High-precision correlative fluorescence and electron cryo microscopy using two independent alignment markers
Abstract
Correlative light and electron microscopy (CLEM) is an emerging technique which combines functional information provided by fluorescence microscopy (FM) with the high-resolution structural information of electron microscopy (EM). So far, correlative cryo microscopy of frozen-hydrated samples has not reached better than micrometre range accuracy. Here, a method is presented that enables the correlation between fluorescently tagged proteins and electron cryo tomography (cryoET) data with nanometre range precision. Specifically, thin areas of vitrified whole cells are examined by correlative fluorescence cryo microscopy (cryoFM) and cryoET. Novel aspects of the presented cryoCLEM workflow not only include the implementation of two independent electron dense fluorescent markers to improve the precision of the alignment, but also the ability of obtaining an estimate of the correlation accuracy for each individual object of interest. The correlative workflow from plunge-freezing to cryoET is detailed step-by-step for the example of locating fluorescence-labelled adenovirus particles trafficking inside a cell.
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BibTeX
@article{schellenberger2026highprecision,
title = {{High-precision correlative fluorescence and electron cryo microscopy using two independent alignment markers}},
author = {Pascale Schellenberger and Rainer Kaufmann and C. Alistair Siebert and Christoph Hagen and Harald Wodrich and Kay Grünewald},
journal = {Ultramicroscopy},
year = {2026},
month = oct,
doi = {10.1016/j.ultramic.2013.10.011},
url = {https://hal.science/hal-05784099v1},
note = {CC BY},
}
RIS
TY - JOUR
TI - High-precision correlative fluorescence and electron cryo microscopy using two independent alignment markers
AU - Pascale Schellenberger
AU - Rainer Kaufmann
AU - C. Alistair Siebert
AU - Christoph Hagen
AU - Harald Wodrich
AU - Kay Grünewald
PY - 2026
DA - 2026/10/07
JO - Ultramicroscopy
DO - 10.1016/j.ultramic.2013.10.011
UR - https://hal.science/hal-05784099v1
AB - Correlative light and electron microscopy (CLEM) is an emerging technique which combines functional information provided by fluorescence microscopy (FM) with the high-resolution structural information of electron microscopy (EM). So far, correlative cryo microscopy of frozen-hydrated samples has not reached better than micrometre range accuracy. Here, a method is presented that enables the correlation between fluorescently tagged proteins and electron cryo tomography (cryoET) data with nanometre range precision. Specifically, thin areas of vitrified whole cells are examined by correlative fluorescence cryo microscopy (cryoFM) and cryoET. Novel aspects of the presented cryoCLEM workflow not only include the implementation of two independent electron dense fluorescent markers to improve the precision of the alignment, but also the ability of obtaining an estimate of the correlation accuracy for each individual object of interest. The correlative workflow from plunge-freezing to cryoET is detailed step-by-step for the example of locating fluorescence-labelled adenovirus particles trafficking inside a cell.
SN - 0304-3991
ER -