title: Retinal regeneration following OCT-guided laser injury in zebrafish.
abstract: OBJECTIVE
  Establish a focal injury/regeneration model in zebrafish using laser photocoagulation guided by optical coherence tomography (OCT).
  
  
  METHODS
  Adult zebrafish were imaged by OCT and confocal scanning laser ophthalmoscopy (cSLO) in room air through a contact lens. Using a beam combiner, 532-nm laser photocoagulation was applied using the OCT C-scan image for targeting. Laser spots of 42 to 47 mW were delivered to the retina. At multiple intervals post injury, fish were imaged using both OCT and cSLO to follow the progression of each lesion. Histologic sections and TUNEL staining were performed to monitor the injury response.
  
  
  RESULTS
  Round lesions (26057 ± 621 μm(2)) localized to the outer retina were successfully applied. Laser application was visualized by real-time OCT and lesions were detectable by both OCT and cSLO in vivo. Lesion size increased 1 day post lesion then decreased in size. Histologic sections showed focal areas of damage localized primarily to the outer retina. By 3 weeks, the damaged areas had regenerated and a fully laminated structure was re-established. However, subtle changes can still be detected by OCT, cSLO imaging, and histology. Infrared darkfield imaging was more sensitive than OCT at revealing subtle changes in regenerated areas.
  
  
  CONCLUSIONS
  Optical coherence tomography-guided laser photocoagulation is a useful tool for inducing localized lesions and studying retinal regeneration in zebrafish. This novel method will allow us to characterize the cellular and molecular changes that take place at the interface between normal and damaged tissue. Regeneration can be observed using high-resolution OCT and cSLO imaging in vivo.
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio, United States.
authors.forename: Rose M
authors.initials: RM
authors.lastname: DiCicco
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio, United States.
authors.forename: Brent A
authors.initials: BA
authors.lastname: Bell
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio, United States.
authors.forename: Charles
authors.initials: C
authors.lastname: Kaul
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio, United States.
authors.forename: Joe G
authors.initials: JG
authors.lastname: Hollyfield
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio, United States.
authors.forename: Bela
authors.initials: B
authors.lastname: Anand-Apte
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio, United States.
authors.forename: Brian D
authors.initials: BD
authors.lastname: Perkins
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio, United States.
authors.forename: Yuankai K
authors.initials: YK
authors.lastname: Tao
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, Ohio, United States.
authors.forename: Alex
authors.initials: A
authors.lastname: Yuan
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D005500:Follow-Up Studies; D017075:Laser Coagulation / Q000009:adverse effects; D009887:Ophthalmoscopy; D012160:Retina / Q000293:injuries / Q000473:pathology*; D012162:Retinal Degeneration / Q000209:etiology / Q000473:pathology*; D041623:Tomography, Optical Coherence / Q000379:methods*; D015027:Zebrafish
keywords: laser; optical coherence tomography; regeneration; retina; scanning laser ophthalmoscopy