Skeletal Muscle Fiber Analysis (Reticulin Stain) (Basic)

Skeletal Muscle Fiber Analysis (Reticulin Stain) (Advanced)

IQ-BOT-0011

Details

Name
Skeletal Muscle Fiber Analysis (Reticulin Stain)
IQbot ID
IQBOT-0011
Version
1.0.0
Research Area(s)
Arthritis & Osteoarthritis, Drug Toxicity & Metabolism, Orthopedics, & Common Tools
Processing Time Expectation
Less than 2 Minutes per Image
Acquisition Hardware
Wide-Field, Optical Microscope in Bright-Field (Transmission) Mode Using a Color Mosaic CCD or Monochrome CCD with RGB Filter
Techniques, Dyes, Stains, & Proteins
Reticulin Stain
Sample Context
In Tissue
Name
Skeletal Muscle Fiber Analysis (Reticulin Stain)
IQbot ID
IQBOT-0011
Version
1.0.0
Research Area(s)
Arthritis & Osteoarthritis, Drug Toxicity & Metabolism, Orthopedics, & Common Tools
Processing Time Expectation
Less than 2 Minutes per Image
Acquisition Hardware
Wide-Field, Optical Microscope in Bright-Field (Transmission) Mode Using a Color Mosaic CCD or Monochrome CCD with RGB Filter
Techniques, Dyes, Stains, & Proteins
Reticulin Stain
Sample Context
In Tissue

Output Parameters

Total Fibrils
The total number of segmented fibrils in the image.
Mean Area (pixels)
The mean area of segmented fibrils.
Mean Aspect Ratio
The mean aspect ratio (ratio between the major and minor axes of the best fit ellipse) of segmented fibrils.
Mean Roundness
The mean roundness (perimeter2/(4π*area)) of segmented fibrils (a measure of fibril border roughness or tortuosity).
Mean Diameter (pixels)
The mean diameter (the average length of diameters passing through the fibril's centroid measured at 2° intervals) of segmented fibrils.
Mean Perimeter (pixels)
The mean perimeter (the length of fibril border) of segmented fibrils.

 

This section will identify any output parameters that are only output by the IQbot in its advanced configuration.

 

Total Fibrils
The total number of segmented fibrils in the image.
Mean Area (pixels)
The mean area of segmented fibrils.
Mean Aspect Ratio
The mean aspect ratio (ratio between the major and minor axes of the best fit ellipse) of segmented fibrils.
Mean Roundness
The mean roundness (perimeter2/(4π*area)) of segmented fibrils (a measure of fibril border roughness or tortuosity).
Mean Diameter (pixels)
The mean diameter (the average length of diameters passing through the fibril's centroid measured at 2° intervals) of segmented fibrils.
Mean Perimeter (pixels)
The mean perimeter (the length of fibril border) of segmented fibrils.
Area (pixels)
The area of each fibril segmented.
Aspect Ratio
The aspect ratio (ratio between the major and minor axes of the best fit ellipse) of each fibril segmented.
Roundness
The roundness (perimeter2/(4π*area)) of each fiber segmented (a measure of fibril border roughness or tortuosity).
Diameter (pixels)
The diameter (the average length of diameters passing through the fibril's centroid measured at 2° intervals) of each fibril segmented.
Perimeter (pixels)
The perimeter (the length of fibril border) of each fibril segmented

Image Acquisition


This IQbot was developed to analyze images acquired using a wide-field, optical microscope in bright-field (transmission) mode using a color mosaic CCD or monochrome CCD with RGB filter. Sections imaged and subsequently submitted for analysis should be treated with a reticulin stain and depict skeletal muscle fiber networks.

 
 

The Skeletal Muscle Fiber Analysis (Reticulin Stain) IQbot has been built to accommodate the following imaging types only.

 24-bit RGB
 .tif
  .tiff
  .bmp
 .jpg
.jpeg
 
 

As we reference for IQbots in general here, the Skeletal Muscle Fiber Analysis (Reticulin Stain) is only ultimately effective if you upload high quality images for processing. Please ensure that any images you upload follow the below guidelines specific to this IQbot.

High-Resolution:  The quality of the output data you receive is largely dependent on the input image resolution. Please provide high-resolution images to ensure that the IQbot can most effectively differentiate between features.
In-Focus:  Please produce the most in-focus images possible for analysis. Images with high background contrast will result in more accurate and useful data and overlays.
Properly Saturated:  If your submitted images are significantly undersaturated or oversaturated, it is possible that the IQbot will be unable to detect structures, or may group structures together. Please ensure that the images you upload are saturated appropriately.
Clear of Artifacts:  Please take care to avoid creating bubbles, debris, or otherwise inducing artifacts into your image. Such additional features may corrupt the resulting data.
Clear of Accessary Information:  Please do not submit images retaining any annotations, legends, timestamps, or other markings. Every part of the images that you upload will be analyzed and included in the output data.
 
 
 

If you are uncertain whether or not your images meet the quality standards for which this IQbot was validated, or have additional questions relating to image quality, please view our FAQs or contact the ImageQuantify.com support team.

 

     (216) 678-9258           help@imagequantify.com

 

Validation Information


This IQbot has been successfully validated for use, and a copy of its validation certificate accompanies the results of each order it analyzes. In accordance with general ImageQuantify.com validation procedures, this IQbot has been found to provide quantitative information equivalent to that of a manual observer, but with no variability and increased breadth of information.

 
 

The first step of the validation process was to establish the ground truth against which the IQbot’s efficacy would be compared. For the purpose of the Skeletal Muscle Fiber Analysis (Reticulin Stain) IQbot, ground truth was defined on a manual observer basis. Trained observers were guided through a standardized, semi-automated process to produce output data from a specially selected validation image set. This image set represented the entire range of expected output values (e.g., with high or low skeletal muscle fiber density). Images were presented for analysis in a randomized order and deidentified to protect against observer biases. The observer mean for each parameter measured was accepted as ground truth.

By analyzing the inter- and intraobserver variability present in the manual observation process, acceptable criteria were established so as to compare the IQbot’s output to that of a trained replacement observer. Ultimately, the IQbot’s performance proved to offer significant upgrades over manual or semi-automated analysis techniques, and validation was considered successful. Further considerations ensured that the IQbot rejected and flagged concerning image-specific outputs and performed within operation time constraints.

 

The Skeletal Muscle Fiber Analysis IQbot analyzes muscle fiber content and morphology in histological cross-sections of skeletal muscle stained for reticulin, a connective tissue fiber composed of type III collagen. A number of metabolic and connective tissue disorders can affect muscle fiber density, size, and shape. Unfortunately, such phenotypic effects can be subtle, necessitating the quantification of morphological parameters for as many muscle fibers as possible within a given image or cross-section for a large cohort of samples — an exhaustive process if performed manually. Fortunately, the Skeletal Muscle Fiber Analysis IQbot was specifically designed to rapidly and comprehensively segment muscle fibers (in cross-section) within an image, extracting fibril number, area, aspect-ratio or elongation, roundness or roughness, perimeter, and diameter. These can be presented as averages or, using an advanced version of the IQbot, can be reported for each and every fibril segmented. Additionally, segmented fibrils are pseudo-colored red and superimposed upon each image analyzed to enable verification of the results and segmentation algorithm.

DEMO IMAGE
IQBOT APPLIED IMAGE

 

SAMPLE DATA
Image Total Fibrils Mean Area
(pixels)
Mean Aspect Ratio Mean Roundness Mean Diameter
(pixels)
Mean Perimeter
(pixels)
Area
(pixels)
Aspect Ratio Roundness Diameter
(pixels)
Perimeter
(pixels)
Sample 1401 573.69 1.581 1.221 25.68 91.32 251 1.408 1.058 17.49 57.78
1400 Additional Rows 1400 Additional Rows 1400 Additional Rows 1400 Additional Rows 1400 Additional Rows