to decipher. Mass spectrometry can be combined with suitable separation methods in sequence, forming hyphenated methods, which are useful in the analysis of complex mixtures.
GC–MS is a powerful hyphenated method commonly used in forensics and environmental...
Tandem Mass Spectrometry01:21

Tandem Mass Spectrometry

Tandem mass spectrometry is a technique that uses multiple mass analyzers in series to obtain a higher selectivity and reduce chemical noise during analyte detection. Instruments with multiple analyzers separated by an interaction cell enable secondary fragmentation and selected study of the fragment ions.Secondary fragmentations occur in the interaction cell and can be induced by various factors. Fragmentation induced by collision with inert gases, such as N2, Ar, He, etc., is called...

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Quantifying Synapses: an Immunocytochemistry-based Assay to Quantify Synapse Number
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Mass Synaptometry: Applying Mass Cytometry to Single Synapse Analysis.

Chandresh R Gajera1, Rosemary Fernandez2, Nadia Postupna3

  • 1Department of Pathology, Stanford University Medical Center, Stanford, CA, USA. gajera@stanford.edu.

Methods in Molecular Biology (Clifton, N.J.)
|January 31, 2022
PubMed
Summary
This summary is machine-generated.

Mass Synaptometry, a novel CyTOF approach, enables high-dimensional analysis of individual human synaptosomes. This method overcomes limitations of conventional flow cytometry for studying neurodegenerative and neurocognitive disorders.

Keywords:
CyTOF (Cytometry by Time of Flight)HumanMass SynaptometryMass cytometrySynTOF (Synapse by Time of Flight)SynapseSynaptosome

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Area of Science:

  • Neuroscience
  • Biochemistry
  • Mass Spectrometry

Background:

  • Synaptic degeneration is a key feature in neurodegenerative diseases like Alzheimer's and Parkinson's, and neurocognitive disorders such as schizophrenia and autism.
  • Conventional flow cytometry using fluorescence probes is limited in its ability to comprehensively analyze synaptosomes due to multiplexing constraints.
  • A broad range of probes is needed to assess synapse type, pathological proteins, and stress markers, exceeding current capabilities.

Purpose of the Study:

  • To introduce and describe Mass Synaptometry, a new method utilizing Cytometry by Time-Of-Flight mass spectrometry (CyTOF) for high-dimensional analysis of individual human synaptosomes.
  • To overcome the multiplexing limitations inherent in conventional fluorescence-based flow cytometry for synaptosome analysis.
  • To detail the methodology for preparing synaptosomes, generating and quality-controlling the SynTOF antibody panel, and optimizing CyTOF acquisition and analysis.

Main Methods:

  • Preparation of synaptosomes from human and mouse brain tissue.
  • Development and quality control of a specialized antibody panel (SynTOF) for mass spectrometry-based analysis.
  • Application of Cytometry by Time-Of-Flight mass spectrometry (CyTOF) for high-dimensional single-synaptosome analysis, including staining protocols and data acquisition/processing.

Main Results:

  • Successful application of CyTOF to high-dimensional analysis of individual human synaptosomes, termed Mass Synaptometry.
  • Overcoming multiplexing limitations of conventional flow cytometry for comprehensive synaptosome profiling.
  • Establishment of a robust protocol for synaptosome preparation, antibody panel generation, and CyTOF data analysis.

Conclusions:

  • Mass Synaptometry offers a powerful, high-dimensional approach for detailed analysis of individual synaptosomes.
  • This method significantly expands the capacity for studying synaptic pathology in neurodegenerative and neurocognitive disorders.
  • The described protocol provides a foundation for advancing research into the molecular mechanisms underlying brain disorders.