Spatial Proteomics: A Gateway to Understanding Cell Biology

Borner, Georg H. H.

doi:10.1002/pmic.201900328

Cited by 3 publications

(2 citation statements)

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“…Among the emerging spatial technologies, spatial proteomics provides the direct functional readouts of cellular/tissue phenotypes and thus has undergone rapid development in the past decade. ,,− Current spatial proteomics can be classified into two different approaches depending on whether an antibody is used or not. Antibody-based multiplex imaging approaches, such as CODEX (Co-detection by indexing), IBEX (Iterative bleaching extends multiplexity), IMC (Imaging Mass Cytometry), , and MIBI (Multiplexed Ion Beam Imaging), have enabled mapping of >40 proteins on the same tissue samples at subcellular resolution.…”

Section: Introductionmentioning

confidence: 99%

Spatial Proteomics toward Subcellular Resolution by Coupling Deep Ultraviolet Laser Ablation with Nanodroplet Sample Preparation

Xiang,

Liyu,

Kwon

et al. 2023

ACS Meas. Sci. Au

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Multiplexed molecular profiling of tissue microenvironments, or spatial omics, can provide critical insights into cellular functions and disease pathology. The coupling of laser microdissection with mass spectrometry-based proteomics has enabled deep and unbiased mapping of >1000 proteins. However, the throughput of laser microdissection is often limited due to tedious two-step procedures, sequential laser cutting, and sample collection. The two-step procedure also hinders the further improvement of spatial resolution to <10 μm as needed for subcellular proteomics. Herein, we developed a high-throughput and high-resolution spatial proteomics platform by seamlessly coupling deep ultraviolet (DUV) laser ablation (LA) with nanoPOTS (Nanodroplet Processing in One pot for Trace Samples)-based sample preparation. We demonstrated the DUV-LA system can quickly isolate and collect tissue samples at a throughput of ∼30 spots/min and a spatial resolution down to 2 μm from a 10 μm thick human pancreas tissue section. To improve sample recovery, we developed a proximity aerosol collection approach by placing DMSO droplets close to LA spots. We demonstrated the DUV-LA-nanoPOTS platform can detect an average of 1312, 1533, and 1966 proteins from ablation spots with diameters of 7, 13, and 19 μm, respectively. In a proof-of-concept study, we isolated and profiled two distinct subcellular regions of the pancreas tissue revealed by hematoxylin and eosin (H&E) staining. Quantitative proteomics revealed proteins specifically enriched to subcellular compartments.

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Section: Introductionmentioning

confidence: 99%

Spatial Proteomics toward Subcellular Resolution by Coupling Deep Ultraviolet Laser Ablation with Nanodroplet Sample Preparation

Xiang,

Liyu,

Kwon

et al. 2023

ACS Meas. Sci. Au

View full text Add to dashboard Cite

show abstract

“…Research on the localization and quantification of proteins on the subcellular scale belongs to the discipline known as spatial proteomics, a branch of proteomics (7). Now, two methods are available for spatial proteomics analysis: mass spectrometry (MS)-based organelle profiling and imaging-based protein localization approaches (8)(9)(10)(11). MS can identify and quantify proteins with high throughput, making it a powerful tool for proteomics research (12)(13)(14).…”

Section: Introductionmentioning

confidence: 99%