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Visualizing Cytoplasmic Flow During Single-cell Wound Healing in Stentor coeruleus
Published on: December 19, 2013
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Label-free concurrent 5-modal microscopy (Co5M) resolves unknown spatio-temporal processes in wound healing
Markus Seeger1,2, Christoph Dehner1,2, Dominik Jüstel1,2,3
1Chair of Biological Imaging, Central Institute for Translational Cancer Research (TranslaTUM), School of Medicine, Technical University of Munich, Munich, Germany.
Communications Biology
|September 7, 2021
Summary
Concurrent 5-modal microscopy (Co5M) enables label-free, in vivo visualization of biological processes. This advanced technique captures multiple contrasts simultaneously, offering unprecedented insights into complex physiological and pathological changes without disturbing the biological system.
Area of Science:
- * Biomedical imaging
- * In vivo microscopy
- * Label-free imaging
Background:
- * Non-invasive investigation of biological processes is challenging due to the need for multiple contrast mechanisms.
- * Intravital microscopy typically requires transgenic labels, which can disturb biological systems.
- * Existing methods are limited in simultaneously capturing diverse contrast mechanisms for comprehensive analysis.
Purpose of the Study:
- * To introduce concurrent 5-modal microscopy (Co5M) for label-free in vivo observation.
- * To demonstrate Co5M's capability in simultaneously capturing optoacoustic, two-photon excitation fluorescence, second and third harmonic generation, and brightfield contrasts.
- * To apply Co5M for non-invasive visualization and quantitative monitoring of wound healing biomarkers.
Main Methods:
- * Development and application of concurrent 5-modal microscopy (Co5M).
- * Simultaneous capture of five contrast mechanisms: optoacoustic, two-photon excitation fluorescence, second harmonic generation, third harmonic generation, and brightfield.
- * Label-free in vivo imaging applied to wound healing models.
Main Results:
- * Co5M successfully visualized multiple wound healing biomarkers non-invasively.
- * Quantitative monitoring of wound shape, microvascular and collagen density, vessel size and fractality, and sebaceous gland plasticity was achieved.
- * Detailed insights into the interplay of wound closure, vasodilation, angiogenesis, skin contracture, and epithelial reformation were obtained.
- * Analysis revealed dynamic spatial and temporal changes inaccessible by other methods.
Conclusions:
- * Co5M provides a novel platform for label-free, multi-contrast in vivo imaging.
- * The technique offers unique insights into complex biological processes, such as wound healing.
- * Co5M challenges conventional biological observation by enabling simultaneous acquisition of multiple pathophysiological parameters without labels.
