Vibrational spectroscopic imaging and live cell video microscopy for studying differentiation of primary human alveolar epithelial cells.

Abstract:

: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.

journal_name

J Biophotonics

journal_title

Journal of biophotonics

authors

Vukosavljevic B,Hittinger M,Hachmeister H,Pilger C,Murgia X,Gepp MM,Gentile L,Huwer H,Schneider-Daum N,Huser T,Lehr CM,Windbergs M

doi

10.1002/jbio.201800052

subject

Has Abstract

pub_date

2019-06-01 00:00:00

pages

e201800052

issue

6

eissn

1864-063X

issn

1864-0648

journal_volume

12

pub_type

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