measuring fat and vacuolar space with digital pathology frank voelker, dvm, dacvp principle...
TRANSCRIPT
Measuring Fat and Vacuolar Space with Digital Pathology
Frank Voelker, DVM, DACVPPrinciple Pathologist
Trevor Johnson, MS Director, Image Analysis
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Originally An Algorithm for Measuring Fat
White Fat
Brown Fat
Lipid vacuoles in a variety of tissues (eg. Fatty liver or adrenal cortex)
Tissues composed of fibers or tubules (eg. skeletal muscle)
Tissues with hollow spaces or glandular lumena (eg. lung or thyroid gland)
Cerebral edema, demyelinization
Vacuoles in various cell types (eg. Lymphocytes)
What was actually needed: An algorithm for measuring circumscribed clear spaces in tissue
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Algorithm Requirements
Percent area of vacuoles or circumscribed spaces.
Total number of spaces
Mean area/diameter/perimeter of spaces
Histograms of areas, diameters or perimeters
Data Output:
Shape and Area Threshold ControlsHue Sensitivity AdjustmentMembrane Estimation or Completion
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Vacuole Algorithm Settings
Input and Output Windows
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How the Image is Analyzed
Define the tissue intensity
Determine the maximum and minimum vacuole areas
Determine how close each vacuole is allowed to resemble a circle and how tightly they are packed
Determine if Vacuole Perimeter Estimation (VPM) is needed
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Measuring White Fat Cells
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Measuring White Fat
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Measuring White Fat – Removing Small and Elongated Vacuoles
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Measuring White Fat – Removing Small Vacuoles
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Morphometric Considerations
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Remove Tissue (Just View Vacuole Spaces in Green)
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Remove Tissue (Parameter Too Low Including Tissue)
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Include Small Vacuoles
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Exclude Large Vacuoles
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Vacuole Analysis Fatty Liver
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Fatty Liver
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Measuring Brown Fat Vacuoles
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Measuring Brown Fat Vacuoles
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Measuring Brown Fat with Nuclear Counting
173 Nuclei
Area/No. Nuclei = Cell Size
No. Nuclei/No. Fat Droplets = No. Droplets/Cell
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Measuring Pulmonary Alveoli
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Measuring Pulmonary Alveoli
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Adrenal Gland
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Vacuole Analysis Adrenal
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Measuring Adrenal Vacuoles with Nuclear Counts
187 Nuclei
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Measuring Adrenal Vacuoles
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Measuring Bone Marrow Vacuoles
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Measuring Bone Marrow Vacuoles
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Measuring Skeletal Muscle Fibers
Type 1- Red Muscle. Slow twitch postural type muscle fiber. Aerobic oxidation.
Type 2- White Muscle Fast twitch fibers. ATPase +. Anaerobic oxidation
DenervationStatin-induced rhabdomyolysisNoninflammatory myopathiesCongential myopathiesSports physiology
Muscle fiber alterations are important in:
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Measuring Skeletal Muscle Fibers
In H&E sections, skeletal muscle fibers are poorly demarcated
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Measuring Skeletal Muscle Fibers
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Muscle Fiber Histogram
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Summary
An algorithm designed to measure vacuoles or circumscribed clear spaces in tissue was described in this presentation. The algorithm has proved useful in measuring clear spaces in a variety of tissues including white and brown fat, skeletal muscle fibers, pulmonary alveoli, fatty liver and adrenal cortex. Data outputs included percent as well as histograms and mean areas, diameters or perimeters of vacuoles or clear areas within tissue.