Journal article

A new method for three-dimensional immunofluorescence study of the cochlea

Kate M Brody, Amy J Hampson, Hyun-jung Cho, Prudence Johnson, Stephen J O'Leary

HEARING RESEARCH | ELSEVIER | Published : 2020

Abstract

Visualisation of cochlear histopathology in three-dimensions has been long desired in the field of hearing research. This paper outlines a technique that has made this possible and shows a research application in the field of hearing protection after cochlear implantation. The technique utilises robust immunofluorescent labelling followed by effective tissue clearing and fast image acquisition using Light Sheet Microscopy. We can access the health of individual components by immunofluorescent detection of proteins such as myosin VIIa to look at cochlear hair cells, NaKATPase alpha 3 to look at spiral ganglion neurons, and IBA1 to look at macrophages within a single cochlea, whilst maintainin..

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Grants

Awarded by National Health and Medical Research Council (Australia)


Funding Acknowledgements

The authors would like to thank Samantha Moreland for animal husbandry assistance, Helen Feng for electrode manufacture, Dr Gabriela Segal from the Biological Optical Microscopy Platform, University of Melbourne for imaging assistance, and Dr Aaron Collins for assistance in preparing figures. This work was funded by the National Health and Medical Research Council (Australia; GNT1125970), the Garnett Passe and Rodney Williams Memorial Foundation, and a generous bequest from the family of the late Gordon Darling, AC.