Relocation is the key to successful correlative fluorescence and scanning electron microscopy

Cheng, Delfine; Shami, Gerald J.; Morsch, Marco; Huynh, Minh; Trimby, Patrick; Braet, Filip

doi:10.1016/bs.mcb.2017.03.013

Cited by 6 publications

(3 citation statements)

References 52 publications

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“…Instead, only a few macrophage-like cells with a distinct morphology to Kupffer cells were scattered deep within the liver parenchymal cords. 3,4 That said, an identical structural and functional relationship was found between the liver sinusoidal endothelial lining, Space of Disse and hepatocytes. 5 Undoubtedly, the use of zebrafish is a powerful experimental alternative that has already revolutionised biomedical research by providing new disease models to study cancer, drug metabolism, infection and vascular-related changes in various organ systems.…”

Section: E T T E R T O T H E E D I T O R Zebrafish: An Emerging and E...mentioning

confidence: 87%

Zebrafish: An emerging and expedient model for liver studies?

Shami

Cheng

Morsch

et al. 2023

Liver International

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Section: E T T E R T O T H E E D I T O R Zebrafish: An Emerging and E...mentioning

confidence: 87%

Zebrafish: An emerging and expedient model for liver studies?

Shami

Cheng

Morsch

et al. 2023

Liver International

Self Cite

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“…Correlation of light and EM as a method became substantially more elaborate with the introduction of serial block-face sectioning. Now, a 3D-LM data set with precise reference coordinates serves a basis (Lucas et al, 2012; Karreman et al, 2016; Cheng et al, 2017; Lees et al, 2017; Russell et al, 2017). For standard EM preparations, specimens have to be cut into small pieces, which is accompanied by a significant loss of the 3D-context.…”

Section: Discussionmentioning

confidence: 99%

From Light Microscopy to Analytical Scanning Electron Microscopy (SEM) and Focused Ion Beam (FIB)/SEM in Biology: Fixed Coordinates, Flat Embedding, Absolute References

Luckner

Wanner

2018

Microsc Microanal

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Correlative light and electron microscopy (CLEM) has been in use for several years, however it has remained a costly method with difficult sample preparation. Here, we report a series of technical improvements developed for precise and cost-effective correlative light and scanning electron microscopy (SEM) and focused ion beam (FIB)/SEM microscopy of single cells, as well as large tissue sections. Customized coordinate systems for both slides and coverslips were established for thin and ultra-thin embedding of a wide range of biological specimens. Immobilization of biological samples was examined with a variety of adhesives. For histological sections, a filter system for flat embedding was developed. We validated ultra-thin embedding on laser marked slides for efficient, high-resolution CLEM. Target cells can be re-located within minutes in SEM without protracted searching and correlative investigations were reduced to a minimum of preparation steps, while still reaching highest resolution. The FIB/SEM milling procedure is facilitated and significantly accelerated as: (i) milling a ramp becomes needless, (ii) significant re-deposition of milled material does not occur; and (iii) charging effects are markedly reduced. By optimizing all technical parameters FIB/SEM stacks with 2 nm iso-voxels were achieved over thousands of sections, in a wide range of biological samples.

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“…It has long been recognized that combining several modalities to observe a given sample can be very useful in science. In biology, the initial focus has been in combining electron and fluorescence microscopy [1][2][3]. The initial interest has expanded to the correlation of several other modalities [4,5], including atomic force microscopy (AFM) [6][7][8], super-resolution microscopy [7,9], and Raman microscopy [10].…”

Section: Introductionmentioning

confidence: 99%

An efficient solution for correlative microscopy and co-localized observations based on multiscale multimodal machine-readable nanoGPS tags

Acher

Nguyen

Podzorov

et al. 2021

Meas. Sci. Technol.

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For a long time, investigating the same regions of interest of a sample with different instruments has been recognized as a very useful approach in various scientific fields. This paper presents an original solution for spotting the same points of interest with a high degree of accuracy and simplicity using different microscopes. It is based on small patterned tags fixed to the samples or their substrates. The patterns include an image-based position-sensing technology, for which an image of a small part of the tag can be automatically converted to absolute coordinates and angular orientation. Taking a single snapshot of the tag with an imaging instrument provides a correspondence between the sample and the coordinates of the moving stage. Co-localized observations performed with scanning electron microscopes, optical microscopes, and Raman microscopes are presented. The accuracy is in the range of a few µm up to 20 µm, which is generally sufficient to remove any ambiguity between the observed objects. The different contributions to colocalization errors are investigated experimentally and it is shown that errors related to the tags are negligible and that the main source of error is related to the accuracy of the moving stages integrated into the microscopes. A straightforward estimation of the relocalization error can be performed. It is believed that this solution will save researchers time and facilitate cooperation between laboratories.

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Relocation is the key to successful correlative fluorescence and scanning electron microscopy

Cited by 6 publications

References 52 publications

Zebrafish: An emerging and expedient model for liver studies?

Zebrafish: An emerging and expedient model for liver studies?

From Light Microscopy to Analytical Scanning Electron Microscopy (SEM) and Focused Ion Beam (FIB)/SEM in Biology: Fixed Coordinates, Flat Embedding, Absolute References

An efficient solution for correlative microscopy and co-localized observations based on multiscale multimodal machine-readable nanoGPS tags

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