Published ahead of print on October 5, 2006, doi:10.1164/rccm.200605-684OC Am. J. Respir. Crit. Care Med., Volume 175, Number 1, January 2007, 22-31 A more recent version of this article appeared on January 1, 2007
Submitted on May 20, 2006 In Vivo Imaging of the Bronchial Wall Microstructure Using Fibered Confocal Fluorescence MicroscopyLuc Thiberville1*,1 Clinique Pneumologique, Rouen University Hospital, Rouen, France, 2 Pathology Department, Rouen University Hospital, Rouen, France, 3 Mauna Kea Technologies, Paris, France; INRIA Sophia Antipolis, Sophia Antipolis Cedex, France, 4 Mauna Kea Technologies, Paris, France, 5 CNRS UMR 7033, Universite Pierre et Marie Curie, Paris, France * To whom correspondence should be addressed. E-mail: luc.thiberville{at}univ-rouen.fr.
Rationale : Fibered confocal fluorescence microscopy (FCFM) is a new technique that produces microscopic imaging of a living tissue, through a 1mm fiberoptic probe that can be introduced into the working channel of the bronchoscope. Objectives : To analyse the microscopic autofluorescence structure of normal and pathologic bronchial mucosae using FCFM during bronchoscopy. Methods : Bronchial FCFM and spectral analyses were performed at 488nm excitation wavelength on two bronchial specimens ex-vivo and in 29 individuals at high risk for lung cancer invivo. Biopsies of in vivo FCFM imaged areas were performed using autofluorescence bronchoscopy. Results : Ex-vivo and invivo microscopic and spectral analyses showed that the FCFM signal mainly originates from the elastin component of the basement membrane zone. Five distinct reproducible microscopic patterns were recognized in the normal areas from the trachea down to the more distal respiratory bronchi. In non previously biopsied areas of the proximal airways, one of these patterns was found in 30/30 normal epithelium, whereas alterations of the autofluorescence microstructure were observed in 19/22 metaplastic or dysplastic samples, 5/5 CIS and 2/2 invasive lesions. Disorganization of the fibered network could be found on 9/27 preinvasive lesions, compatible with early disruptions of the basement membrane zone. FCFM alterations were also observed in a tracheobronchomegaly syndrome and in a sarcoidosis case. Conclusions : Endoscopic FCFM represents a minimally invasive method to study specific basement membrane alterations associated with premalignant bronchial lesions in vivo. The technique may also be useful to study the bronchial wall remodelling in non malignant chronic bronchial diseases. www.clinicaltrials.gov NCT00213603 Key words: fluorescence, bronchoscopy, basement membrane, confocal microscopy, precancerous Conditions
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