title: Rose Bengal Photothrombosis by Confocal Optical Imaging In Vivo: A Model of Single Vessel Stroke.
abstract: In vivo imaging techniques have increased in utilization due to recent advances in imaging dyes and optical technologies, allowing for the ability to image cellular events in an intact animal. Additionally, the ability to induce physiological disease states such as stroke in vivo increases its utility. The technique described herein allows for physiological assessment of cellular responses within the CNS following a stroke and can be adapted for other pathological conditions being studied. The technique presented uses laser excitation of the photosensitive dye Rose Bengal in vivo to induce a focal ischemic event in a single blood vessel. The video protocol demonstrates the preparation of a thin-skulled cranial window over the somatosensory cortex in a mouse for the induction of a Rose Bengal photothrombotic event keeping injury to the underlying dura matter and brain at a minimum. Surgical preparation is initially performed under a dissecting microscope with a custom-made surgical/imaging platform, which is then transferred to a confocal microscope equipped with an inverted objective adaptor. Representative images acquired utilizing this protocol are presented as well as time-lapse sequences of stroke induction. This technique is powerful in that the same area can be imaged repeatedly on subsequent days facilitating longitudinal in vivo studies of pathological processes following stroke.
authors.affiliation: Department of Cellular and Structural Biology, The University of Texas Health Science Center San Antonio.
authors.forename: Lora
authors.initials: L
authors.lastname: Talley Watts
authors.affiliation: Department of Cellular and Structural Biology, The University of Texas Health Science Center San Antonio.
authors.forename: Wei
authors.initials: W
authors.lastname: Zheng
authors.affiliation: School of Medicine, The University of Texas Health Science Center San Antonio.
authors.forename: R Justin
authors.initials: RJ
authors.lastname: Garling
authors.affiliation: Cell & Tissue Imaging Center, St. Jude Childrens Research Hospital.
authors.forename: Victoria C
authors.initials: VC
authors.lastname: Frohlich
authors.affiliation: Department of Cellular and Structural Biology, The University of Texas Health Science Center San Antonio; lechleiter@uthscsa.edu.
authors.forename: James Donald
authors.initials: JD
authors.lastname: Lechleiter
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D007511:Ischemia / Q000209:etiology / Q000473:pathology; D007834:Lasers; D051379:Mice; D012395:Rose Bengal; D013927:Thrombosis / Q000209:etiology* / Q000473:pathology; D041623:Tomography, Optical Coherence / Q000379:methods*
chemical_list: D012395:Rose Bengal