Novel applications of confocal microscopy in lung toxicology.
Antonini-J; Taylor-MD; Roberts-JR
FASEB J 2002 Mar 16(5):A738
Confocal microscopy (CM) allows optical sectioning of a sample without physical damage. We examined the deposition of inhaled asbestos and quantified lung injury after intratracheal amiodarone (AD) treatment with CM. Exposed lungs were: 1) airway fixed, stained with lucifer yellow (LY; 0.1 mg/ml), and embedded in plastic; or 2) inflated with ethidium homodimer (EH; 0.4 mM), cryopreserved, and sectioned. Images of asbestos deposited in human lung parenchyma were reconstructed in 3-D. Asbestos fiber orientation in situ was obtained while preserving anatomic relationships and fiber geometry. Chronic lung injury in rats was quantified by measuring the connective tissue area stained by LY as an indicator of alveolar wall thickening. AD induced a fibrotic response at 28 days posttreatment. For acute cytotoxicity, AD compromised the integrity of cell membranes as evidenced by a marked increase in the number of cells labeled with EH. Damage was more prevalent in the airways at 15 min, but by 3 days, the injury was localized to the parenchyma. CM can quantify toxic responses and identify locations of damage without physical disruption of tissue and may be useful in the analysis of human biopsy samples.
Microscopy; Toxicology; Asbestos-dust; Lung-disorders; Injuries; Asbestos-fibers; Alveolar-cells; Cytotoxicity; Cell-damage; Lung-tissue
National Institute for Occupational Safety and Health, 1095 Willowdale Road, M/S 2015, Morgantown, WV 26505
Abstract; Conference/Symposia Proceedings
Disease and Injury; Asthma and Chronic Obstructive Pulmonary Disease
The FASEB Journal, Experimental Biology 2002, New Orleans, Louisiana, April 20-24, 2002