After After
Before Before

10164 - H&E, Centronuclear Myopathy

DEVELOPED FOR MEDICAL COLLEGE OF WISCONSIN

Centronuclear myopathy is a disease group characterized by the location of nuclei in the center of skeletal muscle fibers instead of the normal peripheral location. Symptoms of the disease include muscle weakness and diminished muscle tone.

This APP analyzes H&E stained skeletal muscle tissue to detect muscle fibers and nuclei located centrally within the fibers. The APP calculates the proportion of fibers with centronuclear myocytes.

KEYWORDS
H&E, Centronuclear Myopathy, Skeletal Muscle, Image Analysis, Digital Pathology

METHODS
The APP consists of two protocols. The first protocol excludes non-myocytes (e.g. connective tissue) from the subsequent analysis. The second protocol detects and segments myocytes and nuclei. The myocytes are identified based on the H&E color band in VIS. A myocyte candidate is defined as an actual myocyte if it passes certain morphological measures based on circularity and area. Nuclei candidates are defined as actual centrally located nuclei if they are fully surrounded by detected myocytes.

QUANTITATIVE OUTPUT VARIABLES
The output variables obtained from this protocol are:

  • Total number of fibers

  • Total number of internal nuclei

  • Internal nuclei proportion

  • Area of fibers (µm²)

  • Minor axis length (µm²)

AUXILIARY APPs
An auxiliary APP “01 Tissue Detection” is included for automatic detection of the tissue in the slide.

WORKFLOW
Step 1: Load and run the APP "01 Tissue Detection" for detection of the tissue in the slide
Step 2: Load and run the APP "02 Bundle Detection" for detection of bundles in the tissue
Step 3: Load and run the APP “03 Central Nuclei Fibers” for detection of muscle fibers and central nuclei

STAINING PROTOCOL
There is no staining protocol available.

ADDITIONAL INFORMATION 
The APP utilizes the Visiopharm EngineTM and Viewer software modules, where EngineTM offers an execution platform to expand processing capability and speed of image analysis. The Viewer allows a fast review together with several types of image adjustment properties e.g. outlining of regions, annotations and direct measures of distance, curve length, radius, etc. 

By adding the AuthorTM module the APP can be customized to fit other purposes. AuthorTM offers a comprehensive and dedicated set of tools for creating new fit-for-purpose analysis APPs, and no programming experience is required.

 

RUO
FIGURE 1
FIGURE 1
The relevant tissue of the whole slide image is automatically detected using “01 Tissue Detection”.
FIGURE 2
FIGURE 2
The muscle fibers are arranged in bundles separated by connective tissue. The protocol “02 Bundle Detection” automatically exclude the connective tissue from further analysis.
FIGURE 3
FIGURE 3
H&E stained skeletal muscle fibers. Some of the fibers have peripheral nuclei and some have central nuclei.
FIGURE 4
FIGURE 4
Detection of muscle fibers (yellow) and central nuclei (blue). Peripheral nuclei are included in the muscle fibers (yellow).