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

Job Fermie; Leanne de Jager; Helen E Foster; Tineke Veenendaal; Cecilia de Heus; Suzanne van Dijk; Corlinda Ten Brink; Viola Oorschot; Lin Yang; Wei Li; Wally H Müller; Stuart Howes; Andrew P Carter; Friedrich Förster; George Posthuma; Hans C Gerritsen; Judith Klumperman; Nalan Liv

doi:10.1016/j.crmeth.2022.100220

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

Job Fermie, Leanne de Jager, Helen E Foster, Tineke Veenendaal, Cecilia de Heus, Suzanne van Dijk, Corlinda Ten Brink, Viola Oorschot, Lin Yang, Wei Li, Wally H Müller, Stuart Howes, Andrew P Carter, Friedrich Förster, George Posthuma, Hans C Gerritsen, Judith Klumperman, Nalan Liv

Research output: Contribution to journal › Article › Academic › peer-review

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.

Original language	English
Article number	100220
Number of pages	23
Journal	Cell Reports Methods
Volume	2
Issue number	5
DOIs	https://doi.org/10.1016/j.crmeth.2022.100220
Publication status	Published - 23 May 2022

Bibliographical note

Funding Information:
We acknowledge funding from the Netherlands Organisation for Scientific Research (NWO), Stichting voor de Technische Wetenschappen (Technology Foundation STW), grant 12715 (to H.G. and J.K.) and the Netherlands Organisation for Health Research and Development (ZonMW) grant 40-00812-98-16006 (to J.K.). The microscopy infrastructure in this work is partially subsidized by the Roadmap for Large-Scale Research Infrastructure (NEMI) of Netherlands Organisation for Scientific Research (grant number 184.034.014 ) (to J.K.). The Cell Microscopy Core of UMC Utrecht is part of the Dutch Correlative Light Electron Microscopy node of EuroBioImaging.

Publisher Copyright:
© 2022 The Author(s)

Keywords

correlative light and electron microscopy
cryoelectron microscopy
electron tomography
endolysosomal system
nanogold fiducials
volume-electron microscopy

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1-s2.0-S2667237522000832-main (1)Final published version, 8.45 MBLicence: CC BY

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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, 2(5), Article 100220. https://doi.org/10.1016/j.crmeth.2022.100220

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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",

author = "Job Fermie and {de Jager}, Leanne and Foster, {Helen E} and Tineke Veenendaal and {de Heus}, Cecilia and {van Dijk}, Suzanne and {Ten Brink}, Corlinda and Viola Oorschot and Lin Yang and Wei Li and M{\"u}ller, {Wally H} and Stuart Howes and Carter, {Andrew P} and Friedrich F{\"o}rster and George Posthuma and Gerritsen, {Hans C} and Judith Klumperman and Nalan Liv",

note = "Funding Information: We acknowledge funding from the Netherlands Organisation for Scientific Research (NWO), Stichting voor de Technische Wetenschappen (Technology Foundation STW), grant 12715 (to H.G. and J.K.) and the Netherlands Organisation for Health Research and Development (ZonMW) grant 40-00812-98-16006 (to J.K.). The microscopy infrastructure in this work is partially subsidized by the Roadmap for Large-Scale Research Infrastructure (NEMI) of Netherlands Organisation for Scientific Research (grant number 184.034.014 ) (to J.K.). The Cell Microscopy Core of UMC Utrecht is part of the Dutch Correlative Light Electron Microscopy node of EuroBioImaging. Publisher Copyright: {\textcopyright} 2022 The Author(s)",

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

TY - JOUR

T1 - Bimodal endocytic probe for three-dimensional correlative light and electron microscopy

AU - Fermie, Job

AU - de Jager, Leanne

AU - Foster, Helen E

AU - Veenendaal, Tineke

AU - de Heus, Cecilia

AU - van Dijk, Suzanne

AU - Ten Brink, Corlinda

AU - Oorschot, Viola

AU - Yang, Lin

AU - Li, Wei

AU - Müller, Wally H

AU - Howes, Stuart

AU - Carter, Andrew P

AU - Förster, Friedrich

AU - Posthuma, George

AU - Gerritsen, Hans C

AU - Klumperman, Judith

AU - Liv, Nalan

N1 - Funding Information: We acknowledge funding from the Netherlands Organisation for Scientific Research (NWO), Stichting voor de Technische Wetenschappen (Technology Foundation STW), grant 12715 (to H.G. and J.K.) and the Netherlands Organisation for Health Research and Development (ZonMW) grant 40-00812-98-16006 (to J.K.). The microscopy infrastructure in this work is partially subsidized by the Roadmap for Large-Scale Research Infrastructure (NEMI) of Netherlands Organisation for Scientific Research (grant number 184.034.014 ) (to J.K.). The Cell Microscopy Core of UMC Utrecht is part of the Dutch Correlative Light Electron Microscopy node of EuroBioImaging. Publisher Copyright: © 2022 The Author(s)

PY - 2022/5/23

Y1 - 2022/5/23

N2 - 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.

AB - 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.

KW - correlative light and electron microscopy

KW - cryoelectron microscopy

KW - electron tomography

KW - endolysosomal system

KW - nanogold fiducials

KW - volume-electron microscopy

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U2 - 10.1016/j.crmeth.2022.100220

DO - 10.1016/j.crmeth.2022.100220

M3 - Article

C2 - 35637912

SN - 2667-2375

VL - 2

JO - Cell Reports Methods

JF - Cell Reports Methods

IS - 5

M1 - 100220

ER -

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

Abstract

Bibliographical note

Keywords

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