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

Nature Methods volume 3, pages 99–108 (2006)Cite this article

Abstract

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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Figure 1: Experimental setup.
Figure 2: Hemorrhage via rupture of target vessel.
Figure 3: Extravasation of blood constituents into the parenchymal tissue without obstruction of flow in target vessel.
Figure 4: Intravascular clot in intact target arteriole.
Figure 5: Histological characterization of intravascular clots.
Figure 6: Changes in RBC velocity around subsurface intravascular clots.
Figure 7: Changes in RBC velocity around an intravascular clot with hemodilution.

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Acknowledgements

We thank Q. Cheng for assistance with the plasminogen activator experiments, E. Dolnick for assistance with the electronics, D. Pizzo and L. Thal for use of their photomicroscope, L. Schroeder and S. Siegel for discussions, and Coherent, Inc. for the loan of equipment. This work was funded by the David and Lucille Packard Foundation, the National Science Foundation (DBI/0455027), the National Institutes of Health (NS/041096, NS/043300, EB/003832 and RR/021907), a La Jolla Interfaces in Science Postdoctoral Fellowship to C.B.S. and a National Science Foundation Graduate Fellowship to N.N.

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Authors and Affiliations

  1. Department of Physics, 9500 Gilman Drive, University of California at San Diego, La Jolla, 92093, California, USA

    Nozomi Nishimura, Chris B Schaffer, Philbert S Tsai & David Kleinfeld

  2. Center for Theoretical Biological Physics, 9500 Gilman Drive, University of California at San Diego, La Jolla, 92093, California, USA

    Nozomi Nishimura, Chris B Schaffer, Philbert S Tsai & David Kleinfeld

  3. Department of Neurosciences, 9500 Gilman Drive, University of California at San Diego, La Jolla, 92093, California, USA

    Beth Friedman & Patrick D Lyden

  4. Graduate Program in Neurosciences, 9500 Gilman Drive, University of California at San Diego, La Jolla, 92093, California, USA

    Patrick D Lyden & David Kleinfeld

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Correspondence to David Kleinfeld.

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Nishimura, N., Schaffer, C., Friedman, B. et al. Targeted insult to subsurface cortical blood vessels using ultrashort laser pulses: three models of stroke. Nat Methods 3, 99–108 (2006). https://doi.org/10.1038/nmeth844

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