It's a Small, Small World

Guérin, Christopher J.; Liv, Nalan; Klumperman, Judith

doi:10.1002/9781119086420.ch1

Cited by 3 publications

(2 citation statements)

References 111 publications

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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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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.

View full text Add to dashboard Cite

show abstract

“…Although single-cell and ‘omic technologies are relatively young fields that have been developing the past 20 years [ 1 ], single-cell imaging in the form of microscopy can be traced back to the 1600s, when Robert Hooke first described the small rectangular compartments he observed with his microscope in cork bark as ‘cells' [ 2 ]. Little did he know that what reminded him of small rooms in which monks slept in, were in fact the smallest, single units of life, and that his early work was part of the birth of modern microscopy and to an extension, single-cell imaging.…”

Section: Introductionmentioning

confidence: 99%

From imaging a single cell to implementing precision medicine: an exciting new era

Karacosta

2021

Emerging Topics in Life Sciences

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In the age of high-throughput, single-cell biology, single-cell imaging has evolved not only in terms of technological advancements but also in its translational applications. The synchronous advancements of imaging and computational biology have produced opportunities of merging the two, providing the scientific community with tools towards observing, understanding, and predicting cellular and tissue phenotypes and behaviors. Furthermore, multiplexed single-cell imaging and machine learning algorithms now enable patient stratification and predictive diagnostics of clinical specimens. Here, we provide an overall summary of the advances in single-cell imaging, with a focus on high-throughput microscopy phenomics and multiplexed proteomic spatial imaging platforms. We also review various computational tools that have been developed in recent years for image processing and downstream applications used in biomedical sciences. Finally, we discuss how harnessing systems biology approaches and data integration across disciplines can further strengthen the exciting applications and future implementation of single-cell imaging on precision medicine.

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Sample preparation strategies for efficient correlation of 3D SIM and soft X-ray tomography data at cryogenic temperatures

et al. 2021

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This protocol describes sample preparation strategies for correlative 3D cryo-SIM and cryo soft X-ray tomography. In addition, the authors provide a direct comparison and recommendations regarding the selection and use of fiducials for 3D correlation. TWEET A new protocol for sample preparation and fiducial selection for correlative cryo-SIM and cryo-soft X-ray tomography. #CLXT #cryoimaging #cellstructure #correlativeimaging COVER TEASER Correlative cryo-SIM and cryo-soft X-ray tomographyAbstract 3D correlative microscopy methods have revolutionised biomedical research allowing the acquisition of multi-dimensional information to gain an in-depth understanding of biological systems. With the advent of relevant cryo-preservation methods, correlative imaging of cryogenically preserved samples has led to nanometre resolution imaging (2-50 nm) under harsh imaging regimes such as electron and soft X-ray tomography. These methods have now been combined with conventional and super resolution fluorescence imaging at cryogenic temperatures to augment information content from a given sample resulting in the immediate requirement for protocols that facilitate hassle-free unambiguous cross correlation between microscopes. We present here sample preparation strategies and a direct comparison of different working fiducialisation regimes that facilitate 3D correlation of cryo-structured illumination microscopy and cryo-soft X-ray tomography. Our protocol has been tested at two synchrotron beamlines (B24 at Diamond Light Source in the UK and BL09 Mistral at ALBA in Spain) and has led to the development of a decision aid that facilitates experimental design with the strategic use of markers based on project requirements. This protocol takes between 1.5 hours and 3.5 days to complete, depending on the cell populations used (adherent cells may require several days to grow on sample carriers).

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It's a Small, Small World

Cited by 3 publications

References 111 publications

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

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

From imaging a single cell to implementing precision medicine: an exciting new era

Sample preparation strategies for efficient correlation of 3D SIM and soft X-ray tomography data at cryogenic temperatures

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