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Investigation of intact mouse cochleae using two‐photon laser scanning microscopy
Author(s) -
Bae Seong Hoon,
Kwak Sang Hyun,
Choe Young Ho,
Hyun YoungMin,
Choi Jae Young,
Jung Jinsei
Publication year - 2020
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.23515
Subject(s) - spiral ligament , cochlea , immunolabeling , clearance , staining , anatomy , confocal laser scanning microscopy , confocal , biomedical engineering , fluorescence , fluorescence microscope , stereocilia (inner ear) , chemistry , biophysics , pathology , hair cell , biology , organ of corti , immunohistochemistry , medicine , optics , physics , urology
Objectives The investigation of cochlear hair cells and lateral wall is a time‐consuming and labor‐intensive process. However, it is a mandatory experiment in audiology research. Here we suggest a novel method for investigating the inner ear microstructures from intact cochleae using two‐photon laser scanning microscopy (TPLSM). This technique guarantees fewer artifacts and technical simplicity. Methods Using TPLSM, we investigated the whole mount cochleae, decalcified cochleae, and cleared cochleae of wild type C57BL/6 mice. CX3CR1 +/GFP mice were used to investigate the feasibility of visualizing cellular structures in the cochlear spiral ligament. All samples were investigated without staining. Results Endogenous fluorescence emission from the outer hair cells was strong enough to be distinguished from the other structures in all samples. From the single apical view, 50 and 90% of the whole hair cells of the decalcified cochleae and cleared cochleae, respectively, could be visualized without staining using TPLSM. Capillary structure of stria vascularis and spiral ligament could be visualized by endogenous fluorescence without staining. Conclusion We successfully investigated the hair cells and lateral wall of mouse cochleae using TPLSM without using staining or any destructive procedures. This method is easier, faster, and more reliable than conventional methods.