In Vivo Two Photon Imaging
In Vivo Two Photon Imaging - Related Videos
04:32
Two-Photon In Vivo Imaging of the Mouse Retina
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2025
Source: Wang, Z., et al. Transpupillary Two-Photon In Vivo Imaging of the Mouse Retina. J. Vis. Exp. (2021).This video demonstrates a technique for in vivo retinal imaging using two-photon microscopy. Anesthetized mice are prepared for imaging by securing their heads, Applying eye lubricant, and placing a coverslip over the eye. Retinal ganglion cells (RGCs) expressing fluorescent proteins are first visualized using epifluorescence for alignment. Finally, two-photon imaging is employed,...
10:24
In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy
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Cited by 42 •
2012
A minimally invasive protocol to stabilize the mouse spinal column and perform repetitive in vivo spinal cord imaging using two-photon microscopy is described. This method combines a spinal stabilization device and an anesthetic regimen to minimize respiratory-induced movements and produce raw imaging data that require no alignment or other post-processing.
02:58
Two-Photon Microscopy for In Vivo Imaging of Mouse Brain Microvasculature
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2025
Source: Nishimura, C. et al. Quantification of Cerebral Vascular Architecture using Two-photon Microscopy in a Mouse Model of HIV-induced Neuroinflammation. J. Vis. Exp. (2016). This video demonstrates the in vivo visualization of cortical capillaries in the mouse brain using two-photon microscopy. It outlines the steps for injecting a fluorescent dye into an anesthetized mouse with a thin-skull cortical window, preparing the setup for imaging, and acquiring detailed images of brain...
02:42
In Vivo Two-Photon Imaging of Microglial Dynamics in the Mouse Hippocampus
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2025
Source: Kamei, R., et. al., In Vivo Chronic Two-Photon Imaging of Microglia in the Mouse Hippocampus. J. Vis. Exp. (2022)The video demonstrates the setup and imaging process for in vivo two-photon microscopy of microglial dynamics in the hippocampal CA1 (Cornu ammonis 1) and dentate gyrus, ensuring optimal optical alignment and structural integrity assessment.
10:07
In Vivo Two-photon Imaging Of Experience-dependent Molecular Changes In Cortical Neurons
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Cited by 31 •
2013
Experience-dependent molecular changes in neurons are essential for the brain's ability to adapt in response to behavioral challenges. An in vivo two-photon imaging method is described here that allows the tracking of such molecular changes in individual cortical neurons through genetically encoded reporters.
FAQs
Brief near-infrared laser pulses generate fluorescence primarily at the microscope’s focal point, where two-photon excitation occurs. Regions above and below that plane receive less excitation and therefore contribute less out-of-focus fluorescence. This focal restriction helps distinguish structures within complex tissue and supports more precise visualization of cellular events in their native biological environment.
Near-infrared excitation helps the microscope collect information deeper within scattering tissue than would be practical with more surface-limited imaging approaches. Combined with focal excitation, it allows researchers to examine structures beneath the tissue surface while reducing unwanted fluorescence from regions outside the focal plane. This capability is important when intact tissue must remain present during observation.
Restricting excitation to the focal point limits unnecessary light exposure in surrounding tissue, reducing out-of-focus photodamage during imaging. That feature supports repeated observation of the same living tissue, allowing researchers to follow changing cellular behavior over time rather than relying only on separate samples collected at different stages.
Researchers can return to living tissue for repeated imaging sessions and compare cellular behavior across observations. This longitudinal approach links movement, activity, and interactions with changes in tissue function or disease progression. The resulting observations emphasize dynamics, such as how immune cells migrate or how cellular relationships change within an intact biological environment.
In biology, the method can visualize neuronal activity, blood flow, immune-cell migration, and interactions between cells. Observing these processes in intact tissue preserves the surrounding environment that influences cellular behavior. Consequently, researchers can relate individual cellular events to broader tissue function instead of examining isolated cells without their normal biological context.
Its ability to support repeated imaging makes it possible to associate changing cellular behavior with the progression of disease-related tissue changes. Researchers can examine how neuronal activity, blood flow, immune-cell migration, or cellular interactions evolve in living tissue. This temporal perspective provides biological context that a single observation cannot capture and helps connect cell-level dynamics with tissue function.