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An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry
JoVE 杂志
生物工程
This content is Free Access.
JoVE 杂志 生物工程
An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry

An Automated Method to Perform The In Vitro Micronucleus Assay using Multispectral Imaging Flow Cytometry

DOI:
10.3791/59324-v

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12:56 min

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May 13, 2019

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1Biology Department,Luminex Corporation

Chapters

  • 00:04Title
  • 00:33Clastogen and/or Aneugen Exposure
  • 01:49Sample Processing
  • 03:50Multispectral Imaging Flow Cytometry (MIFC)
  • 05:15Binucleated Cell (BNC) Identification
  • 06:48Micronucleus (MN) Identification
  • 07:53Polynucleated Population Identification
  • 08:41BNC and MN Mask Examination
  • 09:52Statistics Table Creation
  • 10:31Experimental File Batch Processing
  • 11:10Results: Representative Cell Scoring and Cytotoxicity Quantification
  • 12:23Conclusion

Summary

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The in vitro micronucleus assay is a well-established method for evaluating genotoxicity and cytotoxicity but scoring the assay using manual microscopy is laborious and suffers from subjectivity and inter-scorer variability. This paper describes the protocol developed to perform a fully automated version of the assay using multispectral imaging flow cytometry.

Tags

Keywords: In Vitro Micronucleus AssayMultispectral Imaging Flow CytometryAutomated MethodGenotoxicityCytotoxicityClastogensAneugensCell ExposureCytochalasin BPotassium ChlorideFormalinHoechst 33342High-throughputAutomated MicroscopyConventional Flow Cytometry

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