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Bimodal endocytic probe for three-dimensional correlative light and electron microscopy

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Publication date

2022-05-23

Authors

Fermie, Job
de Jager, Leanne
Foster, Helen E
Veenendaal, TinekeISNI 0000000394529994
de Heus, Cecilia
van Dijk, Suzanne
ten Brink, Corlinda
Oorschot, Viola
Yang, Lin
Li, Wei

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DOI

https://doi.org/10.1016/j.crmeth.2022.100220

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Article

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Abstract

We present a bimodal endocytic tracer, fluorescent BSA-gold (fBSA-Au), as a fiducial marker for 2D and 3D correlative light and electron microscopy (CLEM) applications. fBSA-Au consists of colloidal gold (Au) particles stabilized with fluorescent BSA. The conjugate is efficiently endocytosed and distributed throughout the 3D endolysosomal network of cells and has an excellent visibility in both fluorescence microscopy (FM) and electron microscopy (EM). We demonstrate that fBSA-Au facilitates rapid registration in several 2D and 3D CLEM applications using Tokuyasu cryosections, resin-embedded material, and cryoelectron microscopy (cryo-EM). Endocytosed fBSA-Au benefits from a homogeneous 3D distribution throughout the endosomal system within the cell, does not obscure any cellular ultrastructure, and enables accurate (50-150 nm) correlation of fluorescence to EM data. The broad applicability and visibility in both modalities makes fBSA-Au an excellent endocytic fiducial marker for 2D and 3D (cryo)CLEM applications.

Keywords

correlative light and electron microscopy, cryoelectron microscopy, electron tomography, endolysosomal system, nanogold fiducials, volume-electron microscopy, Genetics, Biochemistry, Genetics and Molecular Biology (miscellaneous), Biochemistry, Radiology Nuclear Medicine and imaging, Biotechnology, Computer Science Applications, Journal Article

Citation

Fermie, J, de Jager, L, Foster, H E, Veenendaal, T, de Heus, C, van Dijk, S, Ten Brink, C, Oorschot, V, Yang, L, Li, W, Müller, W H, Howes, S, Carter, A P, Förster, F, Posthuma, G, Gerritsen, H C, Klumperman, J & Liv, N 2022, 'Bimodal endocytic probe for three-dimensional correlative light and electron microscopy', Cell reports methods, vol. 2, no. 5, 100220, pp. 1-22. https://doi.org/10.1016/j.crmeth.2022.100220

URI

https://dspace.library.uu.nl/handle/1874/446518