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Intravital two-photon imaging: a versatile tool for dissecting the immune system
1Laboratory of Biological Imaging, WPI-Immunology Frontier Research Center, Osaka University, Osaka 565-0871, Japan.
Insights
Two-photon microscopy allows real-time visualization of immune cell dynamics deep within living tissues. This advanced imaging technique reveals the intricate, dynamic nature of the immune system, crucial for understanding physiology and pathology.
Area of Science:
- Immunology
- Microscopy
- Cell Biology
Background:
- The immune system is highly dynamic, with cells constantly circulating and interacting.
- Conventional methods offer limited insight into immune cell dynamics in vivo.
- Two-photon microscopy has emerged as a revolutionary tool for biological imaging.
Purpose of the Study:
- To review recent findings in intravital two-photon imaging of the immune system.
- To highlight the capabilities of two-photon microscopy in studying immune cell behavior.
- To emphasize the importance of real-time imaging for understanding immune system function.
Main Methods:
- Utilizing near-infrared excitation lasers for deep tissue penetration.
- Minimizing photobleaching and toxicity for extended live imaging.
- Applying two-photon excitation-based laser microscopy for intravital imaging.
Main Results:
- Two-photon microscopy enables visualization of living cell behavior deep within tissues.
- It allows for extended observation of live, intact specimens.
- Recent findings showcase its power in dissecting immune system dynamics.
Conclusions:
- Intravital two-photon imaging is a powerful tool for dissecting immune system mechanisms.
- It overcomes limitations of conventional methods in studying immune cell dynamics.
- This technology revolutionizes the study of dynamic cellular processes in vivo.
Abstract:
During the past decade, multi-photon or 'two-photon' excitation microscopy has launched a new era in the field of biological imaging. The near-infrared excitation laser for two-photon microscopy can penetrate thicker specimens, enabling the visualisation of living cell behaviour deep within tissues and organs without thin sectioning. The minimised photobleaching and toxicity enables the visualisation of live and intact specimens for extended periods. In this brief review, recent findings in intravital two-photon imaging for the physiology and pathology of the immune system are discussed. The immune system configures highly dynamic networks, where many cell types actively travel throughout the body and interact with each other in specific areas. Hence, real-time intravital imaging may be a powerful tool for dissecting the mechanisms of this dynamic system. The most unique characteristic of the immune system is its highly dynamic nature. A variety of cell types, such as lymphocytes, macrophages and dendritic cells (DCs), are continuously circulating throughout the body, migrating through the peripheral tissues and interacting with each other in their respective niches. Conventional methodologies in immunology, such as flow cytometry, cell or tissue culture, biochemistry and histology, have brought tremendous achievement within this field, although the dynamics of immune cells in an entire animal remain less clear. Technological progress of fluorescence microscopy has enabled us to visualise the intact biological phenomenon that has been uninvestigated. Among the advancements, the recent emergence and prevalence of two-photon, excitation-based, laser microscopy has revolutionised the research field, such that the dynamic behaviour of cells deep inside living organs can be visualised and analysed.
