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All morphometric studies were performed using a confocal microscope by two experienced technicians blinded to the treatment.
Serial sections (5 µm in thickness) were prepared, stained with Mayer's hematoxylin-eosin (H&E), and examined under microscope by two independent pathologists.
The slides were then read under a fluorescent microscope by two trained laboratory personnel.
The immunohistochemical staining was evaluated under a light microscope by two independent investigators.
AECIIs were examined under a transmission electron microscope by two blinded, independent observers.
The slides were examined under light microscope by two independent investigators with the experience of liver pathology.
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The intensity of staining and the percentage of stained cells were evaluated under a light microscope by three independent pathologists without knowledge of clinical data.
For cohort 1, the Ecd staining intensity was evaluated under a light microscope by six independent observers including three of our pathology collaborators, and expressed on a scale of 0 to 3: 0, no staining; 1, weak; 2, moderate; 3, strong.
To evaluate the IHC and CISH staining, random selections of slides were evaluated simultaneously on a multi-headed microscope by three pathologists and a consensus score was reported for each slide.
The HES-stained sections were reviewed under a microscope independently by two experienced pathologists (OAH, AMB) and classified according to histopathological type and grade according to the World Health Organization Classification of Tumours [ 23] and the Nottingham grading system [ 12, 33].
Specifically, from the toe we obtained four physes (two joints) and at least three sections per physis were analyzed by measuring the surface areas of 10 randomly chosen lacunae from each of three representative microscope views by two independent investigators (AFS and BP).
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