Intravital laser confocal microscopy of pulmonary edema resulting from intestinal ischemia-reperfusion injury in the rat.
OBJECTIVE
To observe pulmonary edema resulting from intestinal ischemia-reperfusion injury. We used a newly developed laser confocal microscope to observe the subpleural capillary network and the superficial alveoli under intravital conditions, and created three-dimensional images of the pulmonary microcirculation to analyze the time course and spatial pattern of pulmonary exudative changes during intestinal ischemia-reperfusion injury in vivo.


METHODS
Prospective, randomized, unblinded study.


METHODS
Laboratory of a university hospital.


METHODS
Male Sprague-Dawley rats.


METHODS
The rats were injected intravenously with bovine serum albumin labeled with fluorescein isothiocyanate and subjected to 60 mins of intestinal ischemia, followed by 180 mins of reperfusion. During mechanical ventilation, the upper lobe of the right lung was examined in the intravital state using a high-speed confocal fluorescence microscope.


RESULTS
Interstitial edema and alveolar leakage were recognized as changes of interstitial fluorescence in the subpleural capillary network and as changes of alveolar fluorescence in the alveolar cross-sectional view. Although exudative changes in the interstitium and alveoli were observed during intestinal ischemia, there was a marked increase in both interstitial edema and alveolar leakage after intestinal reperfusion.


CONCLUSIONS
We observed pulmonary edema under intravital conditions and demonstrated the utility of a newly developed laser confocal microscope. This system not only enabled us to analyze the development of pulmonary edema three-dimensionally, but also allowed us to evaluate the pulmonary microcirculation.
Mitsuoka
H
H
Second Department of Surgery, Hamamatsu University School of Medicine, Japan.
Sakurai
T
T
Unno
N
N
Kaneko
H
H
Suzuki
S
S
Nakamura
S
S
Baba
S
S
Terakawa
S
S
D000704:Analysis of Variance; D000818:Animals; D007422:Intestines / Q000098:blood supply; D008297:Male; D008833:Microcirculation; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence; D009102:Multiple Organ Failure / Q000209:etiology / Q000503:physiopathology; D011446:Prospective Studies; D011650:Pulmonary Alveoli / Q000098:blood supply*; D011652:Pulmonary Circulation; D011654:Pulmonary Edema / Q000209:etiology* / Q000473:pathology* / Q000503:physiopathology; D011897:Random Allocation; D051381:Rats; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000150:complications* / Q000503:physiopathology; D012128:Respiratory Distress Syndrome / Q000473:pathology