| Autor: |
Yi-Chien Wu, Hyung-Geun Moon, Bindokas, Vytautas P., Phillips, Evan H., Gye Young Park, Lee Seung-Young, Steve |
| Předmět: |
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| Zdroj: |
American Journal of Respiratory Cell & Molecular Biology; Jul2023, Vol. 69 Issue 1, p13-E21, 30p |
| Abstrakt: |
Asthma is a chronic inflammatory airway disease driven by various infiltrating immune cell types into the lung. Optical microscopy has been used to study immune infiltrates in asthmatic lungs. Confocal laser scanning microscopy (CLSM) identifies the phenotypes and locations of individual immune cells in lung tissue sections by employing high-magnification objectives and multiplex immunofluorescence staining. In contrast, light-sheet fluorescence microscopy (LSFM) can visualize the macroscopic and mesoscopic architecture of wholemount lung tissues in three dimensions (3D) by adopting an optical tissue-clearing method. Despite each microscopy method producing image data with unique resolution from a tissue sample, CLSM and LSFM have not been applied together because of different tissue-preparation procedures. Here, we introduce a new approach combining LSFMand CLSM into a sequential imaging pipeline. We built a new optical tissue clearing workflow in which the immersion clearing agent can be switched from an organic solvent to an aqueous sugar solution for sequential 3D LSFMand CLSMof mouse lungs. This sequential combination microscopy offered quantitative 3D spatial analyses of the distribution of immune infiltrates in the samemouse asthmatic lung tissue at the organ, tissue, and cell levels. These results show that ourmethod facilitates multiresolution 3D fluorescencemicroscopy as a new imaging approach providing comprehensive spatial information for a better understanding of inflammatory lung diseases. [ABSTRACT FROM AUTHOR] |
| Databáze: |
Complementary Index |
| Externí odkaz: |
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