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Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke

Nozomi Nishimura, Chris B. Schaffer, Beth Friedman, Philbert S. Tsai, Patrick D. Lyden, and David Kleinfeld

Nature Methods (2006)

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We present a method to produce vascular disruptions within rat brain parenchyma that targets single microvessels. We used two-photon microscopy to image vascular architecture, to select a vessel for injury and to measure blood-flow dynamics. We irradiated the vessel with high-fluence, ultrashort laser pulses and achieved three forms of vascular insult. (i) Vessel rupture was induced at the highest optical energies; this provides a model for hemorrhage. (ii) Extravasation of blood components was induced near the lowest energies and was accompanied by maintained flow in the target vessel. (iii) An intravascular clot evolved when an extravasated vessel was further irradiated. Such clots dramatically impaired blood flow in downstream vessels, in which speeds dropped to as low as ~10% of baseline values. This demonstrates that a single blockage to a microvessel can lead to local cortical ischemia. Lastly, we show that hemodilution leads to a restoration of flow in secondary downstream vessels.

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Two-photon imaging of cortical surface microvessels reveals a robust redistribution of blood flow after vascular occlusion

Chris B. Schaffer, Beth Friedman, Nozomi Nishimura, Lee F. Schroeder, Philbert S. Tsai, Ford F. Ebner, Patrick D. Lyden, and David Kleinfeld

Public Library of Science Biology (2006)

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A highly interconnected network of arterioles overlies mammalian cortex to route blood to the cortical mantle. Here we test if this angioarchitecture can ensure that the supply of blood is redistributed after vascular occlusion. We use rodent parietal cortex as a model system and image the flow of red blood cells in individual microvessels. Changes in flow are quantified in response to photothrombotic occlusions to individual pial arterioles as well as to physical occlusions of the middle cerebral artery (MCA), the primary source of blood to this network. We observe that perfusion is rapidly reestablished at the first branch downstream from a photothrombotic occlusion through a reversal in flow in one vessel. More distal downstream arterioles also show reversals in flow. Further, occlusion of the MCA leads to reversals in flow through approximately half of the downstream but distant arterioles. Thus the cortical arteriolar network supports collateral flow that may mitigate the effects of vessel obstruction, as may occur secondary to neurovascular pathology.

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Minimally disruptive laser-induced breakdown in water

Eli N. Glezer, Chris B. Schaffer, Nozomi Nishimura, and Eric Mazur

Optics Letters (1997)

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We produce minimally disruptive breakdown in water by using tightly focused 100-fs laser pulses and demonstrate the potential use of this technique in microsurgery of the eye. Using time-resolved imaging and piezoelectric pressure detection, we measure the magnitude and speed of propagation of the pressure wave produced in the breakdown. Compared with breakdown with longer pulses, here there is a much lower energy threshold for breakdown of 0.2 mJ , a smaller shock zone diameter (11 mm for 1-mJ pulses), and consistent energy deposition.

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