Orthogonal‐plane fluorescence optical sectioning: Three‐dimensional imaging of macroscopic biological specimens

Abstract

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 guineapig cochlea. The lateral and axial resolutions of the images were 1 0 and 26 pm, respectively, within a 1.5-mm field of view.

An Electric Frequency-to-place Map for a Cochlear Implant Patient with Hearing in the Nonimplanted Ear

“…However, an estimate can be derived based on the 3D bony outline of the cochlea. Having a 3D volume of the patient_s cochlea, with the electrode array marked, also allows for predicting if the array is in scala tympani or scala vestibuli by comparing midmodiolar views with a 3D cochlear atlas generated from orthogonal-plane fluorescence optical sectioning microscopy (OPFOS) (Voie et al 1993). Unlike CT, OPFOS is capable of high-resolution imaging of the small tissue, as well as bony structures, of an excised cochlea.…”

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“…Light sheet microscopy or ultramicroscopy, originally developed in 1903 [41], reappeared in 1983 when it was applied to fixed and cleared tissue as orthogonal-plane fluorescence optical sectioning (OPFOS) [42], and as thin laser sheet imaging microscopy (TSLIM) in 2002 [43] finally being applied to whole fixed and cleared organs as ultramicroscopy [44]. However, it was not until Huisken et al in 2004 [45] presented selective plane illumination microscopy (SPIM) data applied to in vivo zebrafish imaging that light sheet microscopy really took off.…”

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