Related Experiment Video
Updated: Jul 16, 2025
Long-term Intravital Immunofluorescence Imaging of Tissue Matrix Components with Epifluorescence and Two-photon Microscopy
Published on: April 22, 2014
Dynamic Multiplex Tissue Imaging in Inflammation Research
Stefan Uderhardt1,2,3, Georgiana Neag1,2,3, Ronald N Germain4,5
1Department of Medicine 3-Rheumatology and Immunology, Friedrich-Alexander-University Erlangen-Nürnberg and Universitätsklinikum Erlangen, Erlangen, Germany.
Insights
Two-photon microscopy (2PM) enables in vivo imaging of deep tissues, revealing dynamic immune cell behaviors crucial for understanding inflammation. This technology helps correlate cellular dynamics with in situ function and molecular patterns within complex tissue environments.
Area of Science:
- Immunology
- Cell Biology
- Biomedical Imaging
Background:
- Inflammation involves dynamic immune cell interactions within complex tissue environments.
- Assessing these cellular dynamics in vivo is crucial due to limitations of ex vivo models.
- Deep tissue imaging is necessary to observe these processes far below organ surfaces.
Purpose of the Study:
- To illustrate how two-photon microscopy (2PM) facilitates in vivo studies of immune cell dynamics.
- To connect observed immune cell behaviors to their in situ functions and governing molecular patterns.
- To review practical aspects and potential confounding factors of 2PM in biological research.
Main Methods:
- Utilizing two-photon excitation lasers integrated with existing imaging systems.
- Performing long-term, deep-tissue imaging in vivo.
- Analyzing studies that apply 2PM to investigate immune cell behavior in inflammatory contexts.
Main Results:
- 2PM allows visualization of immune cell migration and interactions deep within tissues.
- Studies demonstrate the link between immune cell dynamics and their functional roles in situ.
- Molecular patterns influencing immune cell behavior in vivo have been elucidated using 2PM.
Conclusions:
- Two-photon microscopy is a powerful tool for studying dynamic immune cell behavior in vivo.
- Understanding in situ cell dynamics is essential for comprehending inflammatory processes.
- Careful consideration of 2PM methodologies is needed to ensure reliable scientific conclusions.
Abstract:
Inflammation is a highly dynamic process with immune cells that continuously interact with each other and parenchymal components as they migrate through tissue. The dynamic cellular responses and interaction patterns are a function of the complex tissue environment that cannot be fully reconstructed ex vivo, making it necessary to assess cell dynamics and changing spatial patterning in vivo. These dynamics often play out deep within tissues, requiring the optical focus to be placed far below the surface of an opaque organ. With the emergence of commercially available two-photon excitation lasers that can be combined with existing imaging systems, new avenues for imaging deep tissues over long periods of time have become available. We discuss a selected subset of studies illustrating how two-photon microscopy (2PM) has helped to relate the dynamics of immune cells to their in situ function and to understand the molecular patterns that govern their behavior in vivo. We also review some key practical aspects of 2PM methods and point out issues that can confound the results, so that readers can better evaluate the reliability of conclusions drawn using this technology.
More Related Videos
11:00Visualization, Quantification, and Mapping of Immune Cell Populations in the Tumor Microenvironment
Published on: March 25, 2020
06:05Author Spotlight: Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment
Published on: June 2, 2023