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MitoMoby Mitochondrial Morphology Analysis Suite

Details

Project TitleMitoMoby Mitochondrial Morphology Analysis Suite
Track CodeP13108
Short Description

High Content Microscopic Assessment of Mitochondrial Morphology and Membrane Potential Using Morphological Binning

Abstract

MitoMoby is a high throughput and high content microscopic method for the analysis of mitochondrial morphology (MM) and membrane potential. MitoMoby delineates a procedure for MM analysis using the R platform for data analysis, annotation, and image registration and GE INCell Investigator (specifically, the GE INCell Developer Toolbox package) platform for image analysis.

Video Link: Quantitative Analysis of Mitochondrial Morphology

         

Seven measures of morphological analysis are reported in MitoMoby:  total mitochondrial object area (comprised of all phenotypes, representative of total mitochondrial mass), puncta area, rod area, networked area, large & round area, total mitochondrial object count, and the average fiber length (length along all branches) of networked mitochondrial objects.

     

Analysis of mitochondrial morphology and membrane potential under experimental conditions is achieved via the following steps:

  1. Image acquisition via wide-field fluorescence microscopy (using the GE INCell Analyzer 2000 Instrument)
  2. Image analysis (using GE INCell Developer Toolbox) including:
    1. Preprocessing  (2-D deconvolution and intensity normalization)
    2. Segmentation, (identification of mitochondrial objects);
    3. Classification into 4 morphological subtypes, (puncta, rods, networks, and large & round);
    4. Color definition using a machine learning-based classifier;
    5. Calculation of morphological measures such as area, count, and length;
    6. Quality control to eliminate small, dim objects
  3. Data annotation with experimental meta-data (treatment, concentration, plate, etc.) to facilitate unsupervised and supervised exclusion of low-quality cells and microscopic fields (out of focus, poorly stained) (using R and Excel);
  4. Data graphing and analysis (using R); and
  5. Statistical analysis (using R, Excel, and GraphPad).

    

Figure 1: Overview of morphological binning-based analysis of mitochondrial morphology.

    

Figure 2: Example of mitochondrial morphological assessment following treatment with t-butyl hydroperoxide (TBHP) at concentrations indicated. *p<0.05 v. vehicle

              

This method may be used to diagnose mitochondrial diseases involving dysregulation of morphological machinery or mitochondrial function. It can also be used as a research tool to investigate the regulation of mitochondrial dynamics. Most suitably, researchers can use this tool to assess pharmacological/toxicological evaluation of compound libraries or evaluate mitochondrial health in a large patient sample.

           

MitoMoby is presented in a ZIP file which contains the following files: R files, an XEAP file, an XAQP file, Microsoft Comma Separated Values files, a Microsoft Excel worksheet, a Text file, and a detailed instruction manual which delineates how to assemble and run the program.

          

Please note, FRD is only currently able to license to U.S. based entities.

    

Commercial Licenses are available by contacting FRD directly.  

        

For Commercial Licensing inquiries, please contact: 

       

Christine Dixon Thiesing

Licensing Officer

dixonthi@musc.edu

843.876.2260

 
TagsMUSC, MUSC Foundation for Research Development, Medical University of South Carolina, mitochondrial morphology, mitochondrial membrane potential, image analysis, cell analysis, cell imaging, InCell, morphological binning
 
Posted DateJan 6, 2015 12:19 PM

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Files

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Offerings

Name Price
Non-Commercial License MitoMoby Mitochondrial Morphology Analysis Suite 600.00 USD Buy
Commercial License MitoMoby Mitochondrial Morphology Analysis Suite 600.00 USD Buy