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@inproceedings{Patel:2015,
author = {Patel, BV and Tatham, KC and Wilson, MR and O'Dea, K and Takata, M},
pages = {A3926--A3926},
publisher = {ATS Journals},
title = {In Vivo Compartmental Labeling of the Mouse Lung},
year = {2015}
}
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TY - CPAPER
AB - ShareModerators: G.N. Maksym, PhD, R.S. Harris, MD, J.C. Sieren, PhDSession Info: Mini Symposium, [C18] STATE OF PLAY IN RESPIRATORY STRUCTURE AND FUNCTION: SEEING IS BELIEVINGDay/Date: Tuesday, May 19, 2015Session Time: 9:30 AM - 11:30 AMRoom: Mile High Ballroom 2C/3C (Lower Level)Location: Colorado Convention CenterIn Vivo Compartmental Labeling of the Mouse Lung, [Publication Page: A3926]B.V. Patel, MBBS MRCP FRCA PhD, K.C. Tatham, MBBS, M.R. Wilson, PhD, K.P. O'Dea, PhD, M. Takata, MDLondon/UKRationaleCurrent methods for compartmental analyses of lungs using bronchoalveolar lavage (BAL) and/or lung perfusion have significant limitations for accurate determination of location and phenotype of lung leukocytes. BAL retrieves only <25% of intra-alveolar cells (1) and lung perfusion fails to remove vascular marginated cell populations (2). We present a novel method of in vivo antibody labelling to facilitate the accurate compartmental investigation of mouse lung leukocytes by flow cytometry.MethodsAnesthetized C57BL6 mice underwent tracheostomy and venous cannulation. An anti-CD45 antibody (PE-conjugated) was injected intravenously and allowed to circulate for 5 minutes. Mice were then exsanguinated and lungs immediately flooded intratracheally with another anti-CD45 antibody (PE-Cy5-conjugated). Lungs were harvested, and single cell suspensions prepared for flow cytometric analysis using a panel of myeloid markers. This in vivo labelling protocol was also applied to mice exposed to acid-aspiration induced lung injury (3). To confirm the separation between the vascular and interstitial cell populations, a group of mice underwent intravenous labelling alone followed by lung perfusion for 15 minutes using an isolated-perfused lung apparatus (2).ResultsThe combined in vivo intravenous and intratracheal CD45 labelling enabled robust separation of the alveolar, interstitial, and vascular compartments of the lung by flow cytometry, with <0.3% of leukocytes staining
AU - Patel,BV
AU - Tatham,KC
AU - Wilson,MR
AU - O'Dea,K
AU - Takata,M
EP - 3926
PB - ATS Journals
PY - 2015///
SN - 1073-449X
SP - 3926
TI - In Vivo Compartmental Labeling of the Mouse Lung
ER -