Lung Injury Evaluation via IHC and Histopathology

Paraffin sections (4 μm) of formalin fixed right upper lobe tissue were used for H&E staining, and immunohistochemistry with 1:250 mouse anti-human iNOS (BD Biosciences, USA) or no antibody (negative control) (22 (link)). Blinded slides for iNOS staining had 10 random regions/animal (40X on Zeiss Axioskop 40) manually counted and scored as 0 = no positive cells, 1 = occasional positive cells, 2 = large number of positive cells iNOS positive cells were primarily in airspaces. H&E stained slides of lungs were blinded and evaluated for airway epithelial injury, edema, hemorrhage, and inflammation (0–2 points each) (21 (link)).

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Hillman N.H., Abugisisa L., Royse E., Fee E., Kemp M.W., Kramer B.W., Schmidt A.F., Salomone F., Clarke M.W., Musk G.C, & Jobe A.H. (2020). Dose of budesonide with surfactant affects lung and systemic inflammation after normal and injurious ventilation in preterm lambs. Pediatric research, 88(5), 726-732.

Publication 2020

mouse anti-human iNOS Axioskop 40

Animal Antibody Cells Edema Formalin Hemorrhage Human Immunohistochemistry Inflammation Injury Inos Lungs Mouse Paraffin Tissue

Corresponding Organization : Saint Louis University

Other organizations : University of Western Australia, Maastricht University, University of Miami, Chiesi (Italy), Cincinnati Children's Hospital Medical Center, University of Cincinnati

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Variable analysis

independent variables

Presence or absence of primary antibody (mouse anti-human iNOS) during immunohistochemistry

dependent variables

Scoring of iNOS positive cells (0 = no positive cells, 1 = occasional positive cells, 2 = large number of positive cells)
Evaluation of airway epithelial injury, edema, hemorrhage, and inflammation (0–2 points each) based on H&E stained slides

control variables

Paraffin sections (4 μm) of formalin fixed right upper lobe tissue
Lung tissue from the same animal/specimen

positive control

No antibody (negative control) for immunohistochemistry

negative control

No antibody (negative control) for immunohistochemistry

Annotations

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