Light sheet microscopy of the gerbil cochlea

Abstract

Light sheet fluorescence microscopy (LSFM) provides a rapid and complete threedimensional image of the cochlea. The method retains anatomical relationships-on a micrometer scale-between internal structures such as hair cells, basilar membrane (BM), and modiolus with external surface structures such as the round and oval win

In Situ 3D-Imaging of the Inner Ear Synapses with a Cochlear Implant

“…Since the working distance of our microscope is the main limiting factor for imaging of the cochlea synapses, other imaging techniques may be applicable. One option for whole cochlea imaging is light sheet fluorescence microscopy (LSFM) [ 38 , 41 , 42 ] which also requires optical clearing of the tissue. This method as well offers the opinion to leave an implant in situ to evaluate both the amount of fibrosis and the volumetric degree of hair cell loss [ 38 ].…”

“…Nevertheless conventional LSFM reaches its limitations by visualizing the cellular level. Hutson et al had to rehydrate and dissect the cochleae for visualization of the subcellular level with CLSM after LSFM, increasing the work load due to additional tissue preparation [ 42 , 44 ]. Meanwhile a new method of LSFM is developed called cleared tissue axially swept light-sheet microscopy (ctASLM) [ 45 ], which achieves subcellular resolution in cleared tissues.…”

Evaluation of the Efficacy of Dexamethasone-Eluting Electrode Array on the Post-Implant Cochlear Fibrotic Reaction by Three-Dimensional Immunofluorescence Analysis in Mongolian Gerbil Cochlea

“…Light-sheet microscopy is a technique of choice for studying biological structures in three dimensions. It allows three-dimensional analysis of whole cochlear samples without dissection trauma and leaving the cochlear implant in situ during acquisition in order to quantify the volume of fibrosis around the implant [ 46 , 47 ]. Although the resolution is not as good as with confocal microscopy, cellular evaluation is also possible with immunostaining of hair cells, neurons, and fibrosis [ 46 , 47 ].…”

“…It allows three-dimensional analysis of whole cochlear samples without dissection trauma and leaving the cochlear implant in situ during acquisition in order to quantify the volume of fibrosis around the implant [ 46 , 47 ]. Although the resolution is not as good as with confocal microscopy, cellular evaluation is also possible with immunostaining of hair cells, neurons, and fibrosis [ 46 , 47 ]. These imaging investigations (confocal microscopy and light sheet) require transparent samples.…”

“…These imaging investigations (confocal microscopy and light sheet) require transparent samples. With the development of these imaging techniques, numerous clearing protocols have been developed in recent years and adapted to cochlear clarification [ 46 , 47 , 48 ].…”