Abstract
Multiphoton imaging is a promising approach for addressing current issues in systems biology and high-content investigation of embryonic development. Recent advances in multiphoton microscopy, including light-sheet illumination, optimized laser scanning, adaptive and label-free strategies, open new opportunities for embryo imaging. However, the literature is often unclear about which microscopy technique is most adapted for achieving specific experimental goals. In this review, we describe and discuss the key concepts of imaging speed, imaging depth, photodamage, and nonlinear contrast mechanisms in the context of recent advances in live embryo imaging. We illustrate the potentials of these new imaging approaches with a selection of recent applications in developmental biology.
🔬 Techniques
🧬 Organisms
✨ Fluorophores
🧪 Sample Preparation
🏭 Microscope Brands
📷 Detectors
🏛️ Research Organizations (ROR)
Affiliated research institutions:
📊 Figures
Figure 1
Strategies for improving acquisition speed in current fluorescence microscopy techniques
Similar strategies have been developed in linear (a-c) and nonlinear (d-f) microscopy. Both point-scanning (a and d) and multifocal (b and e) approaches use a collinear geometry: the illumination and ...
Figure 2
Selected applications of advanced multiphoton microscopy in developmental biology
This figure illustrates various recent applications of multiphoton microscopy in developmental biology. (a) Long term imaging of Drosophila embryos using light-sheet 2p-microscopy (reprinted from [ 16...
Figure 3
Imaging depth in point-scanning and light-sheet microscopy: comparison of 1-photon and 2-photon excited fluorescence
The principles governing imaging depth are analyzed in three steps: sample illumination (a-d), fluorescence excitation (e-h) and fluorescent detection (i-l). 2p-microscopy uses longer excitation wavel...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
💬 Discussion
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