Source 1primary paper2025Chemical Science
Toolkit/SiNx solid-state nanopore single-cell protein analysis approach
SiNx solid-state nanopore single-cell protein analysis approach
Also known as: SiNx solid-state nanopores for single-molecule protein analysis from single cells
Taxonomy: Mechanism Branch / Component. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
The SiNx solid-state nanopore single-cell protein analysis approach combines glass nanopipette-based single-cell extraction with silicon nitride nanopore sensing for single-molecule protein analysis from complex cellular samples. In the cited Chemical Science study, this workflow identified the LOV2 protein and monitored its conformational changes directly from single-cell extracts.
Usefulness & Problems
Why this is useful
This approach is useful for probing proteins at the single-molecule level directly from individual cells rather than relying only on purified preparations. The reported ability to identify LOV2 and distinguish conformational behavior in single-cell extracts indicates utility for studying protein state in complex native-like cellular material.
Problem solved
It addresses the problem of measuring and identifying proteins from single cells using a solid-state nanopore platform despite the complexity of cellular samples. It also addresses the gap between purified-sample measurements and direct single-cell measurements, because the study reported significant differences between proteins measured from single cells and proteins obtained from purified samples.
Published Workflows
Exploring a solid-state nanopore approach for single-molecule protein detection from single cells
2025Objective: Develop and test a solid-state nanopore approach for direct single-molecule protein detection and conformational monitoring from single-cell extracts.
Why it works: The abstract states that a nanopore electrophoretic driver was fused to LOV2 to enhance capture efficiency, after which single-cell contents were extracted and analyzed with SiNx nanopores to identify the protein and monitor conformational changes.
electrophoretic-driver-assisted protein capturenanopore-based single-molecule protein sensingconformational change monitoringfusion construct designglass nanopipette-based single-cell extractionsolid-state nanopore measurement
Stages
- 1.Capture-enhancing fusion design(library_design)
The abstract states that the driver fusion was designed to enhance capture efficiency before nanopore analysis.
Selection: Design a nanopore electrophoretic driver protein fused to LOV2 to enhance target-protein capture efficiency.
- 2.Single-cell content extraction(functional_characterization)
This stage provides ex situ single-cell extracts for downstream nanopore protein analysis.
Selection: Directly extract contents of individual cells using glass nanopipette-based single-cell extraction.
- 3.Nanopore readout of single-cell extracts(confirmatory_validation)
This stage tests whether the overall approach can read out a model protein directly from single-cell extracts.
Selection: Use SiNx nanopores to identify LOV2 and monitor its conformational changes from single-cell extracts.
Steps
- 1.Design nanopore electrophoretic driver and fuse it to LOV2engineered fusion construct
Enhance capture efficiency of the target protein for nanopore analysis.
The abstract indicates this engineering step precedes single-cell analysis because improved capture is needed before nanopore measurement.
- 2.Extract contents of individual cells using glass nanopipette-based single-cell extractionsingle-cell sampling method
Obtain ex situ single-cell extracts for downstream nanopore protein analysis.
This step is performed after construct design to provide the single-cell material that will be analyzed by nanopores.
- 3.Use SiNx nanopores to identify LOV2 and monitor conformational changes from single-cell extractsnanopore analysis platform
Demonstrate direct single-molecule protein detection and conformational monitoring from single-cell extracts.
This is the downstream readout step applied after single-cell extraction to test the full approach on cellular material.
Taxonomy & Function
Primary hierarchy
Mechanism Branch
Component: A low-level protein part used inside a larger architecture that realizes a mechanism.
Mechanisms
conformational state discriminationConformational Uncagingnanopore-based single-molecule sensingTechniques
No technique tags yet.
Target processes
No target processes tagged yet.
Implementation Constraints
The reported implementation couples glass nanopipette-based single-cell extraction to silicon nitride solid-state nanopore measurements. Beyond this workflow and the use of LOV2 as the demonstrated target, the supplied evidence does not provide construct design details, buffer conditions, pore fabrication parameters, or delivery requirements.
The supplied evidence describes one reported application centered on LOV2, so generality across other proteins, cell types, and assay conditions is not established here. The evidence also indicates that measurements from direct single-cell samples differ significantly from purified samples, which highlights biological relevance but also implies that purified-sample behavior may not transfer straightforwardly to this workflow.
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Using glass nanopipette-based single-cell extraction together with SiNx nanopores, the authors identified LOV2 and monitored its conformational changes from single-cell extracts.
we performed ex situ single-cell protein analysis by directly extracting the contents of individual cells using glass nanopipette-based single-cell extraction and successfully identified and monitored the conformational changes of the LOV2 protein from single-cell extracts using SiN x nanopores
Using glass nanopipette-based single-cell extraction together with SiNx nanopores, the authors identified LOV2 and monitored its conformational changes from single-cell extracts.
we performed ex situ single-cell protein analysis by directly extracting the contents of individual cells using glass nanopipette-based single-cell extraction and successfully identified and monitored the conformational changes of the LOV2 protein from single-cell extracts using SiN x nanopores
Using glass nanopipette-based single-cell extraction together with SiNx nanopores, the authors identified LOV2 and monitored its conformational changes from single-cell extracts.
we performed ex situ single-cell protein analysis by directly extracting the contents of individual cells using glass nanopipette-based single-cell extraction and successfully identified and monitored the conformational changes of the LOV2 protein from single-cell extracts using SiN x nanopores
Using glass nanopipette-based single-cell extraction together with SiNx nanopores, the authors identified LOV2 and monitored its conformational changes from single-cell extracts.
we performed ex situ single-cell protein analysis by directly extracting the contents of individual cells using glass nanopipette-based single-cell extraction and successfully identified and monitored the conformational changes of the LOV2 protein from single-cell extracts using SiN x nanopores
Using glass nanopipette-based single-cell extraction together with SiNx nanopores, the authors identified LOV2 and monitored its conformational changes from single-cell extracts.
we performed ex situ single-cell protein analysis by directly extracting the contents of individual cells using glass nanopipette-based single-cell extraction and successfully identified and monitored the conformational changes of the LOV2 protein from single-cell extracts using SiN x nanopores
SiNx solid-state nanopores can be used for single-molecule protein analysis from complex cellular samples.
Here, we explored the potential of SiN x solid-state nanopores for single-molecule protein analysis from complex cellular samples.
SiNx solid-state nanopores can be used for single-molecule protein analysis from complex cellular samples.
Here, we explored the potential of SiN x solid-state nanopores for single-molecule protein analysis from complex cellular samples.
SiNx solid-state nanopores can be used for single-molecule protein analysis from complex cellular samples.
Here, we explored the potential of SiN x solid-state nanopores for single-molecule protein analysis from complex cellular samples.
SiNx solid-state nanopores can be used for single-molecule protein analysis from complex cellular samples.
Here, we explored the potential of SiN x solid-state nanopores for single-molecule protein analysis from complex cellular samples.
SiNx solid-state nanopores can be used for single-molecule protein analysis from complex cellular samples.
Here, we explored the potential of SiN x solid-state nanopores for single-molecule protein analysis from complex cellular samples.
Proteins measured directly from single cells differed significantly from proteins obtained from purified samples.
Our results reveal significant differences between proteins measured directly from single cells and those obtained from purified samples.
Proteins measured directly from single cells differed significantly from proteins obtained from purified samples.
Our results reveal significant differences between proteins measured directly from single cells and those obtained from purified samples.
Proteins measured directly from single cells differed significantly from proteins obtained from purified samples.
Our results reveal significant differences between proteins measured directly from single cells and those obtained from purified samples.
Proteins measured directly from single cells differed significantly from proteins obtained from purified samples.
Our results reveal significant differences between proteins measured directly from single cells and those obtained from purified samples.
Proteins measured directly from single cells differed significantly from proteins obtained from purified samples.
Our results reveal significant differences between proteins measured directly from single cells and those obtained from purified samples.
Fusing a nanopore electrophoretic driver protein to LOV2 enhanced capture efficiency of the target protein.
we designed a nanopore electrophoretic driver protein and fused it with LOV2, thereby enhancing the capture efficiency of the target protein
Fusing a nanopore electrophoretic driver protein to LOV2 enhanced capture efficiency of the target protein.
we designed a nanopore electrophoretic driver protein and fused it with LOV2, thereby enhancing the capture efficiency of the target protein
Fusing a nanopore electrophoretic driver protein to LOV2 enhanced capture efficiency of the target protein.
we designed a nanopore electrophoretic driver protein and fused it with LOV2, thereby enhancing the capture efficiency of the target protein
Fusing a nanopore electrophoretic driver protein to LOV2 enhanced capture efficiency of the target protein.
we designed a nanopore electrophoretic driver protein and fused it with LOV2, thereby enhancing the capture efficiency of the target protein
Fusing a nanopore electrophoretic driver protein to LOV2 enhanced capture efficiency of the target protein.
we designed a nanopore electrophoretic driver protein and fused it with LOV2, thereby enhancing the capture efficiency of the target protein
Approval Evidence
1 source3 linked approval claimsfirst-pass slug sinx-solid-state-nanopore-single-cell-protein-analysis-approach
Here, we explored the potential of SiN x solid-state nanopores for single-molecule protein analysis from complex cellular samples... successfully identified and monitored the conformational changes of the LOV2 protein from single-cell extracts using SiN x nanopores.
Source:
application resultsupports
Using glass nanopipette-based single-cell extraction together with SiNx nanopores, the authors identified LOV2 and monitored its conformational changes from single-cell extracts.
we performed ex situ single-cell protein analysis by directly extracting the contents of individual cells using glass nanopipette-based single-cell extraction and successfully identified and monitored the conformational changes of the LOV2 protein from single-cell extracts using SiN x nanopores
Source:
capabilitysupports
SiNx solid-state nanopores can be used for single-molecule protein analysis from complex cellular samples.
Here, we explored the potential of SiN x solid-state nanopores for single-molecule protein analysis from complex cellular samples.
Source:
comparisonsupports
Proteins measured directly from single cells differed significantly from proteins obtained from purified samples.
Our results reveal significant differences between proteins measured directly from single cells and those obtained from purified samples.
Source:
Comparisons
Source-backed strengths
The approach was demonstrated for single-molecule protein analysis from complex cellular samples using SiNx solid-state nanopores. A key validated result is direct identification of LOV2 and monitoring of its conformational changes from single-cell extracts, showing that the method can resolve protein state information in material obtained from individual cells.
Ranked Citations
- 1.