Webinar: Automation of the in vitro micronucleus assay using imaging flow cytometry

Webinar Title: Automation of the in vitro micronucleus assay using imaging flow cytometry

Date: Tuesday, October 24, 2017

Time: 10:00 AM CST

Duration: 60 minutes



 What Does it Cover?

The in vitro micronucleus assay is one of the most widely used tests to quantify genotoxicity and cytotoxicity, especially as a screening tool in the development of chemicals and pharmaceuticals. Micronuclei (MN) are formed from whole chromosomes or chromosome fragments that lag behind during the metaphase-anaphase transition and are excluded from the main nucleus following division. MN form into small, rounded bodies surrounded by their own nuclear envelope and represent chromosomal mutations that can be used as an endpoint in genotoxicity testing. Typically performed by manual microscopy, the MN assay is laborious and can be subject to scorer bias. To overcome this, automated microscopy and conventional flow cytometry methods have been developed. However, these methods suffer from several limitations such as the requirement to create high quality slides in the case of microscopy and the lack of visual confirmation of MN in the case of flow cytometry. The ImageStream®X (ISX) imaging flow cytometer has the potential to overcome these limitations as it combines the speed, statistical robustness and rare event capture capability of conventional flow cytometry with high resolution fluorescent imagery.

In this webinar, adaptation of the in vitro MN assay to an imaging flow cytometry-based method will be described. Using the ISX Mark II imaging flow cytometer, images of micronucleated mono- and binucleated cells as well as polynucleated cells can be captured at a high flow rate and automatically identified and scored in the Image Data Exploration and Analysis Software (IDEAS®) that accompanies the ISX. A data analysis template created specifically for this application allows for the determination of both genotoxicity and cytotoxicity following treatment with known clastogens and aneugens. This work is the first demonstration of fully automated method for performing the in vitro MN assay on an imaging flow cytometry platform.
 

 What Will You Learn?

  • Purpose of the in vitro MN assay and how/why it is performed
  • Limitations of currently available automated techniques (automated slide-scanning microscopy and conventional flow cytometry)
  • Sample preparation methods including treatment of TK6 cells with known clastogens/aneugens, sample processing protocols
  • Data collection on the ISX MKII
  • Description of the IDEAS® data analysis template to identify micronucleated mononucleated and binucleated cells as well as polynucleated cells
  • Determination of cytotoxicity and genotoxicity using statistical quantities extracted from the data analysis template

 Who Should Attend?

Anyone interested in learning more about the ImageStream imaging flow cytometer and its use in DNA damage assays.

 

Speaker Bio
Matthew A. Rodrigues, PhD
Research Scientist
Amnis-Merck KGaA, Darmstadt, Germany
Matthew Rodrigues is a Research Scientist with Amnis (part of MilliporeSigma). He earned his BSc in Physics from Laurentian University in Sudbury (Canada) in 2006, his MSc in Biomedical Physics from Ryerson University in Toronto (Canada) in 2008 and his PhD in Medical Physics from Carleton University in Ottawa (Canada) in 2015. His PhD research involved adapting the microscope-based micronucleus assay to an imaging flow cytometry technique for radiation biodosimetry.

Prior to joining Amnis, Matthew worked as Research Scientist in the Nuclear Emergency Preparedness and Response Division of Health Canada.