High spatial-resolution imaging of label-free<i>in vivo</i>protein aggregates by VISTA

Lin, Li-En; Miao, Kun; Qian, Chenxi; Lu, Wei

doi:10.1039/d1an00060h

Cited by 11 publications

(13 citation statements)

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“…Additionally, hyperspectral SRS , adds another layer of information for subcellular spectral analysis. With these capabilities, label-free SRS paves the way for various applications, including sensing environmental cues, studying lipid metabolism and identifying druggable targets, − tracking drug delivery and distribution, ,− multicolor cell sorting, , fast diagnosis of tumors, , and investigation of amyloid plaques in neurodegenerative diseases. , The current imaging speed, throughput, and detection sensitivity are still being continuously improved with rapid instrumental innovations. − In parallel, emerging data processing approaches, particularly machine learning algorithms, further upgrade image quality and enable data mining with rich chemical information. ,,− …”

Section: Label-free Vibrational Imagingmentioning

confidence: 99%

Bringing Vibrational Imaging to Chemical Biology with Molecular Probes

Wang

2022

ACS Chem. Biol.

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As an emerging optical imaging modality, stimulated Raman scattering (SRS) microscopy provides invaluable opportunities for chemical biology studies using its rich chemical information. Through rapid progress over the past decade, the development of Raman probes harnessing the chemical biology toolbox has proven to play a key role in advancing SRS microscopy and expanding biological applications. In this perspective, we first discuss the development of biorthogonal SRS imaging using small tagging of triple bonds or isotopes and highlight their unique advantages for metabolic pathway analysis and microbiology investigations. Potential opportunities for chemical biology studies integrating small tagging with SRS imaging are also proposed. We next summarize the current designs of highly sensitive and super-multiplexed SRS probes, as well as provide future directions and considerations for next-generation functional probe design. These rationally designed SRS probes are envisioned to bridge the gap between SRS microscopy and chemical biology research and should benefit their mutual development.

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Section: Label-free Vibrational Imagingmentioning

confidence: 99%

Bringing Vibrational Imaging to Chemical Biology with Molecular Probes

Wang

2022

ACS Chem. Biol.

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“…An interesting technique called VISTA combines label-free SRS imaging of protein with a special ExM protocol named magnified analysis of the proteome (MAP). [29,55] Compared to VISTA, MAGNIFIERS demonstrates expansion SRS microscopy on a much broader range of biological specimens. [30] Foremost, unlike VISTA that requires the acrylamide co-fixation, [26,29] MAGNIFIERS is not limited to soft tissue and features general sample compatibility across a broad range of tissue types and fixation methods.…”

Section: Discussionmentioning

confidence: 99%

Super‐Resolution Vibrational Imaging Using Expansion Stimulated Raman Scattering Microscopy

et al. 2022

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Stimulated Raman scattering (SRS) microscopy is an emerging technology that provides high chemical specificity for endogenous biomolecules and can circumvent common constraints of fluorescence microscopy including limited capabilities to probe small biomolecules and difficulty resolving many colors simultaneously. However, the resolution of SRS microscopy remains governed by the diffraction limit. To overcome this, a new technique called molecule anchorable gel‐enabled nanoscale Imaging of Fluorescence and stimulated Raman scattering microscopy (MAGNIFIERS) that integrates SRS microscopy with expansion microscopy (ExM) is described. MAGNIFIERS offers chemical‐specific nanoscale imaging with sub‐50 nm resolution and has scalable multiplexity when combined with multiplex Raman probes and fluorescent labels. MAGNIFIERS is used to visualize nanoscale features in a label‐free manner with CH vibration of proteins, lipids, and DNA in a broad range of biological specimens, from mouse brain, liver, and kidney to human lung organoid. In addition, MAGNIFIERS is applied to track nanoscale features of protein synthesis in protein aggregates using metabolic labeling of small metabolites. Finally, MAGNIFIERS is used to demonstrate 8‐color nanoscale imaging in an expanded mouse brain section. Overall, MAGNIFIERS is a valuable platform for super‐resolution label‐free chemical imaging, high‐resolution metabolic imaging, and highly multiplexed nanoscale imaging, thus bringing SRS to nanoscopy.

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“… 51 VISTA retains most endogenous proteins and removes excessive scattering lipids, optimizing conditions of cell and tissue samples for volumetric SRS imaging. Equipped with the machine-learning suite with the “ground truth” training data from immunolabeling fluorescence, VISTA provides label-free spatial predictions of the nucleus, blood vessels, neuronal cells, and dendrites in mouse brain tissues for multiplex imaging and microenvironment phenotyping with high accuracy 52 ( Figure 3 b).…”

Section: Coherent Raman Scattering: Srs and Cars Microscopymentioning

confidence: 99%

Toward the Next Frontiers of Vibrational Bioimaging

Wang

Lee

2023

Chemical & Biomedical Imaging

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Chemical imaging based on vibrational contrasts can extract molecular information entangled in complex biological systems. To this end, nonlinear Raman scattering microscopy, mid-infrared photothermal (MIP) microscopy, and atomic force microscopy (AFM)-based force-detected photothermal microscopies are emerging with better chemical sensitivity, molecular specificity, and spatial resolution than conventional vibrational methods. Their utilization in bioimaging applications has provided biological knowledge in unprecedented detail. This Perspective outlines key methodological developments, bioimaging applications, and recent technical innovations of the three techniques. Representative biological demonstrations are also highlighted to exemplify the unique advantages of obtaining vibrational contrasts. With years of effort, these three methods compose an expanding vibrational bioimaging toolbox to tackle specific bioimaging needs, benefiting many biological investigations with rich information in both label-free and labeling manners. Each technique will be discussed and compared in the outlook, leading to possible future directions to accommodate growing needs in vibrational bioimaging.

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High spatial-resolution imaging of label-freein vivoprotein aggregates by VISTA

Abstract: Amyloid aggregation, formed by aberrant proteins, is a pathological hallmark for neurodegenerative diseases, including Alzheimer’s disease and Huntington’s disease. High-resolution holistic mapping of the fine structures from these aggregates should...

Cited by 11 publications

References 50 publications

Bringing Vibrational Imaging to Chemical Biology with Molecular Probes

Bringing Vibrational Imaging to Chemical Biology with Molecular Probes

Super‐Resolution Vibrational Imaging Using Expansion Stimulated Raman Scattering Microscopy

Toward the Next Frontiers of Vibrational Bioimaging

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