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Surgical model for functional intravital microscopy of the lung using an acute window
Author(s) -
Hanna Gabi,
Palmer Gregory,
Shan Siqing,
Radiloff Daniel,
Dewhirst Mark,
McMahon Timothy,
Irwin David,
Schroeder Thies
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.lb640
Subject(s) - intravital microscopy , lung , medicine , blood flow , biomedical engineering , magnification , functional imaging , pathology , radiology , microcirculation , computer science , computer vision
Introduction Numerous pathologies of the lung could benefit from a model of functional microscopic imaging of the lung in laboratory animals. Recent developments in window chamber microscopy enable us to measure key parameters of the lung including capillary blood flow and hemoglobin saturation. Functional intravital microscopy at high magnification, however, requires a still image for extended exposure times which is very difficult to obtain in the lung due to respiratory and cardiac movement. Methods Female Sprague Dawley rats were anesthetized with ketamine/xylazine, tracheotomized, connected to a ventilator and subjected to open‐chest surgery. A customized window was protruded into the surgical opening and sutured to the outside of the wound sealing the chest cavity resulting in direct contact to the lung. Vital signs were assessed using MouseOx. Imaging was done using a fluorescence Zeiss microscope at 100 x magnification. Image analysis was done using Matlab and Image J. Results We were able to eliminate lateral movement of the lung due to respiration and heart beat, and observe blood flow in vivo over a period of >1 min. We were able to measure hemoglobin saturation in pulmonary capillaries Conclusions Our surgical model enables intravital microscopy of the lung over extended imaging times. This model has clear potential to serve as a standard platform to study pathophysiologies of the lung.