Electron tomograms from chromatoid body in mouse haploid round spermatids

Data includes ET tilt series and reconstructions from testis tissue sample 2013.

Testis samples were fixed in 5% glutaraldehyde, post-fixed with osmium tetroxide, stained in UA-en-bloc, dehydrated in ethanol series and gradually embedded into epoxy. Serial semithick 250 nm sections were prepared and picked on single slot grids. Colloidal gold particles of 10 nm in diameter were placed below the sections to serve as fiducial markers for alignment of the tomograms. Dual axis tilt series were acquired using SerialEM software (http://bio3d.colorado.edu/SerialEM) running on a Tecnai FEG 20 microscope operating at 200 kV. Images from three consecutive sections were recorded at 1° intervals at 11.5k nominal magnification providing a 2x binned pixel size of 1.94 nm. IMOD software (http://bio3d.colorado.edu/imod) was used to create 3D reconstructions from the tilt series. The images were segmented using Microscopy Image Browser-software (https://mib.helsinki.fi) and 3D-rendering of the segmented structures was performed with BioimageXD version 1 (Kankaanpää et al., 2012)

Retromer vesicles interact with RNA granules in haploid male germ cells. Da Ros M, Hirvonen N, Olotu O, Toppari J, Kotaja N. Mol Cell Endocrinol. 2015 Feb 5;401:73‐83. doi: 10.1016/j.mce.2014.11.026. Epub 2014 Dec 5. PMID: 25486514
Fig. 2 and Supplementary Movie S2

Da Ros M, Lehtiniemi T, Olotu O, Fischer D, Zhang FP, Vihinen H, Jokitalo E, Sironen A, Toppari J, Kotaja N. FYCO1 and autophagy control the integrity of the haploid male germ cell-specific RNP granules. Autophagy. 2017 Feb;13(2):302-321. doi: 10.1080/15548627.2016.1261319. Epub 2016 Dec 8. PMID: 27929729; PMCID: PMC5324852.