Lung Fixation and Histological Analysis

Following induction of isoflurane anesthesia, a midline abdominal incision was made, and the abdominal aorta was transected to exsanguinate and euthanize the animal. A midline incision was continued through the thoracic cavity and neck to allow injection of 5 ml of normal saline into the right ventricle of the heart to flush the pulmonary vasculature of residual blood. A luer lock–hubbed 20-gauge × ½-inch cannula was inserted into the trachea and secured in place with a suture. The lungs were fixed by insufflation with 10% neutral buffered formalin at 20 cm H₂O for 24 hours at room temperature. The lung lobes were removed and embedded in paraffin, and 5-μm tissue slices were prepared and stained with hematoxylin and eosin by standard histopathology techniques (22 (link)). Histology images were acquired with an Aperio ScanScope CS slide scanner (Leica Biosystems) using a 40× objective lens and subsequently analyzed using Aperio ImageScope software version 12.3 (Leica Biosystems).

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Li Y., Beitelshees M., Fang L., Hill A., Ahmadi M.K., Chen M., Davidson B.A., Knight P I.I.I., Smith RJ J.r., Andreadis S.T., Hakansson A.P., Jones C.H, & Pfeifer B.A. (2016). In situ pneumococcal vaccine production and delivery through a hybrid biological-biomaterial vector. Science Advances, 2(7), e1600264.

Publication 2016

Aperio ScanScope CS slide scanner Aperio ImageScope software version 12.3

Abdominal Abdominal aorta Anesthesia Animal Blood Cannula Embedded paraffin Eosin Flush Formalin Insufflation Isoflurane Lens Lung Neck Normal saline Right ventricle of heart Suture Thoracic cavity Tissue Trachea

Corresponding Organization : University at Buffalo, State University of New York

Other organizations : Center for Excellence in Education, Lund University

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Variable analysis

independent variables

Induction of isoflurane anesthesia
Transection of the abdominal aorta to exsanguinate and euthanize the animal
Injection of 5 ml of normal saline into the right ventricle of the heart to flush the pulmonary vasculature of residual blood
Insufflation of the lungs with 10% neutral buffered formalin at 20 cm H2O for 24 hours at room temperature

dependent variables

Histology of the lung tissue

control variables

Midline abdominal and thoracic incisions
Insertion of a luer lock–hubbed 20-gauge × ½-inch cannula into the trachea and securing it in place with a suture

positive controls

Not specified

negative controls

Not specified

Annotations

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