RESUMO
This paper describes a methodology for three-dimensional (3D) computer-aided reconstruction of the guinea pig cochlea using orthogonal-plane fluorescence optical sectioning. Specimens are sectioned optically, allowing them to remain intact during observation. Equations to correct the data for specimen translation and rotation are developed and 3D reconstructions of the scala tympani, round window membrane, and cochlear aqueduct are presented. The error associated with the reconstruction is estimated to be < 19 microns.
Assuntos
Cóclea/anatomia & histologia , Processamento de Imagem Assistida por Computador , Animais , Aqueduto da Cóclea/anatomia & histologia , Terminais de Computador , Desenho Assistido por Computador , Cobaias , Aumento da Imagem , Processamento de Imagem Assistida por Computador/instrumentação , Processamento de Imagem Assistida por Computador/métodos , Armazenamento e Recuperação da Informação , Microscopia de Fluorescência , Microtomia , Rotação , Janela da Cóclea/anatomia & histologia , Rampa do Tímpano/anatomia & histologiaRESUMO
An imaging technique called orthogonal-plane fluorescence optical sectioning (OPFOS) was developed to image the internal architecture of the cochlea. Expressions for the three-dimensional point spread function and the axial and lateral resolution are derived. Methodologies for tissue preparation and for construction, alignment, calibration and characterization of an OPFOS apparatus are presented. The instrument described produced focused, high-resolution images of optical sections of an intact, excised guinea-pig cochlea. The lateral and axial resolutions of the images were 10 and 26 microns, respectively, within a 1.5-mm field of view.