Abstract
We report a fiber optics-based intravital fluorescence imaging platform that includes epi-fluorescence microscopy and laser patterned-light stimulation system. The platform can perform real-time fluorescence imaging with a lateral resolution of ~4.9 μm while directly inserted into the intact mouse brain, optically stimulate vasoconstriction during real-time imaging, and avoid vessel damage in the penetration path of imaging probe. Using 473-nm patterned-light stimulation, we successfully modulated the vasoconstriction of a single targeted 37-μm-diameter blood vessel located more than 4.7 mm below the brain surface of a live SM22-ChR2 mouse. This platform may permit the hemodynamic studies associated with deeper brain neurovascular disorders.
Highlights
In neuroscience, it is important to understand the delivery of blood to cranial tissue and links between functional units, e.g., the functional activities of the neurovascular or gliovascular units, in vivo [1,2,3,4]
We have introduced an integrated intravital imaging platform including fiber optic-based epifluorescence microscopy and laser patterned-light stimulation systems
The platform can optically stimulate the vasoconstriction of cerebral vessels during real-time fluorescence imaging simultaneously
Summary
It is important to understand the delivery of blood to cranial tissue and links between functional units, e.g., the functional activities of the neurovascular or gliovascular units, in vivo [1,2,3,4]. Functional magnetic resonance imaging (fMRI) and positron emission tomography (PET) scans are the most commonly used imaging techniques exploiting hemodynamic responses to map brain functions [1, 6, 7] These imaging techniques have the disadvantage of difficulty in directly detecting intercellular responses, because they can only confirm the reaction to changes in blood flow caused by the simultaneous reaction of thousands of neurons [8]. Highresolution imaging optical systems such as conventional microscopy systems, including epifluorescence [9], confocal microscopy [10], and two-photon microscopy [11,12,13,14], have been applied to cerebrovascular studies These high-resolution imaging systems facilitate studies of the hemodynamics and blood cell–vessel wall interactions in the primary somatosensory cortex [11, 15], limb cortex, and parietal cortex [12, 14]. Because of the limited penetration of light into brain tissue, many researchers experience difficulties in studying deep-brain vascular-related diseases, such as Parkinson’s disease [2, 16]
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