Source 1primary paper2016Scientific Reports
Toolkit/antibody adsorption single-molecule environment
antibody adsorption single-molecule environment
Also known as: antibody adsorption
Taxonomy: Technique Branch / Method. Workflows sit above the mechanism and technique branches rather than replacing them.
Summary
We demonstrated that the same fluorophore photoswitching properties measured in PVA films and using antibody adsorption, a protein-conjugation environment analogous to labeled cells, were significantly correlated to microtubule width and continuity, surrogate measures of SMLM image quality.
Usefulness & Problems
Why this is useful
Antibody adsorption provides a single-molecule measurement environment for fluorophore photoswitching. The paper uses it as a comparison condition alongside PVA films.; measuring fluorophore photoswitching properties in a protein-conjugation-like environment; comparing screening measurements against an environment analogous to labeled cells
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Antibody adsorption provides a single-molecule measurement environment for fluorophore photoswitching. The paper uses it as a comparison condition alongside PVA films.
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measuring fluorophore photoswitching properties in a protein-conjugation-like environment
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comparing screening measurements against an environment analogous to labeled cells
Problem solved
It offers a comparison context that is closer to labeled-cell conditions than PVA alone. This helps relate measured photoswitching properties to imaging outcomes.; provides a comparison environment analogous to labeled cells for fluorophore photoswitching assessment
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It offers a comparison context that is closer to labeled-cell conditions than PVA alone. This helps relate measured photoswitching properties to imaging outcomes.
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provides a comparison environment analogous to labeled cells for fluorophore photoswitching assessment
Problem links
provides a comparison environment analogous to labeled cells for fluorophore photoswitching assessment
LiteratureIt offers a comparison context that is closer to labeled-cell conditions than PVA alone. This helps relate measured photoswitching properties to imaging outcomes.
Source:
It offers a comparison context that is closer to labeled-cell conditions than PVA alone. This helps relate measured photoswitching properties to imaging outcomes.
Published Workflows
Objective: Establish a quantitative screening methodology for fluorophore photoswitching that predicts SMLM image quality without requiring protein conjugation before assessment.
Why it works: The workflow is expected to work because photoswitching properties measured in simplified single-molecule environments were reported to correlate significantly with surrogate measures of SMLM image quality.
fluorophore photoswitching behavior linked to localization microscopy image formationsingle-molecule environment-based screeningquantitative photoswitching characterizationcorrelation of screening measurements with image-quality surrogates
Stages
- 1.Quantify fluorophore photoswitching in PVA films(broad_screen)
This stage exists to provide a facile single-molecule screening environment for systematic fluorophore characterization without requiring protein conjugation first.
Selection: Efficient quantification of fluorophore photoswitching properties in a single-molecule environment
- 2.Measure photoswitching using antibody adsorption comparison environment(secondary_characterization)
This stage exists to compare PVA-based measurements against an environment analogous to labeled cells.
Selection: Assess the same photoswitching properties in a protein-conjugation environment analogous to labeled cells
- 3.Correlate measured photoswitching properties with SMLM image-quality surrogates(confirmatory_validation)
This stage exists to test whether measured photoswitching properties predict image-quality outcomes relevant to SMLM.
Selection: Significant correlation to microtubule width and continuity as surrogate measures of SMLM image quality
Taxonomy & Function
Primary hierarchy
Technique Branch
Method: A concrete measurement method used to characterize an engineered system.
Techniques
Functional AssayTarget processes
No target processes tagged yet.
Input: Light
Implementation Constraints
cofactor dependency: cofactor requirement unknownencoding mode: genetically encodedimplementation constraint: context specific validationimplementation constraint: spectral hardware requirementoperating role: sensor
The approach requires antibody adsorption as the measurement environment. The abstract frames it as a protein-conjugation environment analogous to labeled cells.; requires antibody adsorption setup; used as a comparison environment for photoswitching measurements
The abstract does not present antibody adsorption as removing the protein-conjugation hurdle. Instead, that hurdle is part of the motivation for validating PVA.; still described as a protein-conjugation environment
Validation
Cell-freeBacteriaMammalianMouseHumanTherapeuticIndep. Replication
Supporting Sources
Ranked Claims
Polyvinyl alcohol (PVA) was validated as a single-molecule environment to efficiently quantify fluorophore photoswitching properties.
Herein, we validated polyvinyl alcohol (PVA) as a single-molecule environment to efficiently quantify the photoswitching properties of fluorophores
Fluorophore photoswitching properties measured in PVA films and by antibody adsorption were significantly correlated with microtubule width and continuity, surrogate measures of SMLM image quality.
We demonstrated that the same fluorophore photoswitching properties measured in PVA films and using antibody adsorption, a protein-conjugation environment analogous to labeled cells, were significantly correlated to microtubule width and continuity, surrogate measures of SMLM image quality.
Defining PVA as a fluorophore photoswitching screening platform will facilitate SMLM fluorophore development and optimal image buffer assessment through facile and accurate photoswitching property characterization that translates to imaging performance.
Defining PVA as a fluorophore photoswitching screening platform will facilitate SMLM fluorophore development and optimal image buffer assessment through facile and accurate photoswitching property characterization, which translates to SMLM fluorophore imaging performance.
Approval Evidence
1 source1 linked approval claimfirst-pass slug antibody-adsorption-single-molecule-environment
We demonstrated that the same fluorophore photoswitching properties measured in PVA films and using antibody adsorption, a protein-conjugation environment analogous to labeled cells, were significantly correlated to microtubule width and continuity, surrogate measures of SMLM image quality.
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predictive associationsupports
Fluorophore photoswitching properties measured in PVA films and by antibody adsorption were significantly correlated with microtubule width and continuity, surrogate measures of SMLM image quality.
We demonstrated that the same fluorophore photoswitching properties measured in PVA films and using antibody adsorption, a protein-conjugation environment analogous to labeled cells, were significantly correlated to microtubule width and continuity, surrogate measures of SMLM image quality.
Source:
Comparisons
Source-stated alternatives
The main alternative named in the abstract is PVA films, which are proposed as a more facile screening platform. The abstract also references conventional protein-conjugation-based assessment more broadly.
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The main alternative named in the abstract is PVA films, which are proposed as a more facile screening platform. The abstract also references conventional protein-conjugation-based assessment more broadly.
Source-backed strengths
analogous to labeled cells
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analogous to labeled cells
Compared with polymer films
The main alternative named in the abstract is PVA films, which are proposed as a more facile screening platform. The abstract also references conventional protein-conjugation-based assessment more broadly.
Shared frame: source-stated alternative in extracted literature
Strengths here: analogous to labeled cells.
Relative tradeoffs: still described as a protein-conjugation environment.
Source:
The main alternative named in the abstract is PVA films, which are proposed as a more facile screening platform. The abstract also references conventional protein-conjugation-based assessment more broadly.
The main alternative named in the abstract is PVA films, which are proposed as a more facile screening platform. The abstract also references conventional protein-conjugation-based assessment more broadly.
Shared frame: source-stated alternative in extracted literature
Strengths here: analogous to labeled cells.
Relative tradeoffs: still described as a protein-conjugation environment.
Source:
The main alternative named in the abstract is PVA films, which are proposed as a more facile screening platform. The abstract also references conventional protein-conjugation-based assessment more broadly.
Ranked Citations
- 1.