Measuring fat and vacuolar space with digital pathology
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Measuring Fat and Vacuolar Space with Digital Pathology. Frank Voelker, DVM, DACVP Principle Pathologist. Trevor Johnson, MS Director, Image Analysis. Originally An Algorithm for Measuring Fat. What was actually needed: An algorithm for measuring circumscribed clear spaces in tissue.

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Measuring Fat and Vacuolar Space with Digital Pathology

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Measuring fat and vacuolar space with digital pathology

Measuring Fat and Vacuolar Space with Digital Pathology

Frank Voelker, DVM, DACVP

Principle Pathologist

Trevor Johnson, MS

Director, Image Analysis


Originally an algorithm for measuring fat

Originally An Algorithm for Measuring Fat

What was actually needed: An algorithm for measuring circumscribed clear spaces in tissue

  • 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)


Algorithm requirements

Algorithm Requirements

  • Data Output:

  • Percent area of vacuoles or circumscribed spaces.

  • Total number of spaces

  • Mean area/diameter/perimeter of spaces

  • Histograms of areas, diameters or perimeters

  • Shape and Area Threshold Controls

  • Hue Sensitivity Adjustment

  • Membrane Estimation or Completion


Vacuole algorithm settings

Vacuole Algorithm Settings

Input and Output Windows


How the image is analyzed

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


Measuring white fat cells

Measuring White Fat Cells


Measuring white fat

Measuring White Fat


Measuring white fat removing small and elongated vacuoles

Measuring White Fat – Removing Small and Elongated Vacuoles


Measuring white fat removing small vacuoles

Measuring White Fat – Removing Small Vacuoles


Morphometric considerations

Morphometric Considerations


Remove tissue just view vacuole spaces in green

Remove Tissue (Just View Vacuole Spaces in Green)


Remove tissue parameter too low including tissue

Remove Tissue (Parameter Too Low Including Tissue)


Include small vacuoles

Include Small Vacuoles


Exclude large vacuoles

Exclude Large Vacuoles


Vacuole analysis fatty liver

Vacuole Analysis Fatty Liver


Fatty liver

Fatty Liver


Measuring brown fat vacuoles

Measuring Brown Fat Vacuoles


Measuring brown fat vacuoles1

Measuring Brown Fat Vacuoles


Measuring brown fat with nuclear counting

Measuring Brown Fat with Nuclear Counting

173 Nuclei

Area/No. Nuclei = Cell Size

No. Nuclei/No. Fat Droplets = No. Droplets/Cell


Measuring pulmonary alveoli

Measuring Pulmonary Alveoli


Measuring pulmonary alveoli1

Measuring Pulmonary Alveoli


Adrenal gland

Adrenal Gland


Vacuole analysis adrenal

Vacuole Analysis Adrenal


Measuring adrenal vacuoles with nuclear counts

Measuring Adrenal Vacuoles with Nuclear Counts

187 Nuclei


Measuring adrenal vacuoles

Measuring Adrenal Vacuoles


Measuring bone marrow vacuoles

Measuring Bone Marrow Vacuoles


Measuring bone marrow vacuoles1

Measuring Bone Marrow Vacuoles


Measuring skeletal muscle fibers

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

Muscle fiber alterations are important in:

Denervation

Statin-induced rhabdomyolysis

Noninflammatory myopathies

Congential myopathies

Sports physiology


Measuring skeletal muscle fibers1

Measuring Skeletal Muscle Fibers

In H&E sections, skeletal muscle fibers are poorly demarcated


Measuring skeletal muscle fibers2

Measuring Skeletal Muscle Fibers


Muscle fiber histogram

Muscle Fiber Histogram


Summary

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.


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