Cochleae from wild-type (Triobp+/+), heterozygous (TriobpΔEx9-10/+), and homozygous (TriobpΔEx9-10/ΔEx9-10) mice at P6 and P7 were used. FIB-SEM stacks were imaged using a FEI Helios 660 Nanolab system in “Slice and View” mode with a backscattered electron detector. For detailed descriptions of FIB-SEM sample preparation, data collection, and analysis of FIB-SEM stacks, see SI Appendix, Supplementary Materials and Methods .
Helios 660 nanolab system
Manufactured by Thermo Fisher Scientific
Sourced in Germany
The Helios 660 Nanolab system is a focused ion beam (FIB) scanning electron microscope (SEM) designed for high-resolution imaging and analysis of nanostructures. It combines a high-resolution field emission SEM column with a precise ion beam column for milling, etching, and site-specific sample preparation. The system provides nanometer-scale 3D characterization and manipulation capabilities for a wide range of materials and applications.
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Lab products found in correlation
2 protocols using helios 660 nanolab system
1
FIB-SEM Imaging of Triobp Mutant Cochleae
2
Cryo-Preparation for Organ of Corti Imaging
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Organ of Corti explants were fixed overnight in 3% glutaraldehyde in 0.15 M sodium cacodylate buffer (pH 7.4) at 4°C, rinsed thoroughly with sodium cacodylate buffer, and post-fixed for 1 hr in 1% tannic acid. Then, the samples were rinsed with distilled water and cryoprotected by overnight incubations in 5, 10 and 30% glycerol solutions. The explants were then placed on 3 mm copper grids and plunge frozen in liquid Freon before being transferred to 1% uranyl acetate in methanol at −90°C in a Leica EM AFS2 freeze substitution machine. Methanol was exchanged for Lowicryl HM-20 resin and polymerized by long-wave UV radiation. All reagents were obtained from Electron Microscopy Sciences. The resin blocks were trimmed on an ultramicrotome (UC6, Leica, Wetzlar, Germany) and then milled with a focused ion beam and imaged in ‘Slice and View’ mode with a backscattered electron detector using the FEI Helios 660 Nanolab system.
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