Autofluorescence multiphoton microscopy for visualization of tissue morphology and cellular dynamics in murine and human airways
Autor: | Gereon Hüttmann, Peter König, Holger Garn, Leigh M. Marsh, Sarah Kretschmer, Mario Pieper, Torsten Bölke, Barbara Wollenberg |
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Jazyk: | angličtina |
Rok vydání: | 2016 |
Předmět: |
0301 basic medicine
Male Pathology medicine.medical_specialty Ovalbumin Connective tissue Antigen-Presenting Cells Respiratory Mucosa Biology 01 natural sciences Pathology and Forensic Medicine 010309 optics Tissue Culture Techniques 03 medical and health sciences Tissue culture Mice Immune system Technical Report Cell Movement 0103 physical sciences medicine Animals Humans Antigen-presenting cell Molecular Biology Mice Inbred BALB C Lasers Optical Imaging Cell Biology Eosinophil respiratory system Epithelium Asthma Mice Inbred C57BL Trachea Autofluorescence Disease Models Animal Nasal Mucosa 030104 developmental biology medicine.anatomical_structure Microscopy Fluorescence Multiphoton Female Ex vivo Granulocytes |
Zdroj: | Laboratory Investigation; a Journal of Technical Methods and Pathology |
ISSN: | 1530-0307 0023-6837 |
Popis: | The basic understanding of inflammatory airway diseases greatly benefits from imaging the cellular dynamics of immune cells. Current imaging approaches focus on labeling specific cells to follow their dynamics but fail to visualize the surrounding tissue. To overcome this problem, we evaluated autofluorescence multiphoton microscopy for following the motion and interaction of cells in the airways in the context of tissue morphology. Freshly isolated murine tracheae from healthy mice and mice with experimental allergic airway inflammation were examined by autofluorescence multiphoton microscopy. In addition, fluorescently labeled ovalbumin and fluorophore-labeled antibodies were applied to visualize antigen uptake and to identify specific cell populations, respectively. The trachea in living mice was imaged to verify that the ex vivo preparation reflects the in vivo situation. Autofluorescence multiphoton microscopy was also tested to examine human tissue from patients in short-term tissue culture. Using autofluorescence, the epithelium, underlying cells, and fibers of the connective tissue, as well as blood vessels, were identified in isolated tracheae. Similar structures were visualized in living mice and in the human airway tissue. In explanted murine airways, mobile cells were localized within the tissue and we could follow their migration, interactions between individual cells, and their phagocytic activity. During allergic airway inflammation, increased number of eosinophil and neutrophil granulocytes were detected that moved within the connective tissue and immediately below the epithelium without damaging the epithelial cells or connective tissues. Contacts between granulocytes were transient lasting 3 min on average. Unexpectedly, prolonged interactions between granulocytes and antigen-uptaking cells were observed lasting for an average of 13 min. Our results indicate that autofluorescence-based imaging can detect previously unknown immune cell interactions in the airways. The method also holds the potential to be used during diagnostic procedures in humans if integrated into a bronchoscope. |
Databáze: | OpenAIRE |
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