Multiphoton Intravital Imaging
Multiphoton Intravital Imaging - Related Videos
06:29
Inducing Ischemia-reperfusion Injury in the Mouse Ear Skin for Intravital Multiphoton Imaging of Immune Responses
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Cited by 15 •
2016
This protocol describes the induction of an ischemia-reperfusion (IR) model on mouse ear skin using magnet clamping. Using a custom-built intravital imaging model, we study in vivo inflammatory responses post-reperfusion. The rationale behind the development of this technique is to extend the understanding of how leukocytes respond to skin IR injury.
11:44
Long Term Intravital Multiphoton Microscopy Imaging of Immune Cells in Healthy and Diseased Liver Using CXCR6.Gfp Reporter Mice
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Cited by 27 •
2015
Stable intravital high-resolution imaging of immune cells in the liver is challenging. Here we provide a highly sensitive and reliable method to study migration and cell-cell-interactions of immune cells in mouse liver over long periods (about 6 hours) by intravital multiphoton laser scanning microscopy in combination with intensive care monitoring.
08:24
Extended Time-lapse Intravital Imaging of Real-time Multicellular Dynamics in the Tumor Microenvironment
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Cited by 25 •
2016
This protocol describes the use of multiphoton microscopy to perform extended time-lapse imaging of multicellular interactions in real time, in vivo at single cell resolution.
07:50
Multiphoton Intravital Imaging for Monitoring Leukocyte Recruitment during Arteriogenesis in a Murine Hindlimb Model
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Cited by 1 •
2021
The recruitment of leukocytes and platelets constitutes an essential component necessary for the effective growth of collateral arteries during arteriogenesis. Multiphoton microscopy is an efficient tool for tracking cell dynamics with high spatio-temporal resolution in vivo and less photo-toxicity to study leukocyte recruitment and extravasation during arteriogenesis.
08:41
Intravital Imaging of the Mouse Popliteal Lymph Node
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Cited by 27 •
2012
Recent advances in 2-photon microscopy have enabled real-time in situ imaging of live tissues in animal models, thereby enhancing our ability to investigate cellular behavior in both physiologic and pathologic conditions. Here, we outline the preparations required to perform intravital imaging of the mouse popliteal lymph node.
FAQs
Near-infrared light supports deeper fluorescence imaging in tissue, while short pulses concentrate excitation at the selected focal plane. Because fluorescence is generated primarily where the laser is focused, regions above and below that plane contribute less unwanted signal. This focal restriction helps preserve image contrast when investigators examine cellular behavior within living tissue.
The focal plane controls where two-photon or multiphoton excitation takes place. Light outside that location does not produce the same localized excitation, which limits unnecessary exposure of surrounding tissue. Reduced out-of-focus photodamage helps investigators observe dynamic biological events while minimizing disruption from the imaging process itself, an important consideration when studying living systems.
Tissue can scatter emitted or transmitted light, which may reduce the clarity of fluorescence signals from deeper regions. Multiphoton intravital imaging addresses this challenge by combining near-infrared excitation with focal-plane fluorescence generation. The resulting reduction in unwanted scattered signal supports clearer visualization of cellular and tissue dynamics within the physiological environment.
Investigators observe a selected tissue region in a living organism and use focal-plane excitation to visualize fluorescence as biological events unfold. Rather than examining disease only after tissue removal, the approach reveals cellular behavior in its physiological context. This makes it possible to relate changing cell activity and tissue conditions to disease development over time.
Applications include tracking immune-cell migration, monitoring tumor progression, observing blood-flow changes, and examining drug distribution. These processes occur dynamically, so real-time imaging can show how cells and tissues behave rather than providing only a static endpoint. The resulting observations help connect local biological activity with broader patterns of disease progression.
By visualizing drug distribution and tissue behavior in real time, the method can show how treatment-related changes correspond with cellular and disease processes. Researchers can compare observed molecular or cellular behavior with progression or response in the living tissue environment. This connection supports more informative preclinical research before treatment effects are evaluated in later studies.