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. 2017 Dec 1;7(1):16761.
doi: 10.1038/s41598-017-17037-2.

Imaging viscosity of intragranular mucin matrix in cystic fibrosis cells

Sebastian Requena  1 , Olga Ponomarchuk  2   3 , Marlius Castillo  4 , Jonathan Rebik  4 , Emmanuelle Brochiero  2   5 , Julian Borejdo  1 , Ignacy Gryczynski  1 , Sergei V Dzyuba  6 , Zygmunt Gryczynski  1   7 , Ryszard Grygorczyk  8   9 , Rafal Fudala  10
Affiliations

Imaging viscosity of intragranular mucin matrix in cystic fibrosis cells

Sebastian Requena et al. Sci Rep. .

Abstract

Abnormalities of mucus viscosity play a critical role in the pathogenesis of several respiratory diseases, including cystic fibrosis. Currently, there are no approaches to assess the rheological properties of mucin granule matrices in live cells. This is the first example of the use of a molecular rotor, a BODIPY dye, to quantitatively visualize the viscosity of intragranular mucin matrices in a large population of individual granules in differentiated primary bronchial epithelial cells using fluorescence lifetime imaging microscopy.

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Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Structures of the dyes and FLIM images. FLIM images of the BODIPY rotor and non-rotor dyes incubated with CF (a and c) and non-CF (b and d) cells. Images scales are 80 × 80 µm. Insets: examples of high-resolution scans of the highlighted regions of interest.
Figure 2
Figure 2
Viscosity of intragranular mucin matrix. The viscosity distribution of intragranular mucin matrix in CF (a and c) and non-CF (b and d) cells using rotor and non-rotor BODIPY dyes. The analysis was performed in three independent experiments with a total of 10–12 images analyzed per set. The number of granules (N) analyzed in each case is given at the right top corner of each panel. Blue, green and red lines are fits: blue – overall fits, green and red lines are fits for lower and higher viscosity populations.
See this image and copyright information in PMC

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