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

Acher, O.; Nguyen, Thanhliem; Podzorov, Alexander; Leroy, Matthieu; Carles, Pierre-Alix; Legendre, Sébastien

doi:10.1088/1361-6501/abce39

Cited by 8 publications

(11 citation statements)

References 33 publications

(54 reference statements)

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“…Currently, our repositioning system can hardly achieve the accuracy of a nanometer. A similar report published recently [ 27 ] pointed out that stage accuracy, or instrument stability, could be the limiting factor for relocalization accuracy. In our experiment, we also found that the error delivered from the instrument stability was one of the main errors in the repositioning system, which was supported by the stage reproducibility and accuracy test executed in SEM and AFM independently (see in Table S3 ).…”

Section: Resultsmentioning

confidence: 86%

A Universal Positioning System for Coupling Characterization of SEM and AFM

et al. 2021

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Hyphenated techniques, providing comprehensive information in various aspects such as constituent, structure, functional group, and morphology, play an important role in scientific research. Nowadays, coupling characterization of the same position in microscale is in great need in the field of nanomaterial research and exploration. In this article, a new hyphenated technique was developed to facilitate the coupling characterization of atomic force microscope (AFM) and scanning electron microscope (SEM) by designing a universal positioning system. The system consisted of a specimen holder with coordinate grids and a software for converting the coordinate values of the same point to fit SEM, specimen holder, and AFM system. In working condition, the coordinates of the labeled points and target position were firstly extracted from the SEM operation software, then converted into the numerical values adapted to the specimen holder itself, and finally transformed into the coordinates matching the AFM system. The experimental result showed that a retrieving rate of 96% was achieved for a spherical target with a diameter of 1 μm in a 30 μ m × 30 μ m square. The hyphenated technique is a universal, accurate, efficient, and financially feasible method in microanalysis field and has great application potential.

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Section: Resultsmentioning

confidence: 86%

A Universal Positioning System for Coupling Characterization of SEM and AFM

et al. 2021

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“…In addition to sample preparation, a special sample holder was developed. In order to find the ROIs in different devices, a nanoGPS Oxyo ® tag developed by Acher et al was used [33]. The technology allowed accurate and precise retrieval of the selected point on the sample, with precise calibration of the stage.…”

Section: Discussionmentioning

confidence: 99%

Characterizing Intraindividual Podocyte Morphology In Vitro with Different Innovative Microscopic and Spectroscopic Techniques

Kraus¹,

Rose

Krüger

et al. 2023

Cells

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Podocytes are critical components of the glomerular filtration barrier, sitting on the outside of the glomerular basement membrane. Primary and secondary foot processes are characteristic for podocytes, but cell processes that develop in culture were not studied much in the past. Moreover, protocols for diverse visualization methods mostly can only be used for one technique, due to differences in fixation, drying and handling. However, we detected by single-cell RNA sequencing (scRNAseq) analysis that cells reveal high variability in genes involved in cell type-specific morphology, even within one cell culture dish, highlighting the need for a compatible protocol that allows measuring the same cell with different methods. Here, we developed a new serial and correlative approach by using a combination of a wide variety of microscopic and spectroscopic techniques in the same cell for a better understanding of podocyte morphology. In detail, the protocol allowed for the sequential analysis of identical cells with light microscopy (LM), Raman spectroscopy, scanning electron microscopy (SEM) and atomic force microscopy (AFM). Skipping the fixation and drying process, the protocol was also compatible with scanning ion-conductance microscopy (SICM), allowing the determination of podocyte surface topography of nanometer-range in living cells. With the help of nanoGPS Oxyo®, tracking concordant regions of interest of untreated podocytes and podocytes stressed with TGF-β were analyzed with LM, SEM, Raman spectroscopy, AFM and SICM, and revealed significant morphological alterations, including retraction of podocyte process, changes in cell surface morphology and loss of cell-cell contacts, as well as variations in lipid and protein content in TGF-β treated cells. The combination of these consecutive techniques on the same cells provides a comprehensive understanding of podocyte morphology. Additionally, the results can also be used to train automated intelligence networks to predict various outcomes related to podocyte injury in the future.

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“…Another family of position sensing technique is based on imaging a scale with engineered patterns and retrieving the position and orientation from the image [17][18][19][20][21][22] . It has been used to perform in-plane position determination in various contexts, including robotics 17 , machine vision 23 , and microscopy 13,24 . Our group has developed a particular family of position-encoding patterns and reading software, termed nanoGPS.…”

Section: Introductionmentioning

confidence: 99%

Identifying and fixing in-plane positioning and stability issues on a microscope using nanoGPS OxyO scales

Acher,

de Abreu,

Grigoriev

et al. 2023

Preprint

Self Cite

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Investigations of the in-plane positioning capabilities of microscopes using commercially available nanoGPS OxyO scales are presented. The scales have patterns that contain absolute position information, and nanoGPS software accurately determines the in-plane position from the scale images captured by the microscope camera. This makes in-plane positioning experiments simple and fast. We investigated different microscopy systems and found that positioning performance is a system issue that is not determined solely by the stage performance. In some cases, our experiments revealed software or hardware glitches that limited the positioning performance, which we easily fixed. We have also shown that it is possible to investigate vibrations using this approach and quantify their impact on image blurring. This is, for example, useful for experimentally determining the settling time after a stage movement.

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

Cited by 8 publications

References 33 publications

A Universal Positioning System for Coupling Characterization of SEM and AFM

A Universal Positioning System for Coupling Characterization of SEM and AFM

Characterizing Intraindividual Podocyte Morphology In Vitro with Different Innovative Microscopic and Spectroscopic Techniques

Identifying and fixing in-plane positioning and stability issues on a microscope using nanoGPS OxyO scales

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