Targeted Occlusion of Individual Pial Vessels of Mouse Cortex

Zachary Taylor, Andy Shih
2013 Bio-protocol  
Targeted photothrombosis is a method to occlude individual arterioles and venules that lie on the surface of the cerebral cortex. It has been used to study collateral flow patterns within the pial vascular network following occlusion of single surface vessels (Schaffer et al., 2006; Nguyen et al., 2011) , as well as to generate localized ischemic strokes following occlusion of single penetrating vessels (Nishimura et al., 2007; Drew et al., 2010; Shih et al., 2013) . The intravascular clot is
more » ... rmed by irradiation of a target vessel with a focused green laser after injection of a circulating photosensitizing agent, Rose Bengal (Watson et al., 1985) . We briefly describe modifications of custom-designed and commercial two-photon imaging systems required to introduce a green laser for photothrombosis. We further provide instructions on how to occlude a single penetrating arteriole within the somatosensory cortex of an anesthetized mouse. Materials and Reagents Mouse with cranial window implant 2. Buprenorphine hydrochloride (Buprenex  ) (Butler Schein, catalog number: 031919) 3. Isoflurane (Butler Schein, catalog number: 029405) 4. Ophthalmic ointment (Butler Schein, catalog number: 039886) 5. Cove glass (no. 0 thickness) (Thomas Scientific, catalog number: 6661B40) 6. Filter paper (Thermo Fisher Scientific, catalog number: S47573B) 7. 60 mm culture dish (Thermo Fisher Scientific, catalog number: 130181) 8. Distilled water 9. Glucose (Sigma-Aldrich, catalog number: G8270) 10. HEPES (Sigma-Aldrich, catalog number: H7006) 11. Artificial cerebral spinal fluid (ACSF) (see Recipes) 12. FITC-dextran (Sigma-Aldrich, catalog number: FD2000S) solution (see Recipes) 13. Rose Bengal (Sigma-Aldrich, catalog number: 632-69-9) solution (see Recipes)
doi:10.21769/bioprotoc.897 fatcat:axfz535lqrhbbk7jdnrpu5inky