TY - JOUR
T1 - Vibrational spectroscopic imaging and live cell video microscopy for studying differentiation of primary human alveolar epithelial cells
AU - Vukosavljevic, Branko
AU - Hittinger, Marius
AU - Hachmeister, Henning
AU - Pilger, Christian
AU - Murgia, Xabier
AU - Gepp, Michael M.
AU - Gentile, Luca
AU - Huwer, Hanno
AU - Schneider-Daum, Nicole
AU - Huser, Thomas R
AU - Lehr, Claus Michael
AU - Windbergs, Maike
PY - 2019/6/1
Y1 - 2019/6/1
N2 - Alveolar type II (ATII) cells in the peripheral human lung spontaneously differentiate toward ATI cells, thus enabling air-blood barrier formation. Here, linear Raman and coherent anti-Stokes Raman scattering (CARS) microscopy are applied to study cell differentiation of freshly isolated ATII cells. The Raman spectra can successfully be correlated with gradual morphological and molecular changes during cell differentiation. Alveolar surfactant rich vesicles in ATII cells are identified based on phospholipid vibrations, while ATI-like cells are characterized by the absence of vesicular structures. Complementary, CARS microscopy allows for three-dimensional visualization of lipid vesicles within ATII cells and their secretion, while hyperspectral CARS enables the distinction between cellular proteins and lipids according to their vibrational signatures. This study paves the path for further label-free investigations of lung cells and the role of the pulmonary surfactant, thus also providing a basis for rational development of future lung therapeutics.
AB - Alveolar type II (ATII) cells in the peripheral human lung spontaneously differentiate toward ATI cells, thus enabling air-blood barrier formation. Here, linear Raman and coherent anti-Stokes Raman scattering (CARS) microscopy are applied to study cell differentiation of freshly isolated ATII cells. The Raman spectra can successfully be correlated with gradual morphological and molecular changes during cell differentiation. Alveolar surfactant rich vesicles in ATII cells are identified based on phospholipid vibrations, while ATI-like cells are characterized by the absence of vesicular structures. Complementary, CARS microscopy allows for three-dimensional visualization of lipid vesicles within ATII cells and their secretion, while hyperspectral CARS enables the distinction between cellular proteins and lipids according to their vibrational signatures. This study paves the path for further label-free investigations of lung cells and the role of the pulmonary surfactant, thus also providing a basis for rational development of future lung therapeutics.
KW - CARS microcopy
KW - cell imaging
KW - confocal laser scanning microscopy
KW - confocal Raman microscopy
KW - pneumocyte type II differentiation
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U2 - 10.1002/jbio.201800052
DO - 10.1002/jbio.201800052
M3 - Editorial
C2 - 30597770
AN - SCOPUS:85066856105
VL - 12
JO - Journal of Biophotonics
JF - Journal of Biophotonics
SN - 1864-063X
IS - 6
M1 - e201800052
ER -